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Cell therapy / cellular therapy A recent revision of national application documents (SF INF 50) added a new field (human biology) and subdomain (human derived products) to include analytical methods perform to characterize therapeutic cellular products. One requirement of AB001 ISO 15189 standards is to validate analytical methods. For Long term follow up after granulocyte transfusions methods that use a diagnostic and follow manufacturer C. Kreissig1,*, I. Fischer1 instructions, a verification shall be performed. This verification 1ZBST, DRK-BLUTSPENDEDIENST WEST, Ratingen, Germany is detailed in french guidelines (SH GTA 04 et SH GTA 14). This abstract report our verification results for CD34 þ cell Introduction: Indication for granulocyte transfusion is put numeration with Stem-kit reagent, performed on FC500 flow seldom, because collection of products is complex, time cytometer. consuming and costly. That’s why only very ill patients get Materials (or patients) and methods: Intra assay precision granulocyte transfusions. was measured with peripheral blood and fresh or frozen/ We wanted to determine, if big effort is justified not only by thawed cellular products samples, quantifying the difference the fact, that patient survive acute infection, but also by long between duplicate samples. We analyzed separately term survival and good health. samples with a concentration lower than 99 CD34/microL Materials (or patients) and methods: We performed a retro- (level 1) and those upper than 100 CD34/microL (level 2). spective analysis of one year follow up after granulocyte transfusions. Intermediate precision, precision between laboratories, Via a questionnaire we determined patient status at four time accuracy and quantification limit were measured with points: when stopping transfusions, 4 weeks, 6 month and one stabilized blood used for QC (Status Flow Pro, Eurocell year after last granulocyte transfusion. diagnostics). Two levels are available (11 and 32 CD34/microL). In this abstract we correlate one year survival with age, underlying Finally, methods comparison consisted in an analysis disease, indication for granulocyte transfusion, number of of the same sample measured with Stem-kit reagents granulocyte transfusions and dose of transfused granulocytes. but analyzed either on FC500 (Beckman Coulter) or FACS Results: We treated 28 patients, 11 female, 17 male. They were 2 Calibur (Becton Dickinson) flow cytometers. Samples to 69 years old, 17 patients were pediatric, 11 adult. Underlying compared were peripheral blood and fresh or frozen/ thawed disease was in 17 patients leukemia, in 7 patients inborn diseases, cellular products, with a concentration range from 1.5 to 159 in 2 patients lymphoma, in one patient solid tumor, in one CD34/microL. patient unwanted side effect of Novamin taking. Reasons for Results: Performance, accuracy and methods comparison are granulocyte transfusion were in 12 patients localized infection, in reported in the following table: 10 patient septicemia and in 6 patients fever of unknown origin. Number of transfusions was 1 till 23. Dose of transfused granulocyte ranged from 0,4 till 5,5x10E9 per kg bodyweight. Intra Intermediate Precision Accuracy Methods 7 out of 28 patients died during granulocyte transfusion series. assay precision between comparison In 5 of 21 surviving patients follow up data were not available. precision laboratories One year after granulocyte transfusions 4 patients were alive and well. nCV n CV (%) n Biais %n r2 Surviving patients were 2, 10, 16 and 20 years old. They suffered (%) (%) from leukemia (2 patients) or inborn disease (2 patients). Reason for granulocyte transfusion was septicemia (1 patient), level 1 32 14,2 15 10,8 15 2,2 76 36 0,988 fever of unknown origin (1 patient) and localized infection (2 level 2 20 5,1 18 5,4 18 7,8 84 patients). Patients were given 3 till 23 transfusions, doses ranged from 0,7 till 4,6 x10E9 granulocytes per kg bodyweight. Conclusion: 4 out of 16 patients (25%) were alive and well one Accuracy interpolation was 75% at a concentration of 20 year after granulocyte transfusion. So we conclude, granulo- CD34/microL. Measured quantification limit was 3 CD34/ cyte transfusion is a useful treatment opportunity for seriously microL. ill patients. Conclusion: These measures allowed us to verify our Wecouldnotfindanycorrelation of one year survival with age, CD34 þ cell quantification method was suitable for underlying disease, indication for granulocyte transfusion, number samples and concentrations we use. We are confident for of granulocyte transfusions and dose of transfused granulocytes. clinical decisions that are taken on these results. Accuracy Disclosure of Interest: None declared. has to be taken in consideration in some circumstances. If institutional SOP is to start an apheresis when peripheral blood CD34 þ cell concentration is upper than 20/microL, AB002 knowing our accuracy at this concentration (75%) Method verification for CD34 þ cell numeration with we can decide to start the apheresis as soon as blood Stem-kit reagents according to ISO15189 standards concentration reaches 17 CD34/microL. CD34 þ cell C. Lemarie1,2,*, J. Gaude1, B. Makni1, B. Calmels1,2, M. Bouyssie3, concentration of some thawed cord bloods is under quanti- C. Chabannon1,2 fication limit. Clinicians shall be informed of the lack of 1Centre de The´rapie Cellulaire, De´partement de Biologie du Cancer, accuracy of these low results. Finally, national accreditation Institut Paoli-Calmettes, 2Centre d’Investigations Cliniques en committees have to confirm analytical methods performed to Biothe´rapie, CBT-1409, 3Service qualite´ et gestion des risques, qualify cellular products are in the field of ISO 15189 Direction ge´ne´rale, Institut Paoli-Calmettes, Marseille, France standards. Disclosure of Interest: None declared. Introduction: French biological laboratories shall be accredi- tated by COFRAC comitee, according to ISO15189 standards.

S553 AB003 than 1% T cells. Sterility testing was negative for bacteria, Untouched GMP-grade purified engineered immune cells funghi and yeast,o10 CFU Mycoplasma/ml and o1.6 IU/ml to treat cancer endotoxin tested according to the European Pharmacopeia. T. Straetemans1, C. Grunder1, S. [email protected], Results: The overall production data are shown in the table S. Hol1, I. Slaper-Cortenbach2, H. Bonig3, Z. Sebestye´n1, J. Kuball1,* below. From the 47 batches produced so far, one batch was 1Hematology/Laboratory of Translational Immunology, 2Cell not released due to cell clumping. Therapy Facility, University Medical Center Utrecht, Utrecht, There was no difference in the expansion of fresh BM or Netherlands, 3Institute for Transfusion Medicine and Immunohe- cryopreserved BM. It was also shown that MSCs expanded matology, Johann-Wolfgang-Goethe University, Frankfurt, from younger donors resulted in a better yield of MSC. Germany Cryopreserved P2 cells showed excellent reproducibility in their expansion towards P3, not only in the expansion period, Introduction: Engineering T cells with receptors to re-direct but also in the yield of MSCs per run. We did not detect a the immune system against cancer has most recently been difference in the PL batches. After thawing of the bags and the described as one of the scientific breakthroughs. However, no reference vials (n ¼ 46), a correlation coefficient of 0.935 was additional purification step for engineered immune cells is measured for viable MSCs in bags and vials. being applied due to the lack of suitable tools and strategies. This results in the transfer of also non- and poorly engineered immune cells into patients, which can substantially dampen Passage Yield cells x 106/ 2CS (min-max) Day therapeutic effects and limit additional clinical applications such as the transfer of third party populations. In the present 1 54,8 (3,83–117,5) 10 study we therefore developed a cost-effective GMP-compliant 2 49,0 (9,0–115,8) 6 system to obtain highly purified engineered immune cells. 3 59,1 (7,5–115,6) 6 Materials (or patients) and methods: By taking advantage of an optimized expression system, a model receptor naturally interfering with endogenous abTCR chains, and GMP-grade Conclusion: The production of MSC in 2 layer CS resulted in anti-abTCR beads we propose a novel strategy to isolate high numbers of MSC available for clinical application. The engineered immune cells. production of MSC in 2CS using PL pools is highly reproducible Results: The untouched enrichment of engineered immune and a good correlation was seen between the reference vials cells translated into highly purified receptor engineered cells and the bags. with strong anti-tumor reactivity both in vitro but also in vivo Disclosure of Interest: None declared. in two different humanized mouse models. Importantly, this approach also eliminated residual allo-reactivity of engineered AB005 immune cells. Our data demonstrate that even with long-term Efficacy and side effects of granulocyte transfusion suboptimal interference with endogenous TCR chains such as therapy in eight hematological patients in resting cells, allo-reactivity remained absent and tumor L. Minculescu1,*, E. Haastrup1, M. Ifversen2, A. Fischer-Nielsen1 control preserved. 1 2 Conclusion: All together, we present a novel GMP-grade Dept of Clinical Immunology, Department of Pediatrics, enrichment method of untouched engineered immune cells, Rigshospitlaet, Copenhagen, Denmark which is potentially applicable to all receptor-modified cells even if interference with endogenous TCR chains is far from Introduction: There are currently no definite guidelines for complete granulocyte transfusion therapy (GTX) for long-term neutro- Disclosure of Interest: None declared. penia in patients with severe infections unresolved by conventional anti-infectious treatment (1,2). Here we describe our retrospective, observational study of GTX in eight Danish AB004 patients treated between 2007 and 2014. Production results of Mesenchymal Stromal Cells cultured Materials (or patients) and methods: Indication for GTX was in CellSTACKs and platelet-lysate for clinical application neutropenia (o0,5x109/L) and severe infection unresolved by K. Westinga1,*, M. van Gelder1, S. Vonk-Griffioen1, P. Leeflang1, conventional anti-infectious treatment. Granulocytes were N. Wulffraat2, J. Kuball3, M. van den Broek1, I. Slaper-Cortenbach1 collected from related and unrelated ABO group-matched 1Dept of Clinical Pharmacy, Cell Therapy Facility, 2Department of donors. For neutrophil mobilization, donors received one Paediatrics / Immunology,Rheumatology and Infectiology, subcutaneous dose of 0.3 mg non-glycosylated G-CSF (Filgras- 3Hematology/Laboratory of Translational Immunology, UMC tim) and Prednisolone 50 mg 10-12 hours prior to apheresis. Utrecht, Utrecht, Netherlands Collection processes were performed on COBEs Spectra or Spectra Optias Apheresis System from Terumo BCT. Introduction: In our GMP-accredited facility, we have Results: Transfused granulocytes contained a median neu- manufactured Mesenchymal Stromal Cells (MSCs) for several trophil number of 2.4x1010 (range 0.4-5.2x1010) per unit, a clinical trials using platelet lysate (PL) as the sole source of median of 3.3x108 neutrophils per kg bodyweight. Patients growth factors. The aim of this study is to evaluate the received a median of 5 (range 1-21) granulocyte transfusions. production results of 47 batches and the reproducibility of the Patient outcomes are summarized in table 1. Infections production data. resolved in 2 out of 8 patients, where a decrease in C-reactive Materials (or patients) and methods: MSC were isolated from protein levels together with a haematological response in bone marrow from 10 male and 9 female healthy volunteers terms of a sustained increase in neutrophil count40.5x109/L and isolated and expanded using 2-layered CellSTACK (2CS) was observed shortly after initiation of GTX treatment. One culture systems in combination with Macopharma tubing sets/ patient deteriorated rapidly as a result of respiratory failure in bags for seeding, washing and harvesting. Mononuclear cells direct association to granulocyte infusion. This patient had (200 106/CS) were cultured in a-MEM, 5% PL and 1000 IU impaired lung function as a result of severe fungal infection Heparin/2CS. MSCs were passaged at full confluency using prior to GTX. Chest X-ray showed bilateral interstitial infiltrates TrypLE. Per 2CS 2–5 106 MSC were seeded for expansion. compatible with transfusion-related-acute-lung-injury (TRALI), Intermediate products (P1 and P2 MSC) were cryopreserved and the patient died two weeks later from respiratory failure. and stored till further processing. For clinical application, P3 No other patients experienced severe adverse events. The MSCs were cryopreserved in 20 ml bags, containing 20 up to remaining 5 patients experienced sporadic peaks of increase 100 106 MSC per bag in Physiological Salt Solution, 5% in granulocyte concentrations (40.5x109/L) but no sustained Human Serum Albumin and 10% DMSO. MSC were 495% increase was obtained, and infections remained unresolved positive for CD 73, CD90 and CD105 McAbs and contained less resulting in death.

S554 [AB005]

Conclusion: Patients received GTX with a satisfactory median by developing a semi-automated processing protocol based neutrophil number42x1010 per unit as recommended (1,2). on our MarrowXpresss (‘‘MXP’’) technology which comprises a Decrease in C-reactive protein together with sustained functionally closed system for rapid BM processing with increase in neutrophil during GTX was observed in 2 surviving minimal manipulation and without drug additives. patients. We experienced one case interpreted as TRALI in a Materials (or patients) and methods: The MXP technology is patient with pre-existing severe fungal lung infection. Larger under optimization to yield low-hematocrit products from BM and preferably randomized patient trials are needed in order samples. Samples will be subjected to density stratification by to determine the safety and efficacy of this treatment. centrifugation enabling segregation of RBCs, white blood cells References: (1) Massey E, Paulus U, Doree C, Stanworth S. and plasma, yielding a concentrated product with high Granulocyte transfusions for preventing infections in patients recoveries of nucleated cells. Importantly, MXP processing will with neutropenia or neutrophil dysfunction. Cochrane output an autologous minimally manipulated cell concentrate Database Syst Rev 2009;(1):CD005341. sample that does not require the use of density gradient (2) Strauss RG. Role of granulocyte/neutrophil transfusions for media such as HES or Ficoll. MXP devices are semi-automated haematology/oncology patients in the modern era. Br J with internal monitoring and controls assuring repeatability. Haematol 2012 August;158(3):299-306. Furthermore, BM processing is conducted in a functionally Disclosure of Interest: None declared. closed system minimizing the probability of product contam- ination. The MXP technology is expected to be suitable for the treatment of a variety of indications including Severe AB006 Combined Immunodeficiency, aplastic anemia, thalassemia, MarrowXpresst (‘‘MXP’’) – A Functionally Closed, Semi- sickle cell anemia, and hematological malignancies. Automated, System Being Developed for Ficoll-Free Bone Results: Given the importance of minimizing the RBC content Marrow Processing for Transplantation of Concentrated in HCT products, the MXP technology is well positioned to Low-Hematocrit Products produce cell concentrates with low hematocrit and high cell C. Kim1, M. D. Lacher2,*, P. Clift1, S. Wilcox1, J. Heiserman2, recovery. Such performance enhancements are achievable by D. Sethi2, M. Sivilotti2, V. Ponemone3,4, H. Al-Esawi 1, K. Harris2 optimization of optical detection, new algorithms targeting 1Cesca Therapeutics, Inc., Rancho Cordova, 2Cesca Therapeutics, fluid flow dynamics, and centrifugation protocols. The MXP Inc., Emeryville, United States, 3Cesca Therapeutics, Inc., technology described here demonstrates processing of 4Fortis-TotipotentRX Centre for Cellular Medicine, Gurgaon, India peripheral blood to produce a white blood cell concentrate with a hematocrit less than or equal to 2% (B0.4 mL of packed Introduction: ABO blood type mismatch in hematopoietic cell RBCs per 21 mL of product volume) with cell recoveries at least transplantation (HCT) procedures may lead to serious adverse equivalent to those obtained via standard Ficoll-based effects, including GvHD. In particular, transplantation of donor procedures (Status: 15DEC2014). red blood cells (RBCs) expressing A and/or B antigens can Conclusion: The objective for this approach is to clinically trigger anti-A and/or -B antibody production in blood type O manage ABO-incompatible allogeneic HCT, with the goal of recipients resulting in hemolysis of donor RBCs. Conversely, reducing transplant-related morbidity and mortality. Further serum (or ‘‘passenger lymphocytes’’) from A, B, or O donors improvements in the MXP technology are expected to yield may contain anti-B, anti-A, or both types of antibodies leading reliable Ficoll-free low-RBC protocols. to hemolysis in the recipient. Thus, minimizing the RBC Disclosure of Interest: C. Kim Employee of: Cesca Therapeu- content in donor bone marrow (BM) is important in order to tics, Inc., M. Lacher Employee of: Cesca Therapeutics, Inc., P. minimize the risk of adverse events. We will address this issue Clift Employee of: Cesca Therapeutics, Inc., S. Wilcox Employee

S555 of: Cesca Therapeutics, Inc., J. Heiserman Employee of: Cesca acute leukemia patients after allogeneic hematopoietic stem Therapeutics, Inc., D. Sethi Employee of: Cesca Therapeutics, cell transplantation. Transfusion 2014; 54:1493-1500 Inc., M. Sivilotti Employee of: Cesca Therapeutics, Inc., V. Disclosure of Interest: None declared. Ponemone Employee of: Cesca Therapeutics, Inc., H. Al-Esawi Employee of: Cesca Therapeutics, Inc., K. Harris Employee of: Cesca Therapeutics, Inc. Hematopoietic stem cells AB007 Donor Lymphocyte Infusion – 7-years experience S. Gouveia1,*, M. J. Gutierrez2, E. Pereira2, G. Teixeira2, I. Sousa Ferreira2, N. Miranda2, M. Abecasis2 AB008 1Immunohemotherapy Department, Hospital Curry Cabral - Maximum storage time that affect the viability of stem CHLC, EPE, 2Bone Marrow Transplant Unit, Heamatology cells Department, Instituto Portugueˆs de Oncologia de Lisboa S. solmaz medeni1, C. acar1,*, S. yigitkaya1, D. turkyilmaz1, Francisco Gentil, Lisbon, Portugal G. sevindik1, I. Alacacioglu1, F. demirkan1, F. yuksel1, M. A. ozcan1, O. piskin1, B. undar1, G. H. ozsan1 Introduction: Relapse of hematological malignancies following 1hematology, DOKUZ EYLUL UNIVERSITESI TIP FAKULTESI,˙ Izmir, allogeneic hematopoietic stem cells transplantation (allo-HSCT) Turkey remains a major cause of treatment failure. Treatment options include withdrawal of immunossupression, , Introduction: Cryopreservation is needed for hematopoietic donor lymphocyte infusions (DLIs) or second allo-HSCT. stem cell viability and functional integrity when they are Through its graft-versus-leukemia effect DLI are an effective dissolved and for infusion with minimum toxicity. Dimethyl strategy. However, response to DLIs is dependent upon the sulfoxide (DMSO) and hydroxy etilen starch (HES) are used for type of disease, cell dose and development of graft vs. host this purpose. To evaluate viability of stem cells, trypan disease (GvHD). The best results are seen in patients with methylen blue staining under microscopy or flowcyto- chronic myeloid leukemia (CML) followed by patients with metry are used. In the literature, stem cells can be preserved lymphomas, multiple myeloma (MM) and acute leukemias. up to 5 years at -80 C. We want to evaluate stem cell viability Materials (or patients) and methods: Retrospective review of at 3. and 5. year after thawing, relationship between CD34 þ all patients (pts) given DLI after HSCT relapse at our institution, cell level, stem cell viability and storage time and intend to in the past 7 years. contribute to the literature Between January 2007 and December 2013, 14 pts, aged 18 to Materials (or patients) and methods: Stem cells product of 64 years (average 38.4 years) were given 22 DLIs to treat 24 patients who are ex at study time were evaluated. relapse of a myeloid (7 pts) or lymphoid malignancy (7 pts). Demograhic features, CD34 þ cell levels at the time it was Nine pts were transplanted with a related donor and 5 with an collected and after dissolution with trypan blue, flow unrelated one. cytometry in 7AAD CD 34 viability test and storage time were Results: The median time between allo-HSCT and DLI was 290 analyzed. days (68–2062). Results: Median age was 48.6 (19-65) with M/F ratio as 70,8/ 5 pts had GVHD before DLI infusion, of whom 3 died of 29,2. Mean CD34 þ cell level at product at the collected time progressive disease. was 1688 ml (120-7457), but 292 ml (4-1345) after disolved. Most patients (10–71.4%) received only one DLI. 2 pts (14.3%) Viability was 33.9% (1-78) when they are disolved. Mean received two infusions and other 2 pts (14.3%) three infusions. product storage period was 59 months (35-135). There was a The administered CD3 þ dosages were: inverse correlation between storage time and CD34 þ cell level (at beginning and at the end of storage time). Viability was lower at long-term stored product. The patient were grouped according to stored time of their product as o48 Dosage (CD3 þ /Kg) months (n:10), 48-60 months (n:3), 460 months (n:11). CD34 þ cell viability was higher in o48 months with respect Related donor N Unrelated donor N to 460 months. The most lower viability was seen at 460 months group but this was not statistically significant. CD34 st þ 1 infusion 5x10^6 (4.88x10^6–1.14x10^7) 9 1.13x10^6 5 cell level also very lower at 460 months group without (1x10^6–5x10^7) 2nd infusion 1.08x10^7 (1x10^7–1.04x10^8) 4 5x10^6 2 statistical significance. The lack of statistical significance 3rd infusion 2x10^7 1 1x10^7 1 attributed the small number of patients. Conclusion: In conclusion; viability of stem cells and amount of CD34 þ cells in the product are affected by storage time 12 pts had no response, with 8 dying from progressive disease. period and especially storage time longer than 60 months 2 patients, both with ALL, have successfully responded to the mostly affect the viability of stem cell in the product detected DLI, and are currently in complete remission. with both trypan blue staining and flowcytometric analysis. Conclusion: Although this is a small and heterogeneous Disclosure of Interest: None declared. series, we note that, at our institution, most patients treated with DLI receive only one dose. The occurrence of GvHD before or after DLI did not predict for AB009 favorable therapeutic response. Haploidentical Hematopoietic Stem Cell Transplant Patients with ALL had the better outcome (100% response). For Sickle Cell Disease Using Thiotepa Based Reduced References: Intensity Conditioning And Post Transplant 1. Deol A., Lum L.G. Role of donor lymphocyte infusions in Cyclophosphamide relapsed hematological malignancies after stem cell trans- D. G. Kharya1,*, D. Doval1, D. R. Chaudhary1, M. Dhamija1, plantation revisited. Cancer Treatment Reviews 36 (2010) V. Khandelwal1, S. Lunkad1, R. Setia1, A. Handoo1, S. Sharma1, 528–538 T. Dadu1 2. Chang Y. J., Huang X.-J. Donor lymphocyte infusions for 1BLK Super Speciality Hospital, Delhi, India, Delhi, India relapse after allogeneic transplantation. When, if and for whom? Blood Reviews 27 (2013) 55–62 Introduction: Hematopoietic stem cell transplant (HSCT) can 3. Tan Y., Du K., Luo Y. et al. Superiority of preemptive donor cure sickle cell disease (SCD). This option is however limited by lymphocyte infusion based on minimal residual disease in availability of matched sibling or related unaffected donors.

S556 Over the years results of haploidentical transplants have Results: Engraftment was observed in all pts with respect to improved significantly. Bolanos et al reported successful count of polymorphonuclears after 12 (11-15) days and haploidentical HSCT for SCD using post transplant cyclopho- platlets after 11 (9-17) days. Acute (a) GvHD was observed in sphamide (PTCy) however there cohort had almost 40% risk of 9 pts (56,25%), advanced grade III/IV in 2 pts (12,5%) and rejection. We report a child with SCD with complications being chronic (c) GvHD in 12 pts (75%). Overall survival (OS) of all our successfully treated using thiotepa based reduced intensity pts is 37,5% with median follow up 42 (24-204) months. Vast conditioning along with PTCy. majority of alive pts had mild or moderate cGvHD without Materials (or patients) and methods: 3 year old boy with serious influence on their quality of life. Causes of death in 10 SCD on regular transfusions was taken up for haploidentical (62,5%) pts were relapses in 4 (25%) pts after 6, 12, 24 and 29 HSCT from father in the absence of suitably matched donor. months after secondary SCT, 2 (12,5%) pts have died from He was started on hypertransfusion (target Hb 11-13gm/dl) VOD, 1 (6,25%) pt due to infection and 2 (12,5%) pts had and hydroxyurea (20 mg/kg) from day -45. He was conditioned severe cGvHD. TRM was 6,25%. using Thiotepa 10 mg/kg in two divided doses (D-7), Conclusion: Our modest results have showed benefit of Fludarabine 30 mg/m2 (D-6 to D-2), Cyclophosphamide second SCT as treatment option for selected cohort of pts who 14.5 mg/kg (D-5, D-4), TBI 2 Gy with thymic shielding (D-1), have failed after first allografting suggesting that relapses still Thymoglobulin 1.5 mg/kg (D-9 to D-7). GVHD prophylaxis remains major cause of failure after allogeneic SCT. included PTCy 50 mg/kg/day on D3 and 4, Tacrolimus to Disclosure of Interest: None declared. maintain a level of 5-15 ng/ml (till 6 months post HSCT) & MMF (till D35) starting from D5. Patient received unmanipulated AB011 GCSF primed bone marrow harvested from his father with a 8 Adverse Reactions Occurring in The Patients Within One target MNC 48x10 /kg. He received 9.03x10^8/kg mono- Hour Following Hematopoietıc Stem Cell Infusion nuclear cells, 8.41x10^6/kg CD34 cells. 1,* 1 2 1 Results: Polymorphonuclear cell and platelet engraftment N. Baskan , D. Cekdemir , E. Birtas Atesoglu , Z. Gulbas 1Hematology, Anadolu Medical Center Hospital, Bone Marrow were seen on D þ 12 and D þ 15 respectively. Whole blood 2 chimerism on day þ 15 showed 99.45% donor cells. There is Transplantation Unite, Hematology, Kocaeli University, no evidence of acute or chronic GVHD. He was discharged on Department of Internal Medicine,, Kocaeli, Turkey D þ 20 in good clinical condition. He is currently D þ 80 post HSCT clinically well, on tacrolimus. Post HSCT his sickle Introduction: Hematopoietic stem-cell transplantation (HSCT) percentage has gone down from 88% to 22%, chimerism is a therapeutic modality widely used in treatment of D þ 60 post HSCT is 99.62%. hematological diseases. In this context, undesired reactions Conclusion: Successful Haploidentical Paternal HSCT for SCD occurring in the patient during infusion of the frozen samples using thiotepa based conditioning and PTCy. This appears to is clinically important. DMSO is known to be the most be a promising technique in haploidentical or MMUD HSCT important reason for the unwanted reactions observed during with early and sustained engraftment and less risk of GVHD. infusion. Side effects of this commonly used preservative This is the first reported case of haploidentical HSCT using this agent are nausea, vomiting, diarrhea, headaches, flushing, technique of sickle cell disease in India. fever, chills, dyspnea, hypotension, hypertension, chest pain, Disclosure of Interest: None declared. anaphylaxis, vasodilation, and hepatic and abdominal com- plaints. Nausea and vomiting are the most commonly encountered side effects. We investigated the adverse AB010 reactions (AR) occurring within one hour following stem-cell Second allogeneic hematopoietic stem cell transplantation infusion in the patients receiving HSCT. as a treatment option for relapses after first Materials (or patients) and methods: A total of 219 patients transplantation- our experience including 76 (34.7%) patients with multiple myeloma (MM), 28 M. Elez1,*, D. Stamatovic1, O. Tarabar1, S. Marjanovic1, B. Balint2, (12.8%) patients with Hodgkin’s lymphoma (HL), 20 patients B. Todoric Zivanovic3, V. Skuletic3, A. Zivanovic Ivic1, O. Tasic with myelodysplastic syndrome (MDS), 13 (5.9%) patients with Radic3, M. Malesevic1, L. Tukic1 acute lymphocytic leukemia, 23 (10.5%) patients with acute 1Clinic of hematology, 2Institute for transfuziology, 3Institute for myeloid leukemia (AML), 42 (19.2%) patients with non- pathology, Military Medical Academy, Belgrade, Serbia Hodgkin lymphoma, 2 (0.9%) patients with solid tumors, 8 (3.7%) patients with chronic myeloid leukemia, 2 (0.9%) Introduction: Allogeneic stem cell transplantation (SCT) is patients with with chronic lymphocytic leukemia (CLL), 3 only potentially curative option for hematological malignan- (1.4%) patients with IM, and 2 (0.9%) patients with AA who cies, but relapses are still possible and till now there is no underwent transplantation in the Bone Marrow Transplanta- standard approach neither for relapses, nor for other SCT tion (BMT) Unit of Anadolu Medical Center Hospital between failures. Current options for treatment of relapses after first 2013 and 2014 were enrolled in this study. The patients SCT are chemotherapy, donor lymphocyte infusion or second underwent autologous, allogeneic, haploidentical, or unrelated SCT. donor transplantation. Mean age of the patients was 47 (range: Goal of this retrospective study is to estimate efficacy of 14–80) years with 82 (37.4%) of them being female and 137 second allogenic SCT in the treatment of relapses after primary (62.6%) of them being male. As premedication, the patients SCT. received 1 vial of Avil, 1 vial of Decort, 1 vial of Kytril, 1 vial of Materials (or patients) and methods: From 1995. till 2014., Novalgin in 100 cc of saline solution 1 hour before the infusion. 16 patients (pts) undergone second SCT for treatment of Source of stem cells was bone marrow in 9 patients and leukemia relapse. Median age in this cohort of pts was peripheral blood for 210 patients. One hundred and ninety 29 (12-51) years, M/F ratio 8/8. Pts were suffering from various four (88.6%) patients underwent infusion of frozen samples diseases: 4 AML, 10 ALL, 2 CML. Relapse had occured at a (FS) using DMSO while 25 (11.4%) patients underwent infusion median of 19,1 months after first allo SCT (range 7-73). All pts of unfrozen samples (UFS). Data were analyzed for percentage were conditioned with reduced intensity conditioning distribution using 21.0 version SPSS software. List of URs were (Flu þ Mel) and received stem cells originated from peripheral created and completed by preparing standard form for the blood from same identical sibling donor in 14 pts and in 2 pts patients. from other identical sibling donor. Prevention of graft versus Results: During and one hour after HSC infusion, 104 (47.5%) host disease (GvHD) was modified according to specific patients developed AR and 115 (52.5%) patients did not. Of situation (complete absence of prophylaxis in the cases of 104 patients developing AR, 96 (92.3%) received infusion of active disease in 11 pts or combination of Cyclosporin A with FSs and remaining 8 (7.7%) received infusion of UFSs. Rate of Metothrexate or MMF in 5 pts in CR2). nausea (18.6%), hypertension (Z140/90 mmHg) (12.4%),

S557 vomiting (12.4%) were found to be higher than those basal medium. Then, keratinocytes were overlaid on the receiving infusion of UFSs. hADMSC and cultured for other 7 days in complete Conclusion: Considering all evalutions, the fact that rate of AR keratinocyte medium. is significantly higher in the infusions given using FSs confirms Results: We have evaluated the growth behavior of hADMSC the hypothesis that the preservative agent DMSO plays and keratinocytes as a co-culture on the Col-HA layer. The important role in development of AR. obtained results revealed that the designed co-culture has a Disclosure of Interest: None declared. high potential for human keratinocyte proliferation. Keratino- cytes without ADMSC-support formed only an irregular layer. This suggests that mesenchymal intercellular communications are necessary for proliferation and stratification of human keratinocytes. Keratinocytes cultured with hADMSC expressed Regenerative medicine Ck10 and Ck14 in supra-basal layer. Conclusion: However ADMSC is not a part of normal skin but it could promote the epidermal regeneration as describes AB012 previously for the bone marrow stem cell and other progenitor Human Adipocyte Derived Mesenchymal Stem cell cells. Our data propose ADMSC as an efficient and safe source supports keratinocyte growth in a modified Collagen- of feeder cells for the generation of keratinocyte autografts Hyalorunic acid matrix without immunological complications. 1,* 2 1 3 References: Aoki S, Toda S, Ando T, Sugihara H. Bone marrow A. Shoae-Hassani , A. A. Hamidieh , R. Mohseni , P. keyhanvar , stromal cells, preadipocytes, and dermal fibroblasts promote A. Azimi4, S. A. Mortazavi Tabatabaei1, M. Tondar5, 6 epidermal regeneration in their distinctive fashions. Mol Biol P. Ranjbarvan Cell 2004; 15:4647-4657. 1Applied Cell Sciences, 2Hematology-Oncology Research center, 3 Kataoka K, Medina RJ, Kageyama T, Miyazaki M, Yoshino T, Tehran University of Medical Sciences, Medical Nanotechnology, Makino T, Huh NH. Participation of adult mouse bone marrow Iran University of Medical Sciences, Tehran, Iran, Islamic Republic 4 cells in reconstitution of skin. Am J Pathol 2003; 163:1227-1231. Of, Oncology-Pathology, Karolinska University Hospital, Stock- Fathke C, Wilson L, Hutter J, Kapoor V, Smith A, Hocking A, Isik holm, Sweden, 5Biochemistry, Cellular & Molecular Biology, 6 F. Contribution of bone marrow-derived cells to skin: collagen Georgetown University, Georgetown, United States, Tissue deposition and wound repair. Stem Cells 2004; 22:812-822. Engineering, Tehran University of Medical Sciences, Tehran, Iran, Disclosure of Interest: None declared. Islamic Republic Of

Introduction: Skin loss can occur due to many reasons AB013 including burns, diabetic wounds, venous ulcers, trauma, Levels and kinetic of the growth factor release and genetic disorders and etc. Although much progress has been antibacterial activity in allogenic platelet gel for treatment made towards the development of skin engineering but these of GVHD ulcers: a study from the Rome Transplant are not completely efficient and require to improvement. It is Network known that many types of cells including fibroblasts and bone C. Chiara1, F. Angelo Salvatore2,*, L. Alessandro2, P. Alessandra3, marrow stem cells improve performance of composite skin T. Maria Cristina4, B. Cecilia5, F. Eleonora5, A. Maddalena5, substitutes. It is thought that secretion of collagen matrix and D. A. Gottardo3, M. Benedetta3, B. Annalisa3, A. William3, epidermal differentiation takes place after mesenchymal A. Gaspare1 on behalf of Rome Transplant Network epithelial communication. Development of the better trans- 1Immuno-haematology Department, University "Tor Vergata", plantable dermal layers that support keratinocyte proliferation 2Immuno-haematology Department, Policlinico Tor Vergata, is very important for therapeutic option. Here we worked on 3Hemato-Oncology Transplant Unit, University "Tor Vergata", an autologous construct as a successful skin substitute by 4Hemato-Oncology Unit, University "Campus Bio Medico", culture of human Adipocyte Derived Mesenchymal stem cells 5Immuno-haematology Department, University "Tor Vergata", (hADMSC) and human keratinocytes on the Hyalorunic acid Rome, Italy Collagen matrix. Materials (or patients) and methods: Bi-layer skin culture Introduction: Patients affected by acute and chronic graft- was generated by using biodegradable Hyalorunic acid versus-host disease (GVHD) involving skin and oral mucosa Collagen matrix. To analyze the effects of the hADMSC on may develop deep and painful wounds sometimes refractory the epithelial regeneration, keratinocytes were seeded onto to conventional therapies. In our previous clinical study, we the hADMSC-populated matrix and cultured at 37 1C, in a 5% demonstrated the efficacy and safety of the platelet (PLT) gel 6 CO2 and 20% O2 humidified atmosphere. First, 10 hADMSC produced using Vivostat System (Vivolution A/S, Alleroed, were seeded onto the matrix and cultured for 3 days in the Denmark) in the management of GVHD ulcers (Picardi A. et al.

[AB012]

S558 Transfusion. 2010 Feb;50(2):501-6). The Vivostat System is an and compared it to MSCs generated by plastic adherence automated system for the on-site preparation and (PA-MSCs). application of autologous or allogenic derived fibrin sealant Materials (or patients) and methods: In this study, we or PLT-rich fibrin (PRF). In our trial, the gel was prepared compared wound healing potential of CD271-MSCs and from healthy donors. Aim of this study was to assess in vitro, in PA-MSCs of passage 2 and 4. CD271-MSCs and PA-MSCs allogenic PRF, levels and kinetic of the growth factors were generated from bone marrow mononuclear cells (GFs) release and the antimicrobial activity against some (BM-MNCs) isolated from bone marrow aspirates of four bacterial strains. healthy donors. For purpose of this study we used an in vitro Materials (or patients) and methods: After the production of model of wound healing (CytoSelectt 24-Well Wound Healing 5 PRF, 1 ml of each product was dispensed into a flask Assay) from the Cell Biolabs company (BIOCAT GmbH, and then added of 7.5 ml platelet poor plasma (PPP). The GFs Heidelberg, Germany). The wells of the kit contain plastic levels from PPP were evaluated at the time of production (T0) inserts which create a ywound field‘‘ with a defined and at 6, 24, 48 and 72 hours. Using commercial kit (R&D gap of 0.9 mm for measuring the migration rate of System Inc., Minneapolis, MN, USA) through Enzyme-Linked cells. Cell suspensions containing 2x105MSCs/ml DMEM with Immunosorbent Assay (ELISA), the levels of Platelet-derived 1% FCS were cultured until they formed a monolayer around Growth Factor (PDGF-BB), Vascular Endothelial Growth Factor the insert (24 h). The next day, the inserts were removed (VEGF), (EGF) and Transforming from the wells, leaving a precise 0.9 mm open ‘‘wound field’’ Growth Factor-b1 (TGF-b1) were detected. This protocol was between the cells. Cells were monitored under phase applied to assess the levels of the same GFs on auto- contrast and the photographs of the wounded area were logous PRF. To evaluate the antimicrobial activity, taken at 0 h, after 6 h, 12 h and 24 h. To analyse which types of 500 ml of the same products, were applied in the center of cells are better in wound healing, first we compared the different bacterial strain colonies previously seeded wound healing capacity of different passages (P2 and P4) of on plates (Escherichia coli, Pseudomonas aeruginosa, each type of MSCs after 6, 12 and 24 hours. After that we Staphylococcus aureus, Enterococcus faecalis, Staphylococcus compared wound healing capacity between the CD271-MSCs epidermidis and Streptococcus agalactiae). The results and PA-MSCs. were interpreted according to the presence or absence of Results: Our data indicate that in the first six hours of microbial colonies at the application site of the product. experiment no difference in the wound healing potential Following the same protocol, the antimicrobial activity between CD271-MSCs of passage 2 and passage 4 were was also tested using allogenic fibrin sealant obtained by observed. But, 12 and 24 hours after starting the assay CD271- Vivostat System. MSCs of passage 4 had significantly higher wound healing Results: At T0, the levels of PDGF-BB, VEGF and TGF-b1 in the potential than CD271-MSCs of passage 2 (Po0.006 and autologous PRF were higher than in allogenic PRF. The levels Po0.006, respectively). When compared two types of MSCs, of PDGF-BB in autologous PRF are higher up to 48 hours the data indicate that CD271-MSCs as well as PA-MSCs of whereupon the difference tended to shrink. After 6 hours, the passage 2 in initial phases of wound healing, in the first levels of VEGF decreased and began to increase after 24 hours. 6 hours, demonstrated an equal healing potential. Interest- The levels of TGF-b1 remained stable at the different time ingly, CD271-MSCs of passage 2 were significantly more points of observation. The basal levels of EGF were similar, effective than PA-MSCs in closing the wound after 12 hours increased after 6 hours and then remained stable up to and 24 hours of initiation the assay (Po0.03 and Po0,02, 72 hours. PRF was able to inhibit the growth of E. coli, P. respectively). Alsow, CD271-MSCs of passage 4 were superior aeruginosa, E. faecalis and S. aureus but not of S. epidermidis to PA-MSCs of the same passage 12 hours after initiation of the and S. agalactiae. By testing fibrin sealant, no growth assay (Po0.03) inhibition against the same bacterial species colonies was Conclusion: During the wound closure, both CD271-MSCs and observed. PA-MSCs of passage 4 were superior to MSCs of passage 2. The Conclusion: These preliminary results show that allogenic and CD271-MSCs of both passages, however, were more effective autologous PRF have the same kinetic of GFs release and, in wound healing than PA-MSCs, which could make these cells despite their low levels, they are probably sufficient to more suitable for treatment of the wounds. promote tissue regeneration as demonstrated by our previous Disclosure of Interest: None declared. clinical study. Allogenic PRF has an antibacterial activity against the most common bacterial pathogens, which is AB015 completely absent in the fibrin sealant. Tissue regeneration Cardiomyocyitic differentiative potential of umbilical cord and the concomitant ability to inhibit bacterial growth could blood: focus on c-kit þ cells play an important role when PRF is used to care GVHD related M. G. Iachininoto1, E. R. Nuzzolo1, S. Capodimonti1, S. Spartano1, wounds. 1 2 1,* Disclosure of Interest: None declared. M. Bianchi , G. Leone , L. Teofili 1Transfusion Medicine, 2Hematology Department, Catholic University, Rome, Italy AB014 Wound healing potential of mesenchymal stromal cells Introduction: The c-kit oncogene is the main regulator of generated from human CD271 þ bone marrow cardiomyogenesis; the intracoronary administration of auto- mononuclear cells logous c-kit þ cardiac stem cells (CSC) isolated and expanded H. Latifi-Pupovci1,2,*, Z. Kuc¸i1, S. Wehner1, T. Klingebiel1, P. Bader1, from myocardium, was proved beneficial in ischemic cardio- S. Kuc¸i1 myopathy (Bolli et al. Lancet. 2011; 378:1847). Since aging and 1Dept. of Hematology/Oncology, University Children’s Hosital, chronic illnesses significantly limit the stem cell potential of Frankfurt am Main, Germany, 2Department of Physiology and autologous CSC, the search of allogeneic alternative source of Immunology, Faculty of Medicine, University of Prishtina, cells able to contribute to myocardial regeneration may be Prishtina, Albania valuable. Materials (or patients) and methods: We isolated by Introduction: Despite the fact that there are several immunomagnetic method (Miltenyi Biotec) the c-kit þ cell in vivo studies which showed that mesenchymal stromal cells population from cord blood (CB) mononuclear cells (MNC). The (MSCs) have a high impact on wound healing, so far, no c-kit þ cells were characterized for their immunophenotype evidence exists on the healing capacity of different and for the RT-PCR expression of 84 genes related to stem cell MSC-subsets. In this in vitro study, we focused on wound identification, growth and maintenance (PAHS-405Z, Qiagen) healing capacity of MSCs generated from positively selected The myocardial differentiation potential of CB c-kit þ cells was CD271 þ bone marrow mononuclear cells (CD271-MSCs) evaluated in fibronectin coated dishes under three culture

S559 conditions a) alfa-Minimum Essential Medium (MEM) þ 10% were measured in the CBU and CB-PRP: TNC count, RBC count, FBS þ 10-8 M (Beltrami et al. Cell 2003; 114: PLT count, Volume. Sterility of the final product was evaluated 763); b) MEM þ 5% FBS þ cyclosporine-A 3 mg/ml (Fujiwara by BacT/ALERT BPA-PBN (Biomerieux). CB-PRP was activated et al. PLoS ONE 2011, 6:e16734); c) MEM þ 10% FBS þ Bone by Batroxobin and calcium gluconate (Plateltex-Act) to induce morphogenetic protein 4 (BMP4) 100 ng/ml þ TGFb 10 ng/ml PG formation. CB-PRP and CB-PG content of growth factors (Lagostena et al. Cardiovasc Res. 2005; 66:482). We cultured was evaluated on a Luminex instrument using a Bio-Plex Pro the whole MNC fraction in the same conditions, as control. T In Assay (Bio-Rad Laboratories) following manufacturer selected experiments, c-kit þ cells were grown over rat instructions cardiomyocyte layers (Lonza). The differentiation was assessed Results: The characteristics (Volume, PLT, RBC, TNC) of CB by evaluating through RT-PCR and flow cytometry the units before (PRE) and after (POST) processing are reported in expression of the cardiomyocyte lineage markers GATA-4, table 1 together with PLT recovery and concentration, TNC GATA-6, alfa-sarcomeric actin (a-SA), b-myosin heavy chain (b- and RBC depletion in the PRP products. MHC) and cardiac troponin T (cTnT). Automated protocols resulted in a PLT concentration of Results: We found that 0.75±0.15% of CB mononuclear cells 4,82±1,46 fold. The CB-PRP final product had almost no RBC (MNCs) were c-kit þ . We obtained an over 80% pure c-kit þ cell content and very low WBC contamination with a mean PLT population. As compared to whole MNCs, c-kit þ population count of 1,12±0.37x10^9/mL. Mean PLT recovery was was highly enriched with cells expressing CD34 (48% versus 31,41±9,74%. 0.2%) and CD133 (30% versus 0,3%). Variable proportion of Dosage of growth factors before (CB-PRP) and after platelet c-kit þ cells co-expressed the CD45 antigen (33±8%), as well activation with batroxobin (CB-PG) revealed the presence of as various endothelial cell markers (CD54: 28.5±2%, CD105: bFGF, IP-10, VEGF, PDGF-bb and HGF supporting the use of 5±1%; CD144: 1±0.3%; CD141: 2±0.2%), while no co- this emo-component as source of factors suitable for tissue expression of KDR was detected. Both class I and class II HLA repair strategies. No microbial contamination due to CB antigens were detectable in more than 80% of c-kit þ cells. At processing was detected. gene expression profile, CB c-kit þ cells contained significantly higher RNA levels of Aldehyde dehydrogenase-A1, c-Myc and BMP1: these genes are acknowledged markers of immaturity in cells of various lineages. The myocardial differentiation ability of c-kit þ cells was investigated in comparison with whole CB MNCs. After 3 weeks, 13 to 60% of adherent cells recovered from MNCs cultured with either cyclosporin A or BMP 4 and TGFb, expressed GATA-4, GATA-6, a-SA, b-MHC and cTnT: all these cells were c-kit-. In contrast, in adherent cells obtained from cultures of c-kit þ cells, no cardiomyocyte markers were detectable. When c-kit þ cells were co-cultured over rat cardiomyocyte layers (Lonza), a very scarce proportion of cells (1-3%) acquired the expression of GATA-4. Conclusion: Overall, these data confirm that CB contains cell progenitors endowed with cardiomyocyte differentiation capacity. In contrast to myocardium-derived CSC, CB cells able to acquire in culture cardiomyocyte markers are not included within the c-kit þ cell fraction. Supported by MIUR Conclusion: Automated processing of CB for PRP production (PRIN 2010S7CET4). is a fast procedure (20 min automated vs 100 min manual Disclosure of Interest: None declared. procedure), operator-independent and results in a ready-to- use, standardized and safe product for clinical applications. AB016 Further protocol improvements are ongoing to obtain higher Closed automated protocol for platelet rich plasma PLT recovery. CB-PRP and CB-PG contained growth factors production from cord blood involved in the tissue healing process. 1 1 1 2 Disclosure of Interest: B. Mazzanti: None declared, A. Gelli: B. Mazzanti , A. Gelli , L. Ballerini , T. Gorseleben , None declared, L. Ballerini: None declared, T. Gorseleben M. Santosuosso1, S. Urbani1, R. Saccardi1,* 1 2 Employee of: Biosafe, M. Santosuosso: None declared, S. AOU Careggi, FLORENCE, Italy, Biosafe, Eysin, Switzerland Urbani: None declared, R. Saccardi: None declared. Introduction: Platelet rich plasma (PRP) is a platelet concentrate generally produced by autologous blood for topic use. Platelets are rich in growth factors which play a relevant role in tissue healing and PRP as well as its activation product (platelet gel, Gene therapy PG) are widely used in regenerative medicine applications. Cord Blood (CB) is an important alternative source of hematopoietic stem cells (HSC) for allogeneic transplantation and today more than donated 600.000 CB units (CBU) are available worldwide in AB017 CB Banks. However only 10% of collected CBU are eligible for The assessment of MYC and particular microRNAs transplantation due to the low stem cells content. An alternative cooperation with signaling pathways in breast cancer use of the CBU unsuitable for banking would optimize the in the trajectory to reveal osteolytic bone metastasis process and further add a value to the donation. The aim of this V. Pourteimoor1, S. Mohammadi-Yeganeh2,*, M. Yazdani3 study is therefore the standardization of an automated closed 1Biology Department, Faculty of science, University of Zanjan, system to produce PRP from CB. P.O. Box 313, Zanjan, I.R., Iran, 2Cellular and Molecular Biology Materials (or patients) and methods: The efficacy of Research Center, Biotechnology Department, Shahid Beheshti automated protocol for the processing of whole cord blood University of Medical Sciences, Tehran, Iran, 3Biology Depart- units to produce PRP, using Sepax 2 device (Biosafe, Switzer- ment, Faculty of science, University of Zanjan, P.O. Box 313, land) was evaluated. Zanjan, Iran, Islamic Republic Of 13 CB units with TNCo1.5 x10^9 and PLT content4150x10^9 were processed with dedicated protocols (PRP v201 and Introduction: During distinct stages of cell development and PRPv203) within 48 hours after collection. Both protocols differentiation, several cellular processes perfectly coordinate consisted of one centrifugation. The following parameters together. MYC as a transcription factor has a fundamental role

S560 in this landscape. In addition to, diverse signaling pathways support growth of Mesenchymal Stem Cells (MSCs) in construct the substantial interaction networks with MYC and serum free or serum low conditions. We have previously miscellaneous arrays of miRNAs in the connection to manifest found an inverse correlation between the expression of PDGF- heterogeneous patterns of breast cancer progression and R on Mesenchymal Stem Cells (MSCS) and PDGF-BB in BM bone marrow metastasis. The large number of signaling plasma of the endosteal and central regions of the bone molecules implicates in aberrant expression of MYC, which in marrow (BM) niche. Addition of PDGF-BB to culture medium turn, can impact on the incidence of invasive tumor containing 10% serum resulted in a dose dependent inhibition phenotype through regulatory processes such as proliferation, of proliferation of MSCs and downregulation of PDGF-Rb survival, and invasion. This review concentrates on MYC expression by MSCs. Here, we used Mesylate (IM), oncogene, specific miRNAs and unreconstructed/critical sig- which inhibits the tyrosine kinase domains of PDGF-R, c-kit naling pathways in the maintenance of malignant character- and abl, as well as two specific PDGF-R inhibitors, IV (PRI-IV) istics of breast cancer cells at the expense of haematopoietic and V (PRI-V) to assess if PDGF-BB modulates its effect on MSC stem cells (HSCs). Various levels of signaling mediators proliferation through differences in cell cycle activity and contribute to colonization of breast cancer cells in secondary apoptosis. tumor mass through normal cell exhaustion. The examples of Materials (or patients) and methods: BM from healthy these mediators are prolyl-isomerase pin1, P53, NF-kB, TNFa, donors was collected for transplantation purposes. Remaining hedgehog, ErbB signaling compartments, Notch and Wnt cells were used for culture of MSCs up to passage 3 in DMEM/ pathway effectors, TGF-b, Brca1, Src and Jag1, 2. MCDB-201 supplemented with 10% fetal calf serum (FCS). Materials (or patients) and methods: We studied and Effects of PDGF-BB, IM and PRI-IV/PRI-V on proliferation of evaluated disparate articles on various analytical foundations MSCs under serum supplemented (10% FCS) and low serum of the desired gene and compared the methods of these conditions (2% FCS) were assessed real time using xCELLi- disquisitions in order to investigate the role of MYC oncogene, gence (Roche) technology, based on the measurement of MSC miRNAs and the signaling pathways for giving the overall impedance. Effects of PDGF-BB, IM and PRI-IV/V on MSC cell result of MYC activity on metastasis. cycle were assessed using Propidium Iodide (PI) and measured Results: Our comparisons demonstrated that specific recep- using a FACSARIA (Becton Dickinson). Apoptosis was assessed tors, specially frizzled receptors, HR, toll like receptors, and using Annexin-V/PI. E-cadherin whose interfere with peculiar signaling pathways Results: MSCs displayed decreased proliferation under low that can influence the expression levels of MYC oncogene and serum conditions (2% FCS) in comparison to normal serum miRNA-200, Let7, miRNA-301a, miR-33b, miRNA-191 that have levels (10% FCS). Increasing doses of PDGF-BB, resulted under crucial roles in the initiation, maintenance, progression and both culture conditions, in suppression of proliferation with metastasis of breast cancer cells and their biological viability in doses 45 ng/mL resulting in irreversible suppression of different situations. proliferation. Similar inhibitory effects on proliferation of MSCS Conclusion: The feasibility of using these interactions for were obtained when IM, PRI-IV and PRI-V were added in introducing an effective treatment appertains to multiple factors. increasing doses. Combined use of PDGF-BB/IM/PRI-IV/PRI-V In the vivo conditions, systemic effects of several factors such as resulted in an even more profound suppression of MSC oncogenes, hormones, signaling mediators, and also epigenetic proliferation. Cell cycle analysis after 24 hours treatment with effectors for example stress or methylation levels involve in the PDGF-BB, IM and PRI-IV/V demonstrated arrest of MSCs, formation of specific and sophisticated cellular processes. treated with IM and PRI-V in G0/G1 phase, no effect of PRI-IV Nevertheless, independent or intervention scrutiny of the on cell cycle activity and a stimulating effect of PDGF-BB on authentic breast cancer models with the devastating metastasis cell cycle activity. PDGF-BB, IM and PRI-V did not affect to the bone marrow that its cause can be dysregulation of MYC apoptosis, whereas PRI-IV increased levels of Ann-V positive activity is important to develop new therapeutic strategies. cells twofold. References: 1. Brabletz, Simone, et al. "The ZEB1/miR-200 Although PDGF-BB is a known mitogen for MSCs under serum- feedback loop controls Notch signalling in cancer cells." The free conditions, we found a dose-dependent inhibition of EMBO journal 30.4 (2011): 770-782. proliferation by PDGF-BB and different PDGF-R inhibitors 2. Yun, Jieun, et al. "Signalling pathway for RKIP and Let-7 under serum-supplemented conditions. Our results suggest regulates and predicts metastatic breast cancer." The EMBO that overdoses of PDGF-BB downregulate PDGF-R expression journal 30.21 (2011): 4500-4514. on MSCs, and slow down proliferation; whereas IM and PRI-V 3. Zhang, Xiang H-F., et al. "Latent bone metastasis in breast appear to reduce PDGF-R signalling, prevent cells from cycling cancer tied to Src-dependent survival signals." Cancer cell 16.1 and proliferation and PRI-IV decreases MSC numbers through (2009): 67-78. induction of apoptosis. 4. Xing, Fei, et al. "Hypoxia-induced Jagged2 promotes breast Conclusion: In presence of serum, PDGF-BB inhibits cancer metastasis and self-renewal of cancer stem-like cells." proliferation of MSCs through downregulation of its Oncogene 30.39 (2011): 4075-4086. receptor. Imatinib and PDGF-R inhibitor V induce cell cycle Disclosure of Interest: None declared. arrest of MSCs, whereas PDGF-R inhibitor IV induces apoptosis of MSCs. Disclosure of Interest: None declared.

Non-hematopoietic stem cells AB019 Out of the bag – recovering fully functional cells from usually discarded bags S. M. Lopes1,*, F. Amado1, S. Ferreira1, A. F. Bordalo1, S. Roncon1 AB018 1Cellular Therapy, IPO PORTO, Porto, Portugal Effects of Platelet Derived Growth Factor (PDGF) and PDGF-R inhibitors on proliferation, apoptosis and cell cycle Introduction: Bone marrow (BM) is the primary source of of Mesenchymal Stem Cells progenitor cells in the adult body. First recognised as home to G. Guney1,2, F. Aerts-Kaya1,2,*, D. Uckan-Cetinkaya1,2,3 haematopoietic progenitor cells (HPC), it also contains progenitor 1Center for Stem Cell Research and Development, 2Institute of cells of other lineages, such as mesenchymal and neurological. Health Sciences, Hacettepe University, 3Department of Pediatric Although its use in the Haematopoietic Stem Cell Transplant Hematology, Hacettepe University Medical Faculty, Ankara, Turkey (HSCT) setting has decrease in last decades, its interest in cellular therapy is increasing, with numerous protocols using BM derived Introduction: Platelet Derived Growth Factor-BB (PDGF-BB) is mononuclear cells (MNC), such as ex vivo expansion of HPC, a potent mitogen for cells of mesenchymal origin and used to neuro and mesenchymal stem cell differentiation.

S561 The standard protocol for BM collection is thru multiple irradiation, low platelet count before mobilization and low aspirations in the iliac crests, added in a collection bag CD34 þ cell count in peripheral blood(PB)(o20/mL) on leuko- containing an anticoagulation solution (ACD-A). Prior to cyte recovery. Different guidelines were published to optimize infusion or manipulation, the collected BM must be filtered mobilization procedures particularly with new approaches to remove bone fragments and cell clumps. including for a CXCR4 inhibitor that improved In order to obtain a minimum MNC necessary for peripheral blood stem cell (PBSC) mobilization when com- most protocols, a relatively large volume (420 ml) of BM is bined with granulocyte colony stimulating factor(G-CSF). A collected or removed from BM graft. To overcome this position statement from the EBMT proposed the administra- difficulty, we studied whether cells that remain in the tion of plerixafor preemptively in pts with CD34 þ cell count collection bag and filter system, should not be considered o10/ mL on leukocyte recovery.Nevertheless, no clear position clinical waste. statement was reported for pts with CD34 þ cell count in PB Materials (or patients) and methods: From 14 BM grafts between 10 and 20/ mL, considered in a ‘‘grey zone ‘‘.( Bone collected at our institution, intended for allogeneic Marrow Transplant 2014; 1-8).The objective of our study was to HSCT, we compare the amount of MNC obtained from 1 ml assess the feasibility and efficacy of plerixafor to induce of the filtered BM (used for clonogenic assays) and from the efficient collection of PBSC in pts with MM and relapsed L who waste system, and calculated the virtual equivalence in BM were in the ‘‘grey zone’’ after mobilization with chemotherapy volume. (CT) þ G-CSF. The collection bag and filter system were back-washed Materials (or patients) and methods: Eight pts were and rinsed with RPMI under aseptic conditions. MNC included.The decision of adding plerixafor was based on for both samples were isolated by density gradient CD34 þ cell count which was between10 and 20/ mL centrifugation. on leukocyte recovery.One dose of plerixafor (0.24 mg/Kg) The viability of the recovered MNC was evaluated by trypan subcutaneously (S/C) was planed followed by one apheresis blue exclusion and functionality evaluated by in vitro differ- session with a target to collect more than 2.106 /Kg of entiation into mesenchymal stem cells (MSC). CD34 þ cells. MSC were obtained from long time culture of MNC in DMEM Results: three of the 8 pts were treated for MM and media supplemented with foetal bovine serum, at 371C and 5 for L. Median age was 35 yo (19-54). There were 5 females 5% CO2. and 3 males. All the pts received more than 2 lines of Results: From the 14 BM products we obtained an average of treatment which included fludarabine in one patient (pt), 1.18x108 recovered MNC. After adjusting for the MNC isolated melphalan in 2 pts and lenalidomide in 3 pts. All the pts from the 1 ml filtered sample, we can estimate this recovery is had a bone marrow cellularity d30% and 6 had thrombo- the equivalent to starting with an average 51 ml of filtered BM. cytopenia at mobilization. CT ( þ G- CSF) used for In all cases viability was superior to 90%, and long term MSC mobilization was cyclophosphamide in the 3 pts with MM, cultures were established, as shown by morphology and DHAP (dexamelhadno, cytarabine, cisplatin) in 4 pts and immunophenotype. IGEV (ifosfamide, gemcitabine, vironelbine) in 1 pt.The median Conclusion: With this study we demonstrated that the usually no of CD34 þ cells in PB before plerixafor injection was wasted BM collection bag and filter system is a source of MNC 14/ mL(11-15).After plerixafor injection an increase in CD34 þ to consider in the investigation and clinical settings, capable of cell count in PB was seen in 6 pts, while CD34 þ cell count yielding large amounts of viable, fully functional and sterile remained between 10 and 20/ mL in 2 pts. The median time MNC. from CT to plerixafor injection was 16 days(14-24).All the pts Disclosure of Interest: None declared. reached the target no of CD34 þ cell collected by one apheresis with a median of 3.4 106 /Kg (2.8-6.8). Plerixafor was well tolerated Conclusion: Our results show that one dose of plerixafor given ‘‘ on demand’’ for the pts with MM and relapsed L who were in Stem cell mobilisation and graft the ‘‘grey zone’’ after mobilization with CT þ G-CSF can induce engineering efficient collection of PBSC independently of the no of CD34 þ cells in PB obtained after prelixafor. Disclosure of Interest: None declared. AB020 Efficient collection of peripheral blood stem cells with one AB021 dose of plerixafor in patients with myeloma and Comparison of the Efficiency of High and Low Dose lymphoma with CD34 þ cell count in peripheral blood Cyclophosphamide for Stem Cell Mobilization between 10 and 20/lL on Leukocyte recovery after A. Birekul1,*,A.U¨nal1,M.Ç.U¨nal1, E. E. Turak1, G. Akyol1, mobilization with chemotherapy and GCSF N. Mandacı1, E. Yıldızhan1, E. Karakus¸1, M. Solmaz1,M.O¨ ztekin1, A. O. Ibrahim1,*, A. Mugharbil2, A. Youssef3, R. Jalloul3, G. Nsouli3, S. S¸ıvgın1, L. Kaynar1, B. Eser1, M. Çetin1 T. Jisr4 1Hematology, Erciyes University, kayseri, Turkey 1Hematology/Oncology, Makased University Hospital, 2Hematology /Oncology, 3Hematology/Oncology, 4Laboratory Introduction: In this study, we compared the effectiveness Medicine and Blood Bank, Makassed University hospital, Beirut, and side effects of high dose and low dose cyclophosphamide Lebanon (2,4 gr/m2 and 4 gr/m2) on stem cell mobilization in patients with multiple myeloma and lymphoma whose planned Introduction: High-dose chemotherapy (HDCT) followed by autologous stem cell transplantation. autologous peripheral stem cell transplantation (APSCT)is an Materials (or patients) and methods: Stem cells are mobilized effective therapeutic strategy for patients (pts) with multiple with high dose (4 gr/m2) or low dose (2,4 gr/m2)cyclopho- myeloma (MM) and relapsed lymphoma(L). A successful APSCT sphamide and G-CSF (Filgastrim and Lenogastrim) in patients is dependent on a sufficient amount of CD34 þ cells transfused with multiple myeloma and lymphoma, between 2011-2013 in for a durable engraftment. The minimal required number (no) Apheresis Unit of Erciyes University, Stem Cell Transplantation of CD34 þ cells for APSCT is 2.106 /Kg. Although the majority Center. We evaluate the efficiency of cyclophosphamide doses of pts are able to mobilize sufficient CD34 þ cells, approxi- on stem cell mobilization and collection in patients. mately 15% fail to do so. Several factors were reported as Results: In this study 56 patients which 41 male and 15 female predictive of poor or failed mobilization such as old age, were enrolled. 29 patients were multiple myeloma, and 27 of advanced stage disease, high no of prior treatment lines, type them were lymphoma. Median age was 55 (25-71). Cyclopho- of prior therapy (fludarabine, melphalan, lenalidomide.), prior sphamide 4 gr/m2 were applied to 30 patients while 2,4 gr/m2

S562 were applied other 28 patients for stem cell mobilization. application was subcutaneously. The median number of Before the process, the amount of median WBC and Plerixafor injections was 1(1-3). CD34 þ cells count were 59,9 103 m/L and 35,5 m/L, respec- Results: In all patients, the peripheral blood CD34 þ cell tively. After apheresis, median amount of CD34 þ cells were counts increased. The median CD34 þ cells count after first 6,89 106/kg in the patients given low dose and plerixafor dose was 36 mL (20-80) with the median increase of 9,88 106/kg in the patients given high doses (P40,05). 7-fold (3 to13). The total number of WBCs also increased but It was found that median amount of mononuclear cells were less markedly (2-4 times). Five patients underwent 1 apheresis, 1,13 108/kg and 1,04 108/kg in low and high doses, 10 patients 2, and 3 patient 3 aphereses. Median collected respectively (P40,05). Cyclophosphamide 4 gr/m2 were CD34 þ cells from one apheresis was 3,25x106/kg (1, 3–12, applied to 15 of 29 patients that diagnosed multiple myeloma. 6 106/kg). Fifteen patients were transplanted successfully Median amount of CD34 cells were 9,32 106/kg after a period of different length. None of the patients had and mononuclear cells were 1,05 106/kg in low dose serious complications after Plerixafor. cyclophosphamide applied patients. It was detected that Conclusion: Based on this report and the reviewed median amount of CD34 cells were 11,18 106/kg and literature, we conclude that the ‘‘pre-emptive’’ application of mononuclear cells were 1,01 108/kg in high dose given Plerixafor is efficiency and save procedure. It also patients. There was no statistical significance between CD34 þ offers considerable financial benefit in comparison with cells and mononuclear cell count of low dose and high dose ‘‘standard’’ design due to a minimal number of mobilization applied multiple myeloma patients. Cyclophosphamide 4 gr/ and PBSC collection and in time conducted myeloablative m2 were applied to 13 of them while 2,4 gr/m2 were applied therapy. other 14 lymphoma patients. CD34 þ cell count was Disclosure of Interest: None declared. 5,10 106/kg in low dose and 8,85 106/kg in high dose applied patients while the number of mononuclear cells were 8 8 AB023 1,69 10 /kg and 1,17 10 /kg in low dose and high dose Peripheral blood stem cell mobilization with pegfilgrastim applied patients, respectively. in lymphoma patients Conclusion: We did not detect any significant difference N. Gredelj Simec1,*, V. Zatezalo1, D. Radic Kristo1, S. Jurenec2, between the number of collected CD34 þ cells in low and high 3 1 1 dose cyclophosphamide given lymphoma and multiple mye- M. Lukic , N. Zovko , S. Ostojic Kolonic 1Department of hematology, 2Transfusion medicine, Clinical loma patients. However, we suggest stem cell mobilization with 3 low dose cyclophosphamide is safer and sufficient, considering Hospital ’’Merkur‘‘, Clinical transfusion, Croatian Institute of that high dose cyclophosphamide cause longer and more transfusion medicine, Zagreb, Croatia aggressive neutropenia resulting in infections, frequently. Disclosure of Interest: None declared. Introduction: is a recombinant human granulocyte colony stimulating factor (GCSF) and pegfilgrastim is its pegylated form (pegGCSF). Both are used in prevention of AB022 febrile neutropenia and can be used in stem cell mobilization. Efficacy and safety of pre – emptive mobilization The aim of our study is to compare the efficacy of a single of peripheral blood progenitor cells for autologous 6 mg dose of pegfilgrastim with daily administration of transplantation with Plerixafor filgrastim for stem cell mobilization and to compare time to B. E. Avramova1,*, M. Yordanova1, T. Hristova1, D. Konstantinov2 engraftment after autologous stem cell transplant in lym- 1Transplantation, 2SPECIALIZED HOSPITAL FOR PEDIATRIC phoma patients. ONCOHEMATOLOGY, Sofia, Bulgaria Materials (or patients) and methods: From January 2014 to November 2014 we harvested peripheral blood stem cells Introduction: Myeloablative chemotherapy with autologous (PBSC) of 40 patients with different lymphoma subtypes peripheral blood stem cell (PBSC) support is a salvage (group 1), while 15% (N ¼ 7) patients failed to mobilize. In this treatment option for many patients (children and adults) with group we used pegfilgrastim for stem cell mobilization after hematological malignancies and solid tumors. A proportion of chemotherapy. Group consisted of 25 male and 15 female these patients have ‘‘failure ‘‘after mobilization. Plerixafor patients with median age of 52,5 years (range 20-70). Most augment PBSC collection in these patients, but the optimal patients (32,5%) had Hodgkin’s disease, followed by DLBCL approach for incorporating it into mobilization is uncertain. It (25%), mantle cell lymphoma (15%), T-NHL (15%) and follicular has been studied mainly in ‘‘classical’’ application with G-CSF lymphoma (12,5%). Mobilization regimens were mainly mini- after first unsuccessful mobilization. Pre-emptive or ‘‘on BEAM and ICE (70% and 20%, respectively), while 10% of demand’’ use of Plerixafor is efficient and cheaper, but the patients recieved other regimens. In this group 70% of time of administration and patient selection are under patients underwent autologous PBSCT until November 2014 investigation. and all recieved pegfilgrastim 6 mg subcutaneous on day 1 We evaluated the efficacy, safety and financial benefit of ‘‘pre- after transplantation. We compared this group to a historical emptive’’ use of Plerixafor after chemomobilization in patients cohort (group 2) of 126 lymphoma patients (65 male, 61 who seem to mobilize poorly. female), median age 45 (range 19-71) who had similar Materials (or patients) and methods: Between March 2010 distribution of lymphoma subtypes. In this group 90% of and October 2014 eighteen patients (9 adults and 9 children) patients recieved miniBEAM as priming regimen, the rest with different types of cancers were mobilized with Plerixafor recieved other regimens. All patients in this group were in our hospital, due to ‘‘mobilization failure’’ (CD34 þ cells transplanted and recieved filgrastim 5 mcg/kg daily starting on o10/mL at leukocyte recovery after chemotherapy and G-CSF, day 1 until resolution of neutropenia. BEAM regimen was used or poor yield from the first collection). In 11 patients (7 adults as conditioning regimen before autologous PBSCT in both and 4 children) Plerixafor was used ‘‘pre-emptive’’ (before the groups. second leukapheresis, after mobilization with chemotherapy Results: Patients in a group 1, using miniBEAM as priming and G-CSF, due to very low collected cells count after the first). regimen, had CD34 þ cells mobilized on median of 17,5 days The median patient’s age was 22, 9 years (4-44).Ten patients (range 13-24, average 17,3). Using ICE as priming regimen, were with Hodgkin disease, 2 with NHL, 2 with neuroblastoma, median time to mobilize was 14 days (range 11-16, average 2 with germ cell tumors and 2 with Ewing sarcoma. Six 13,5). In group 2, using miniBEAM as priming, median time to patients have had previously one and 1 patient two mobilization was 19 days (range 15-34, average 19,4), while unsuccessful mobilizations and for 11 patients (‘‘pre-emptive using ICE median was 16 days (range 14-22, average 17,3). ‘‘group) this was the first mobilization. The median CD34 þ There is no statistically significant difference between two cells count in the blood before Plerixafor application was 6/ mL groups in time to stem cell mobilization using the above (1-18). The single dose of plerixafor is 240 mg/kg and the stated regimens. Median time to engraftment of leukocytes

S563 and thrombocytes was 10 days in group 1, while in group 2 Stem cell source median time to engraftment of leukocytes was 10 days and for thrombocytes it was 11 days. There is no statistically significant difference. Conclusion: Our study has shown no significant AB025 difference between single fixed dose of pegfilgrastim versus Significant increase number of WJ-derived mesenchymal multiple daily administrations of filgrastim in time to stem cells resulting in international cooperation in regenerative medicine mobilization, as well as in time to leukocyte and thrombocyte 1,* 2 3 engraftment after autologous PBSCT. While pegfilgrastim does B. Roxana , P. Dorin on behalf of 1, B. Dariusz on behalf of 2, O. Tomasz3 on behalf of 2, M. Magdalena3 on behalf of 2, not seem to be superior in stem cell mobilization and in 3 3 acceleration of engraftment after PBSCT, it is more convenient, C. Iwona on behalf of 2, O. Artur on behalf of 2 1company, S.C. BIOGENIS S.R.L., 2company, S.C. BIOGENIS S.R.L, as it does not require multiple daily administrations as 3 filgrastim does. Bucharest, Romania, company, The Polish Stem Cell Bank, Disclosure of Interest: None declared. (PBKM) S.A., Warsaw, Poland Introduction: The scientists are anticipating that stem cells will revolutionize medicine in the coming years. As we extend the studies on the structure and functions of these cells, thus being Apheresis stem cell collection and able to understand them, stem cells will play an important role processing in treating many diseases, by repairing or replacing damaged tissues with newly developed, healthy ones. We have published numerous working protocols on isolating mesenchymal stem cells from the umbilical cord and on AB024 evidence of morphology and their action as stem cells. Can we use an ultrasound device to improve peripheral In this study we describe first cooperation between European punctures for apheresis stem cell collection procedures? laboratories, which resulted in clinical application for the first B. Wauben1, R. Senden 1,* intrathecale infusion into a Romanian pediatric patient. 1University Hospital Maastricht, Maastricht, Netherlands Materials (or patients) and methods: After parents’ informed consent, we (at Biogenis, the Romanian Laboratory) obtained Introduction: Can we use an ultrasound device to improve and compared 5 samples of umbilical cord, using specially peripheral punctures for apheresis stem cell collection designed separate set for this type of tissue during natural procedures? deliveries and caesarean sections. R. Senden (nurse) en B. Wauben (quality manager), University After visual inspection of each umbilical cord, Wharton’s Jelly Hospital Maastricht, 04-11-2014 tissue was sectioned in order to start the culture. Cells were Introduction Venous access for an apheresis stem cell grown in an incubator at 37 1C, 5% of CO2 and 95% of collection procedure is obtained by placing a cannula humidity in a xeno-free medium, designed for human inside a vein (cannulation). If the vein is not visible and not mesenchymal stem cells. felt (for example by edema, deep veins and obesity) vascular Mesenchymal stem cells were collected from the second access is obtained through central venous access including the passage. In 5 mL of sample volume, there were 30 million cells, risks of complications of a central venous catheter such as which were used for the first intrathecal infusion in a hematomas, pneumothorax, hemothorax and bacterial Romanian pediatric patient. infections. After collection, cells were frozen using cryo-protective Aim To explore whether successful peripheral vascular substance. The cell concentrate solution containing mesench- access is enhanced by the use of an ultrasound device. ymal stem cells was frozen in a controlled manner to a certain temperature level and then placed in the liquid nitrogen Can the echo-images support the nurse, in addition to o visualization and palpation, in achieving a successful vapors at a temperature level below -150 C. cannulation? In addition, viral tests were performed on mother’s peripheral Materials (or patients) and methods: Method blood to determine the probable presence of viruses and the - Instruction staff how to use an ultrasound device final decision was made as to keep the umbilical cords from - The use of an ultrasound device in patients/donors with the negative samples. visible and palpable veins Also, tests were performed to confirm the absence of - Once the nurse knows how to use the new technique, microorganisms, such as Mycoplasma and endotoxins, situ- can be started with obtaining peripheral vascular access by ated in Wharton’s Jelly. patients/donors with veins that are not visible and not We also tested, using flow cytometry, the immunophenotype palpable (because of for example edema, deep veins or of cells. The cells were CD105, CD90, CD73 positive and obesity) negative for CD34, CD45, CD14, CD19, HLA-DR surface Results: Results antigens. Using an ultrasound device has the following advantages: Transportation of the cells from Romania to Poland was 1. If the vein is not visible and not felt, using an ultrasound performed in a state of deep freeze, after having the final device still leads to peripheral vascular access for an apheresis approval from The Romanian National Transplantation Agency. stem cell collection procedure in all cases The transport was performed in special containers with 2. The nurse themself realizes the peripheral vascular access continuous temperature monitoring. Conclusion: Conclusion The new method increases the Results: The presence of mesenchymal stem cells was patient safety. An invasive surgery on the donor (placing a confirmed in PBKM’s laboratory in Poland. central venous catheter) is not necessary. If the vein is not More cultures were established and there was a significant visible and not palpable, ultrasound images still lead to increased number of MSCs in the every next passage. peripheral vascular access. Additional mesenchymal stem cells collection procedure was Discussion performed resulting in new number of mesenchymal stem - The cost of buying an ultrasound device cells after the fifth passage. - Peripheral vascular access takes longer because of the use of This cooperation has completed with the infusion into the first an ultrasound device Romanian pediatric patient with neurologic disease. Disclosure of Interest: None declared. Conclusion: This report indicates that Wharton’s Jelly is a good PRIMARY source for mesenchymal stem cells, which can be isolated, cryopreserved and used in clinical applications.

S564 Results presented in our studies suggest that collected WJ- AB028 MSC may offer a high number of cells, allowing to increase the A retrospective study of the chemomobilization effect potential of mesenchymal stem cells in regenerative medicine. of AAE regime plus rhG-CSF in acute leukemia patients Disclosure of Interest: None declared. J. shi1,*, Y. Luo2, Y. Tan2, H. Huang2 1The first affiliated hospital, 2Bone Marrow Transplantation Center, the First Affiliated Hospital, Zhejiang University School of AB026 Medicine, Hangzhou, China The effective parameters on peripheral blood stem cell apheresis in children Introduction: Explore stem cells mobilization and disease E. Atas1,*, V. Kesik1, O. Babacan2, N. Korkmazer3 consolidation effect of chemotherapy regime:cytarabine, 1Department of Pediatric Oncology, 2Department of Pediatric aclacinomycin and etoposide (AAE) plus recombinant human Hematology, 3Department of Pediatrics, Gulhane Military Medical granulocyte-colony stimulating factor (rhG-CSF) in acute Academy, Ankara, Turkey leukemia (AL) patients before transplantation. Materials (or patients) and methods: During January 2010 to Introduction: Peripheral blood stem cell collection (PBSC) august 2014, 31 AL patients were admitted to our transplanta- sometimes may not be succesfull to collect adequate tion center to keep autologous stem cells. Among these CD34( þ ) stem cell. Thus, we aimed to evaluate the effective patients, 13 patients were acute lymphocyte leukemia(ALL) parameters on amount of CD34 ( þ ) stem cell in children and and other 18 patients were acute myeloid leukemia(AML).All difficulties of apheresis. patients were acquired complete remission and received AAE Materials (or patients) and methods: Medical records of 54 chemotherapy regimen plus rhG-CSF(300ug/day). Stem cell children were reviewed who underwent PBSC apheresis. The apheresis was scheduled when white blood cell(WBC) count changes in CD34 per kg 106 according to day of apheresis recovered greater than 2.0x10E9/L and platelet was more than (Group 1: day 3, Group 2: day 4) with the effects of some 20x10E9/L in peripheral blood. The mobilization and con- factors as disease type, gender and age were investigated. solidation efficiency was analyzed retrospectively. Results: Characteristics of the PBSC as CD34 per kg 106 were Results: 31 patients after chemomobilization regime,the þ 2.1 106/kg in Group 1 and 2.8 106/kg in Group 2, median number of CD34 cells was 2.728 10*6/kg, the respectively. Change in CD34 per kgx106 by time between ALL patients achieved significantly high median number of þ Group 1 and Group 2 was found as significant (P ¼ 0.046). No CD34 cells(5.80610*6/kg) than AML patients(2.278 10*6/ 6 kg,P ¼ 0.037). 19 patients(61.29%) acquired satisfactory mobi- interaction was found between CD34 per kgx10 and disease þ type, gender and age, respectively (P 0.903, P 0.942, lization efficiency (CD34 cells Z2.0x10E6/kg) and 12 ¼ ¼ þ P ¼ 0.999). The mean amounts of CD34 per kg 106 were patients(38.71%) had poor mobilization(CD34 cells 3.01 in o10 years old and 2.24 in Z10 years old at day 3 and o2.0x10E6/kg). During the poor mobilization patients, the 3.3 in o10 years old and 2.7 in Z10 years old at day 4. The percent in AML patients (8patients,44.4%) was significantly mean amounts of CD34 per kg 106 were 3.05 in males and high than ALL patients(4 patients,30.7%). However, there were 1.92 in girls at day 3 hours and 3.45 in males and 1.94 in girls at 2 ALL patients relapsed after AAE chemotherapy. The IV grade day 4. The mean amounts of CD34 per kg 106ofEwing, bone marrow suppression after chemotherapy was neuroblastoma and Hodgkin’s lymphoma were 2.48, 2.88, 2.53 7.15±2.185 days and the median days between chemother- at day 3 and 2.81, 3.31, 2.52 at day 4. We did not find apy end to stem cells apheresis was 15 day. During the bone differences on age (P ¼ 0.296), gender (P ¼ 0.075) and CD34 morrow suppression, 70.97% patients had febrile neutropenia per kgx106 (P ¼ 0.768) levels according to disease types. and need platelet transfusion. Conclusion: Although technically similar to adult apheresis Conclusion: AAE plus rhG-CSF is an effective chemomobiliza- tion regime in AL patients and in ALL patients can achieve procedures, some physicians that have little experience in þ pediatric apheresis worry about PBSC harvesting in children. more CD34 cells. But in ALL should be more careful to Catheter problems such as bleeding, occlusion, thrombosis, choose AAE because the ALL had high risk to relapse. vomiting, nause, skeletal muscle contraction and laughing Disclosure of Interest: None declared. owing to citrate related hypocalcemia, hypomagnesemia, transient trombocytopenia, anemia, hypotension were the AB029 complications of procedures. The amount of CD34 per kg in Changes in the lipid levels during the mobilization PBSC harvesting at day 4 was better than day 3. Disease types, processes age and gender were not important on the CD34 results. M. Comert Ozkan1,*, I. Yavasoglu2, I. Yildirim Simsir3, Disease types were not found significant on the CD34 results. B. Sarer Yurekli3, G. Basol4, C. Kabaroglu4, F. Saygili3, But toxicity and duration of chemotherapy regimen can affect Z. Bolaman2, M. Tombuloglu1, A. Donmez1 the results. Age was not found as effective indicator on CD34 1Hematology, Ege University,˙ Izmir, 2Hematology, Adnan Men- count, but bone marrow capacity is better in younger ages. In deres University, Aydın, 3Endocrinology, 4Clinical Biochemistry, our study the mean level of CD34 was higher in younger ages Ege University,˙ Izmir, Turkey than elder ages. This may explain the lower levels of CD34 levels in subjects with Hodgkin lymphoma who were above 10 Introduction: The results of retrospective human studies that years old with the rate of 80% in this study. However, some evalute the contribution of cholesterol levels in stem cell additional factors may be effective on apheresis. If there is a mobilization obtained different results. Determining the lipid history about multiple and toxic chemotherapy regimen in level changes in the process of mobilization of healthy donors primary or relapsed protocol, we will face to face more and revealing the possible interaction with the mobilization difficulties to perform PBSCA. Factors like priming, risk of process was aimed. hypocalcemia, poor venous access, thrombotic side effects Materials (or patients) and methods: The preliminary have to be evaluated before PBSCA. PBSCA can become a results of our prospective another study that is supported by routine medical procedure in pediatric oncology. Collection of Turkish Hematology Association (Project number: 2013/5) PBSC apheresis appears to be less traumatic than bone was provided. Total cholesterol, LDL cholesterol, HDL marrow harvest. cholesterol and TG levels and periferal blood CD34 þ cell Disclosure of Interest: None declared. levels (BD facsari cell sorter BD Biosiences) in three periods in mobilizing donors (before filgrastim [period I] before apheresis [Period II] and apheresis after the first week [Period III]) were AB027 determined. Abstract Withdrawn Results: Ten (mean age:38.7, M/F:9/1) donors were mobilized with filgrastim (10 mg/kg/day). Peripheral blood CD34 þ cell

S565 level was 1.13±0.53/mL in period I, 34.9±8.52/mL in period II, pseudoaneurysm of the gluteal artery in the setting of and 0.6±0.15/mL in period III. There was no significant enlarging hematoma in a patient following BM harvesting, difference in the levels of total cholesterol (212.6±15.59, because if recognized early, interventional radiology-guided 194.±17.02 ve 218.2±14.08 mg/dL, respectively, P40.05), embolization can lead to immediate and effective resolution of LDL cholesterol (125.8±12.33, 112.3.±12.92 and the problem. This case serves as reminder that even generally 126.8±0.87 mg/dL, respectively, P40.05), HDL cholesterol considered safe, bone marrow harvesting is a procedure with (41.8±1.88, 35.6±3.36 and 42.2±2.02 mg/dL, respectively, potentially severe complications. P40.05), and triglyceride (245.8±33.19, 232.5±34.21 and Disclosure of Interest: None declared. 254.2±43.9 mg/dL, respectively, P40.05) during and after the mobilization process. AB031 Conclusion: According to the preliminary results of our CD34 þ count and not total nucleated cell count has ongoing study, there was no significant changes that may impact on OS in adult dUCBT indicate the possible effects of lipids on the mobilization with N. Collins1,*, R. Whittle1, K. Robertson1, R. Lunn1, K. Sills1, A. Bloor2, filgrastim. 3 1 Disclosure of Interest: None declared. J. Snowden , T. Key 1Histocompatibility and Immunogenetics, NHSBT, Sheffield, 2Stem Cell Transplantation, The Christie NHS Foundation Trust, AB030 Manchester, 3Haematology, Sheffield Teaching Hospitals NHS Unexpected complication of bone marrow harvesting: Trust, Sheffield, United Kingdom considering pseudoaneurysm of the superior gluteal artery Introduction: Umbilical-cord blood transplantation (UCBT) is M. M. Greco1,*, A. M. Carella1, E. Merla1, G. Ciccarese2, an alternative treatment modality for adult patients lacking a N. Cascavilla1 suitable HLA matched sibling or unrelated donor. A sufficient 1Hematopoietic Stem Cells Transplantation Unit, 2Interventional haematopoietic progenitor cell dose is critical to engraftment, Radiology, IRCSS Casa Sollievo Della Sofferenza, San Giovanni TRM and OS. Therefore the use of double UCBT (dUCBT) Rotondo (FG), Italy rather than single unit is perceived as beneficial in adult recipients. The role of factors such as HLA disparity, Introduction: Bone marrow (BM) harvesting is universally total nucleated cell dose (TNC) and CD34 þ dose, which recognized as a safe procedure. There are few risks for donors produce optimal outcomes in adult UCBT, is still being and serious complications rarely occur. As with most invasive elucidated. Here we analyse immunologic and haemopoietic procedures, bleeding and infection are listed among the most factors affecting transplant outcome in 20 adult dUCBT common risks, while rarer complications have been described, demonstrating that CD34 þ count, and not TNC, impact on including air or fat embolism and fracture of the ilium. OS at one year. We report an uncommon case of pseudoaneurysm of the Materials (or patients) and methods: Between November superior gluteal artery causing gluteal compartment syndrome 2009 and January 2014, 20 patients underwent dUCBT at two as a complication of bone marrow harvesting. transplant centres for malignant haematological disorders Materials (or patients) and methods: A 52-year-old female (AML 35%, ALL 15%, CML 15% and 35% other diseases). The underwent BM harvest serving as the allogeneic haploidentical selected units used were HLA matched for 4/6 (24 units) or 5/6 donor for her daughter suffering from recurrent Hodgkin (16 units) at low resolution for HLA-A and B and a minimum of lymphoma after autologous bone marrow transplantation. The one allele level match for HLA-DRB1. Units were selected to harvesting was taken from the bilateral iliac crests under provide a combined CD34 þ cell dose in excess of 2 105/kg, general anesthesia using a standard technique. The patient preferably with no single unit providing less than 1 105/kg. reported severe pain in the left buttock at the time without The cohort had either the Minnesota RIC regimen (Flu 40 mg/ paresthesia or functional weakness of the ipsilateral lower m2, Cyclo 50 mg/kg and TBI 2 Gy) or the MAC regimen (Flu limb. At the time physical examination showed no evidence of 25 mg/m2, Cyclo 60 mg/kg and TBI 13.2 or 14.4 Gy). Both mass in her buttock, no abnormal neurological findings groups received CyA and MMF for GvHD prophylaxis. elsewhere, particularly in the left lower limb. Initially the Results: Median age at transplant was 44.2yrs (range 18-65). patient was treated with analgesia and observation. Progres- The median time from initiation of search to transplant was sively marked bruising developed over the left buttock and 192 days (range 102-363). The HLA disparity of units selected thigh, and functional impotence of the left lower limb for each patient, once high resolution typing was performed gradually appeared. for HLA-A, B, C, DRB1 and DQB1, varied from 4/10 to 8/10. Results: A computed tomography (CT) scan revealed a The main factors contributing to the level of disparity nonspecific heterogeneous tissue mass measuring 8.7 x 3.5 were variation in HLA-B/C and HLA-DRB1/DQB1 associations x5.3 cm into the left gluteus maximus muscle and a 1.5 cm as well as allele variation at HLA-B. The median TNC from area of active extravasation of contrast within the posterior each unit was 2.98 107/kg (range 1.19-6.55) and the part of the same muscle. We discussed this case with a median CD34 þ cell dose from each unit was 2.01 105/kg Consultant Interventional Radiologist and a formal angiogram (range 0.9-4.88). The average time to neutrophil and platelet was performed with a view to embolization. Selective left iliac engraftment was 21 and 34 days respectively (ranges being 9- artery angiogram demonstrated a large gluteal superior artery 36 days and 12-54 days). Three patients died prior to platelet pseudoaneurysm. Embolization of the superior gluteal artery engraftment; one death was due to relapse, one aGvHD was performed with Glubran 2 acrylic glue (GEM, Viareggio, involving skin and liver and the third aGvHD of the gut. One Italy), and a follow-up angiogram demonstrated no residual year OS was 75%. Mild aGvHD involving the skin alone (up to filling of the pseudoaneurysm. After the successful glue grade II) occurred in 15 patients. There were 6 cases of gut embolization of the right superior gluteal artery, the patient GvHD ranging from grade II-IV and in 2 of these cases liver had immediate resolution of her pain. The lady was discharged GvHD also occurred. later with almost complete resolution of pain and no left lower Conclusion: Most published data emphasise the importance limb weakness. of HLA matching and TNC for cord unit selection in HSCT. In Conclusion: Our patient developed an uncommon complica- this cohort of patients, cord blood units were selected with tion of BM harvesting, namely a pseudoaneurysm of the left emphasis on CD34 þ content as well as HLA match. Engraft- gluteal artery, which caused severe neuropathic pain and ment occurred in an appropriate time frame for the majority of weakness most likely linked to sciatic nerve compression. cases. 75% of patients developed limited GvHD of the skin Nevertheless, once the diagnosis was made and she under- indicating an allogeneic response. This may be supportive of went percutaneus embolization, pain relief occurred promptly the presence of a GvL effect. Our data shows no difference in and was almost durable. It is important to rule out a OS with respect to HLA matching.

S566 Whilst all selected cord unit pairs fulfilled national guidelines specific transplant protocols a mononuclear cell collection for TNC, there was no correlation between TNC dose and OS. from peripheral blood (for DLI) may be required. However there was superior OS for patients receiving a Before mononuclear cell donation, a complete work-up is combined CD34 þ cell dose above 2.8 105/kg (Po0.01). performed, follwing the applicable laws and procedures, These results indicate the need to prioritise a high CD34 þ evaluating donor suitability and determining any infectious dose when selecting units for dUCBT. diseases transmissible by lymphocyte donation. In order to Disclosure of Interest: None declared. verify lymphocyte collection safety, donors are evaluated for lymphocytes typing: CD3 þ , CD4 þ , CD8 þ , CD19 þ , CD56 þ , before mononuclear cell collection and after donation (within 1-3 months after donation). Following the donation, the well-being of the donor must be ascertained Stem cell donor by the DC physician until at least 6 months after donation, or in case of any abnormal ailments, until the donor is free of ailments. AB032 Materials (or patients) and methods: We evaluated all the The role of HLA high resolution typing in umbilical cord requests of second or further lymphocyte donation, analyzing blood transplantation the experience of Italian Registry and the incidence of a second or further lymphocyte donation request following HSC A. Sun1,*, S. Yang1,J.He1,D.Wu1 1 donation. The First Affiliated Hospital of Soochow University, Jiangsu Results: From 1989 to 2013, first HSC products donated by Institute of Hematology, Suzhou, China Italian donors were 2999 (2160 BM and 839 PBSC): 5% of these donors are asked for a ‘‘second’’ donation: of these 55% were Introduction: To analyze HLA matching (low or high DLI. Lymphocyte donation were more frequent after PBSC resolution) on outcomes of umbilical cord blood trans- donation: for these donors the incidence of a ‘‘second’’ plantation (UCB) and the value of clinical application. donation is 8,3% and on 87% of cases were for mononuclear Materials (or patients) and methods: From October 2010 to cell. After bone marrow donation the incidence of lymphocyte July 2012,26 patients were selected umbilical cord blood donation were 22% transplantation in our hospital,in the absence of a suitable HLA- Analyzing CD3/CD4/CD8 typing pre and post lymphocyte identical sibling donor or alternative donors.We examined the donation, we observe that in 12 donors more than one request effects of HLA matching (low or high resolution) on engraft- for lymphocyte donation were made by Transplant centers. On ment of leading, engraftment,graft-versus-host disease (GVHD), 30% of these donors CD8 count were persistently low after infection and mortality.Performed mainly by sequence-specific donation. oligonucleotide probe (SSOP) methods, sequence-based typing Conclusion: Analyzing Italian Bone Marrow Donor Registry (SBT) methods and high resolution sequence-specific pri- experience, after HSC donation, the incidence of a ‘‘second’’ mers(SSP) methods,retrospective HLA matching of cord blood,- donation request is 5%, more after PBSC donation than after typed at high resolution for HLA-A,-B, -C, -DRB1, -DQ, and -DP BM donation (8,3% vs 2,7%); Lymphocyte donation were more and low resolution for HLA-A,B,DRB1. frequent after PBSC donation: for these donors the incidence Results: Compared single UBC units transplantation with of a ‘‘second’’ donation is 8,3% and on 87% of cases were for double UBC units,the hematopoietic reconstitution rate was mononuclear cell. For donors asked for more than one 88% vs 56%.The median total nucleated cells(TNC) of single Z lymphocyte donation, CD3/CD8 count have to be controlled and double cord blood was 5.5 107/kg, when TNC 5 107/ and monitored because frequently this value, in our experi- kg could promote neutrophil recoveries (Po0.05). The UBC ence could be persistently low on 30% of donors. units were a 6/10 HLA high resolution match or better with Disclosure of Interest: None declared. each other could promote UCB implantation advantaged (Po0.05), accelerate platelet recovery (Po0.05) and reduce more risk of acute GVHD (grade III -IV) (Po0.05). There are AB034 different between the group 5-6/6 HLA low resolution and 3-4/ Retrospective analysis of peripheral stem cell apheresis 6.but no statistical difference (P40.05). from allogeneic donors: single center experience Conclusion: The HLA High resolution typing can select the M. Tenorio1,*, G. Moreno1, A. Jime´nez1, P. Herrera1,J.Lo´pez better UCB than HLA low-resoulution. It has important clinical Jime´nez1 value on promoting hematopoietic reconstitution and redu- 1Haematology and Haemotherapy, HOSPITAL RAMON Y CAJAL, cing complications after UCB transplation. MADRID, Spain Disclosure of Interest: None declared. Introduction: As the repertoire of allogeneic donors widens with the introduction of haploidentical stem cell transplanta- AB033 tion and older aged donors, the possibility to encounter bad Lymphocyte donations as a further donation of mobilizers increases and new approaches and complications hematopoietic stem cell volunteer donors: the experience may be encountered during apheresis. of the Italian Bone Marrow Donor Registry (IBMDR) Materials (or patients) and methods: We retrospectively A. Vassanelli1,*, N. Sacchi2, S. Pollichieni2 analyzed the data collected during apheresis of peripheral 1Transfusion Medicine, Azienda Ospedaliera Universitaria stem cell donors at Ramon y Cajal hospital from January 2012 Integrata Verona, Verona, 2IBMDR, Ospedale Galliera, Genova, to November 2014 to establish the efficacy of our mobilization Italy protocol and the safety of our leucoapheresis stem cell collection method. Our standard procedure includes mobiliza- Introduction: The Italian Bone Marrow Donor Registry was tion using 5 m/Kg/12 hours of filgrastim sc during five days; established in Italy in 1989, with the role of providing an apheresis is then started on day 5 and continued the following unrelated volunteer to haematological patients waiting for a day (with same dose G-CSF) if objective CD34 þ /Kg is not transplant and who do not have the ideal donor (an identical reached. Three volemias are processed and stem cells are sibling). collected using COBE-SPECTRA and ACD-A as anticoagulant A Haematopoietic Stem Cell donation can be carried out from (proportion of 1:20 and rate infusion 0.8 mL/min/L).Data both bone marrow and peripheral blood after administration collected included: age, sex, type of donor, type of venous of G-CSF. In particular, the prospective donor must be access, CD34 þ /microL (peripheral blood), CD34 þ /Kg (apher- informed - that, in some circumstances, the donor may be esis product) and complications associated with apheresis asked an additional donation for the same patient. Following technique.

S567 Results: All 64 allogeneic stem cell donors (35 related Materials (or patients) and methods: We report a HLA-identical, 10 unrelated HLA-identical, 18 haplo- 22-year old male with severe SCD-/-Thalassemia and identical, 1 auxiliary for secuential transplantation) submitted transfusion related HLA alloimmunization who received a to leucoapheresis stem cell collection achieved objective bone marrow alloHSCT from his haploidentical father. In CD34 þ /Kg (between 3-4CD34 þ /Kg). The results were: addition to normal organ function testing, the pre-transplant ratio female/male 1:2, mean age 46 years-old (median 47 work-up included HLA antibody screening. To reduce the risk years-old; range 20-73 years-old), main venous access of graft rejection, the preparative regimen included in a was peripheral (94%), mean volemia processed was pre-transplant immunosuppressive therapy phase (modified 14608 mL, mean CD34 þ /mL was 76 (range 9-230) and from Anurathapan U, BBMT 19:1259, 2013), using two four day mean CD34 þ /Kg was 7.92. Patients who achieved courses of fludarabine (Flu) 40 mg/m2/d and dexamethasone o3x106 CD34 þ /Kg in one single apheresis session presented 40 mg/d, six and three weeks before the start of the RTC CD34 þ /mL between 9 and 37; and those with 43x106 conditioning with antithymocyte globuline (ATG, rabbit) CD34 þ /Kg between 25 and 236. Four patients underwent less 1.5 mg/kg on days -12 to -10; thiotepa 5 mg/kg on day -9; than 3 volemias: 3 were shortened due to complications Flu 40 mg/m2/day, each dose followed by IV busulfan 100 mg/ related to the apheresis (2 paresthesias associated with m2 on days -6 to -3. Graft versus host disease (GVHD) hypocalcemia and 1 dizziness) and 1 due to an extremely prophylaxis done with posttransplantation cyclophosphamide high count of CD34 þ /microL in peripheral blood. Five (Cy) 50 mg/kg on day þ 3 and þ 4; cyclosporine A and patients required a second apheresis to achieve objective mycofenolate mofetil were started on day þ 5. One CD34 þ /Kg and 1 patient needed a third session; all were cycle of red blood cell exchange was performed before the more than 50 years-old except one patient which presented a RTC conditioning and plasmapheresis was done on days -13 significant weight difference with the recipient. Two patients and -1. Disease response was measured by HbS levels received off-label plerixafor at standard dose (240 mg/Kg sc) using weekly hemoglobin electrophoresis. Monthly chimerism 10 hours before the apheresis due to insufficient collection of studies by PCR analysis were done on peripheral blood stem cells. One of them was a 49 year-old woman with no CD3 þ cells. known comorbidity except positive anti-HCV and the other Results: Time to neutrophil and platelet recovery over was a 63 year-old man with no significant history. Both 500 103/mm3 and 50 103/mm3 were 29 and 43 days increased their peripheral blood CD34 þ count after respectively; transfusion independency was reached by day plerixafor þ filgrastim (from 9 to 44.5 CD34 þ /mL and 9 to 33 þ 36. Initial donor chimerism was mixed at 20% and the trend CD34 þ /mL respectively) and both accordingly increased is increasing; at time of last follow-up, donor cells dominated CD34 þ /Kg collected (from 0.7 to 2.86 CD34 þ /Kg and 0.53 at 86% while immunosuppression is still ongoing. HbS levels to 3 CD34 þ /Kg). (baseline 68%) continuously decreased after start of the Conclusion: New strategies have to be adopted in preparative regimen too1% on day þ 77. Donor specific potential bad mobilizers in allogeneic donors. The allo-antibodies (DSA) were significantly reduced by the PTIS addition of plerixafor to G-CSF may be a possible solution. treatment and the subsequent alloHSCT to where there are Appropriate prospective studies need to validate this presently undetectable. With a follow up of 257 days, the pt approach. has not developed any GVHD or serious early TRT. References: Hauge AW, Haastrup EK, Sengelev H, Minulescu L, Conclusion: We suggest that our approach with a modified Dickmeiss E, Fischer-Nielsen A. Addition of plerixafor for pre-conditioning immunosuppressive phase followed by a RTC CD34 þ cell mobilization in six healthy stem cell donors preparative regimen and posttransplantation Cy provides ensured satisfactory grafts for transplantation. Transfusion sufficient T-cell depletion and tolerance induction to allow 2014; 54 (4): 1055-8. durable donor cell engraftment and SCD control in adults. The Disclosure of Interest: None declared. presence of DSA should not be a barrier to HLA haploidentical related alloHSCT. This novel haplo-transplant program war- rants further study in a trial of SCD in selected patients. AB035 Disclosure of Interest: None declared. Successfull engraftment and clearance of donor specific antibodies after haploidentical related stem cell transplantation for an adult patient with sickle cell disease S. Fu¨rst1,*, E. Bernit2, J. El Cheikh1, A. Granata1, S. Harbi1,B. Graft-versus-host disease – clinical Mohty1, L. Caymaris1, C. Lemarie3, B. Calmels3, C. Chabannon3,A. Basire4, C. Picard4, P. Poullin5, B. Andersson6, D. Blaise1 1Unite´ de Greffe, Institut Paoli-Calmettes, 2Medecine Interne, Hopital La Conception, 3Therapie Cellulaire, Institut Paoli- AB036 Calmettes, 4HLA, EFS, 5Hematologie, Hopital La Conception, Reduction in the Incidence and Severity of Graft Versus Marseille, France, 6Stem Cell Transplantation, MD Anderson, Host Disease in HLA-Matched and Haploidentical Bone Houston, United States Marrow Transplantation Using Post-trasplant Cyclophosphamide: a Multicentric Experience in Mexico R. Ovilla Martı´nez1, A. Cabrera Garcia2,*, A. OLAYA3, U. PEREZ4, Introduction: Sickle cell disease (SCD) is one of the most 5 2 2 2 common genetic disorders in the world and it remains a A. LIMON , P. WEBER , D. CALLES , L. REYNOLDS on behalf of disease with high risk of morbidity and mortality. Allogeneic GRUPO MULTICENTRICO DE TRASPLANTE HAPLOIDENTICO 1HOSPITAL ANGELES LOMAS, HUIXQUILUCAN, Mexico, hematopoietic stem cell transplantation (alloHSCT) is the only 2 curative treatment modality. Reports of adults undergoing HEMATOLOGY, HOSPITAL ANGELES LOMAS, HUIXQUILUCAN, 3BONE MARROW TRANSPLANT, INSTITUTO NACIONAL DE successful alloHSCT are limited because of a lack of suitable 4 donors and high treatment related toxicity (TRT) after PEDIATRIA, MEXICO CITY, BONE MARROW TRANSPLANTATION, INSTITUTO MEXICANO DEL SEGURO SOCIAL. UMAE HE. GRAL DIV. conventional myeloablative conditioning regimen. Early 5 reports of non- myeloablative conditioning regimens were MANUEL AVILA CAMACHO, BONE MARROW TRANSPLANTATION, disappointing because of a high incidence of graft failure (GF) INSTITUTO MEXICANO DEL SEGURO SOCIAL. UMAE HE. GRAL DIV. and disease recurrence. The recent development of reduced MANUEL AVILA CAMACHO, PUEBLA, Mexico toxicity conditioning regimens (RTC) with lower TRT and use of haploidentical family donors has widened the applicability of Introduction: The Graft Versus Host Disease (GVHD) grades alloHSCT. Unfortunately, most SCD patients (pt) are alloimmu- II-IV is present in up to 40% in Human Leucocyte Antigen nized which further compromises HLA mismatched cell (HLA) - identical related transplants and up to 80% of non- engraftment. related. Particulary the HLA-haploidentical bone marrow

S568 transplantation (BMT) has been associated with significant Materials (or patients) and methods: The primary endpoint risks of graft rejection and severe GVHD, high dose of of the study is the reduction in cGVHD incidence in patients Cyclofosfamide (Cy) after BMT inhibits both graft rejection (pts) subjected to prophylactic ECP. Secondary endpoints and GVHD. We want to share our experience in preventing are delayed appearance of cGVHD, a reduced incidence of acute GVHD (aGVHD) and chronic GVHD (cGVHD) in HLA - cGVHD complications, and a lower steroid dose compared to haploidentical alloBMT and HLA-matched related BMT using control population. All the pts who underwent peripheral Cy post-Transplant strategy. T-repleted HSCT with conditioning regimen Thiotepa Materials (or patients) and methods: Thirty seven patients 5 mg/kg, Fludarabine 90 mg/mq and Melphalan 100 mg/mq from three Mexican Institutions, all of them were in advanced are eligible. aGVHD prophylaxis is Methotrexate and stages of malignant and bening hematological diseases. In the cyclosporine, that is tapered by day þ 90 with a weekly group of malignant diseases (83.8%), was performed HLA- dose reduction of 10%. Prophylactic ECP is started when haploidentical alloBMT in 23 patients and HLA-matched related cyclosporine levels in the bloodstream is below 100 ng/ml alloBMT in 8 patients. In the group of benign diseases (16.2%), (fig1). All pts have to be in complete remission (CR) at the time was performed HLA - haploidentical alloBMT in 6 patients. of ECP start, with a performance status 0-2 according to ECOG Results: The 56.7% (21) were male, mean age 26 years (range scale and an age of 18-65 years. Control population is 2-67 years), an average follow-up of 235 days (range 14-719 represented by a cohort of pts with the same clinical days). All patients Received non-myeloablative conditioning characteristics, treated in the same years, with the same and solely Cy 50 mg / kg on days 3 and 4 after transplantation conditioning regimen, graft, aGVHD prophylaxis and that ar in for patients related HLA-matched BMT as GVHD prophylaxis; CR at 100 days after HSCT. for HLA-haploidentical BMT, Mycophenolate mofetil (from day Results: From May 2009 to November 2014 18 pts were 5 to 35) and Tacrolimus (from day 5 to 180) was added. The enrolled; clinical characteristics of ECP and control group (48 incidences of aGVHD grades I through II and grades III through pts) are summarized in Table 1. Three of 18 pts are not IV were 40.5% and 19% respectively. The GVHD-related included in our analysis because of early relapse. Five pts out mortality was 5.4%. cGVHD grade I through II occurred in 3 of 15 completed the study, 3/15 pts are ongoing without patients, only one patient died with grade IV. Transplant- evidence of GVHD, while 7/15 stopped ECP because of cGVHD related mortality was 37.8%; there were 8 relapses and 4 died. in 5 cases, consensus withdrawal in 1 case and death for an Were registered 10 graft failures (27%) of which 8 patients infectious complication in the last case. A median of 11 (range died. 4 out of 6 patients in the group of benign diseases had 4-20) procedures was performed, without any complication no graft and they required a second BMT. The overall survival due to ECP. In ECP group we observed cGVHD in 9715 pts (OS) at day 180 by type of transplant, HLA-haploidentical BMT (60%), after a median time of 11 (7-21) months from HSCT, in malignant diseases, HLA-matched related BMT and HLA- with a grading of severe in 1 (6%), moderate in 6 (40%) and haploidentical BMT in benign diseases was 56%, 62% and 66% mild in 2 (13%). GVHD involved skin or mucosa in 7 pts, liver in respectively. As a complication related to Cy therapy, 1 and kidney in 2 (nephrotic syndrome). All 7 (47%) pts with a hemorrhagic cystitis was Observed in 32% (8 patients). moderate/severe GVHD received steroid. In control population Conclusion: Post-transplantation Cy is able to reduce the we recorded 25/48 cases of cGVHD (52%): severe in 9 cases developing chronic GVHD and reduces the severity of acute (18%), moderate in 11 (22%) and mild in 5 (10%). The median GVHD, it plays an important role in the feasibility of a time of appearance was 8 months (r 4-41). Nineteen (40%) pts haploidentical transplant because the lower incidence of received steroid therapy. GVHD, however in bening diseases we had high rates of graft failure. In Mexico there is a little experience about this Table 1 extrategy. Our results showed one of the most satisfactory and encouraging studies on the use of post-transplantation Cy performed in Latin America. ECP CONTROL n18 n48 References: 1. Luznik L, Fuchs EJ, et al. Post-transplantation high-dose cyclopyhosphamide is effective single agent GVHD Sex M/F 10 / 8 23 / 25 prophylaxis that permits prompt immune recontitution after Age median (range) 61 (48-67) 58 (25- 66) myeloablative HLA matched related and unrelated bone Diagnosis marrow transplantation. Biol Blood Marrow Transplant AML/MDS 14 (78%) 29 (60%) ALL — 2 (4%) 2007;13(2Suppl):4. HD and NHL 4 (22%) 8 (17%) 2. Bolan˜ os-Meade J, Fuchs EJ, et al.: HLA-haploidentical bone MM — 9 (19%) marrow transplantation with posttrasplant cyclophosphamide DONOR explands the donor pool for patients with sickle cell disease. Sibling/MUD 9 / 9 (50 / 50%) 31 / 17 (65 / 35%) Blood 120:4284-4292, 2012 8/8 HLA matched 14 38 Sex M/F 12 / 6 34 / 14 Disclosure of Interest: None declared. Median follow up months (range) 19 (6–65) 82 (9–125) AB037 Prospective clinical study on extracorporeal photopheresis (ECP) as prophylaxis for chronic Graft Versus Host Disease (cGVHD) prophylaxis: an interim analysis E. Zucchetti1,*, G. Grillo1, G. Bertani2, M. L. Pioltelli1, B. Forno1, E. Ravano1, I. Lotesoriere1, P. Marenco1, R. Cairoli1 1Department of Hematology, 2Department of Transfusional Medicine, NIGUARDA Hospital, Milan, Italy

Introduction: GVHD is the most frequent cause of Non Relapse Mortality and morbility after hematopoietic stem cells transplantation (HSCT). For prophylaxis of acute GVHD (aGVHD) calcineurin inhibitors and short term Methotrexate are the gold standard, while there are no evidences about prophylaxis of cGVHD. ECP emerged in the last ten years as a second line therapy for GVHD, but its role as prophylaxis for cGVHD is still debated. We started a prospective clinical study to evaluate the role of ECP in cGVHD prophylaxis.

S569 Conclusion: We observe the same cGVHD incidence in correlation with these factors, detection of aGvHD risk factors both treatment and control group; however in the ECP group can help increase the success of allo-HSCT. there is a trend versus fewer severe grading of GVHD Disclosure of Interest: None declared. and versus a more delayed onset median time (p ns). Interim analysis of ECP as cGVHD prophylaxis show this AB039 protocol is feasible and safe, so we are comfortable to Rituximab treatment in patient with steroid-refractory continue the study. chronic GVHD: a single-center experience Disclosure of Interest: None declared. I. Donnini1,*, C. Nozzoli1, A. Gozzini1, R. Saccardi1, S. Guidi1, A. Bosi2 AB038 1Bone Marrow Transplantation Unit, 2Hematology department, Evaluation Of Acute Graft Versus Host Disease In AOU Careggi, Florence, Italy Allogeneic Hematopoietic Stem Cell Transplantations F. Sahin1,*, M. Comert1, A. Uysal1, M. Tombuloglu1, N. Soyer1, Introduction: Chronic GVHD (cGVHD) is the most G. Saydam1, F. Vural1 important determinant of late morbidity and mortality in 1hematology, Ege University Medical Faculty, IZMIR, Turkey long-term survivors after allogeneic stem cell trans- plantation. Patients (pts) with cGVHD steroid-refractory Introduction: Acute Graft-Versus-Host-Disease (aGvHD) is a have a poor outcome. Several authors reported promising major cause of morbidity and mortality that occurs in first 100 findings with the use of rituximab for salvage therapy of days after allogeneic hematopoietic stem cell transplantation cGVHD.1 (allo-HSCT). The incidence of aGvHD in full-matched allo-HSCT Materials (or patients) and methods: Our retrospective is 30-50%. We evaluated the incidence and risk factors for study aimed to analyze the results of rituximab salvage aGvHD in patients treated with allo-HSCT. therapy in a series of 23 consecutive pts with refractory cGVHD Materials (or patients) and methods: Seventy four patients treated between 2008 and 2014 in our center. Pts, transplant, who were transplanted in our unit in January 2011 and June graft and GVHD characteristics are summarized in Table 1. For 2014 were analyzed retrospectively. GVHD prophylaxis, pts received either CsA alone (n ¼ 1; 4.3%) Results: Patients’ and donors’ characteristics were showed or CsA and mycophenolate mofetil (MMF, n ¼ 4; 17%), or CsA in table 1. All of the patients received cyclosporine and and MTX (n ¼ 18; 78%). Acute and cGVHD were evaluated methotrexate for GVHD prophylaxis. The rate of AHSCT with according to the NIH standard criteria. Median age same patient and donor blood group was 44.6% (n ¼ 33) and was 42 years (range 16–60) with 56.5% males, and 65% of aGvHD was seen 36.6% (n ¼ 12) in this group. The pts having a myeloid malignancy. At the time of rituximab, all rate of aGvHD in different patient and donor blood groups pts presented with extensive and refractory cGVHD; in was 43.9% (n ¼ 18). There was no significant difference in particular, six pts showed autoimmune cytopenias such as aGvHD between two groups. But; 7 of the 9 patients autoimmune hemolytic anemia (AIHA), autoimmune throm- with Rh incompatibility showed aGVHD (P ¼ 0.015). The bocytopenia (AIT) and acquired hemophilia A (AHA). Pts recipient was male and the donor was female in 24 AHSCT received and failed (in addition to corticosteroids) several lines and 10 (41.6%) of those, we detected aGvHD. The recipient of prior immunosuppressive therapy: CsA (n ¼ 7), MMF was female and the donor was male in 13 AHSCT and 4 (n ¼ 12), imatinib (n ¼ 3), ECP therapy (n ¼ 8), low-dose MTX (30.7%) aGvHDs were seen in this group. There was no (n ¼ 1), sirolimus (n ¼ 1) or fk506 (n ¼ 1). The median time significant difference between groups in terms of develop- between cGVHD diagnosis and rituximab therapy was 2 ment of aGvHD (P40.05). (range, 0.5–51) months. Rituximab was given at dose of 375 mg/m2/week for 4 consecutive weeks. All pts completed the four courses treatment and 9/23 (39%) received subsequent maintenance therapy (one rituximab infusion per All patients aGVHD No signs of p n(%) n(%) aGVHD n (%) month). Results: We observed a 13/23 (56.5%) overall response rate. # of patients 73 (100) 37 (50.6) 36 ( 49.4) cGVHD responses were as follows: 6 out of 14 skin, 1 out of 3 Sex of patients (F/M) 27/46 () 13/ 24 14/22 P40.05 oral mucosa, 3 out of 6 liver, 3 out of 3 AIHA, 1 out of 3 Diseases 3 1 2 P40.05 articular, 1 out of 2 (AIT), and 1 out of 1 AHA responses. AML 9 7 2 17/23 pts (74%) experienced cGVHD relapse. At 24 months of ALL 1 7 6 AA 3 1 4 median follow-up, ten patients (43.4%) had died; five (22%) MM 5 3 1 MDS 5 3 1 CML 4 2 0 MMF 2 1 1 Lymphoma 23 3 0 Conditioning regimen 44 20 24 P40.05 Myeloablative 29 17 12 Non-myeloablative HLA Full matched 69 35 34 9/10 matched 2 1 1 Haploidentic 2 1 1 Median age 47 (17- 67) 45 (17- 67) 45 (18- 65) P ¼ 0.341 Median donor age 42.5 (11- 64) 45 (13- 64) 40 (11-64) P ¼ 0.138 CMV DNA positive 13 (17.6) 5 (38.4) 8 (61.6) P40.05

AML: acute myeloid leukemia ALL: acute lymphoblastic leukemia AA: aplastic anemia, MM: multiple myeloma MDS: myelodysplastic syndrom, CML: chronic myeloid leukemia MMF: myelofibrosis Conclusion: The risk factors for aGvHD development in allo- HSCT was reported as advanced donor and recipient age, CMV positivity, male recipient and female donor, HLA mismatch, type of GvHD prophylaxis, the intensity of conditioning regimen. Even though we didn’t find any significant

S570 due to infections and three (13%) due to cGVHD. No toxicity occurring 21 months post-transplantation. Among the 64 related to rituximab was observed. patients, the serum creatinine levels increased two-fold or more during the first month after transplantation only in 3 Conclusion: The pathophysiology of cGVHD is still poorly patients (4.7%). With a median follow-up of 37 months, the understood. As already observed in some autoimmune estimated probabilities of disease free survival (DFS) and disorders, the beneficial effect of B-cell depletion with overall survival (OS) were 65.6±6.1% and 67.2±5.9%, rituximab highlights the potential primary pathologic role of respectively (Figure 3). B lymphocytes in the development of cGVHD.2 In the light of Conclusion: This retrospective analysis suggests that this altered B-cell homeostasis in cGVHD, the role of rituximab stepwise administration of tacrolimus for GVHD prophylaxis in controlling the clinical manifestations of cGVHD has been in patients undergoing HLA-haploidentical HSCT is associated studied.3 In our study, an overall response rate of 56.5% was with high engraftment rates, a low incidence of aGVHD, and observed, especially at the skin, liver and autoimmune excellent NRM, OS and DFS. Future strategies to decrease the cytopenias. In summary, this series suggest that rituximab incidence of cGVHD while preserving antitumor activity are used as salvage therapy for refractory cGVHD has a promising necessary. efficacy and an acceptable safety profiles. Disclosure of Interest: None declared. References: 1. Kharfan- Dabaja MA, Mhaskar AR et al. Efficacy of rituximab in the setting of steroid-refractory chronic graft- AB041 versus-host-disease: a systematic review and meta-analysis. Extracorporeal photopheresis variably affects Biol Blood Marrow Transplant 2009; 15: 1005-1013. profile in patients with chronic Graft Versus Host Disease, 2. Zaja F, Bacigalupo A, et al. Treatment of refractory chronic showing possible prognostic implications GVHD with rituximab: a GITMO study. Bone Marrow Transplant C. G. Valentini1, N. Orlando1, G. Massini1, N. Piccirillo1, 2007; 40: 273-277. 1 2 1 1 2 3. Kim SJ, Won JH. B cell homeostasis and the develop- M. Maresca , E. Metafuni , R. Putzulu , M. Bianchi , P. Chiusolo , S. Sica2, L. Teofili1,* ment of chronic graft-versus-host-disease: implications 1 2 for B cell-depleting therapy. Leuk Lymphoma 2012; 53: Transfusion Medicine, Hematology Department, Catholic 19-25. University, Rome, Italy Disclosure of Interest: None declared. Introduction: Molecular and cellular mechanisms underlying the immunomodulatory effect of of extracorporeal photo- AB040 pheresis (ECP) in Graft Versus Host Disease (GVHD) are not Feasibility of stepwise administration of tacrolimus as a completely understood. We investigated the cytokine profile graft-versus-host disease prophylaxis in non-T-cell- in patients with chronic GVHD (cGVHD)before and after ECP. depleted haploidentical hematopoietic stem cell Materials (or patients) and methods: Plasma samples transplantation for patients with hematological were collected immediately before (day 1) and 24 hours malignancies after ECP (day 2) in 12 pts (M/F 9/3; median age 51 years, L. Gao1,*, J. Liu1, Y. Zhang2, X. Chen1, L. Gao1, C. Zhang1, Y. Liu1, range 19-65) with cGVHD undergoing two consecutive ECP P. Kong1, Q. Wang1, A. Sun1,X.Du1, D. Zeng1,J.Li1, H. Liu1, procedures every month. Diagnosis were: AML (7 pts), MDS (2 X. Peng1, X. Zhang1 pts), MPN ( 2 pts) and ALL (1 pt). At the time of sample 1Department of Hematology, Xinqiao Hospital, Third collection, all pts were in complete remission. Immunosup- Military Medical University, Chongqing, China, 2Department of pressive therapy consisted of cyclosporine (8 pts), prednisone Health Statistics, College of Military Preventive Medicine, (7), mycophenolate mofetil (4), alone or in combination. In 7 Third Military Medical University, Chongqing, China, chongqing, pts GVHD was severe and involved skin (6 pts), liver (3), China gastrointestinal tract (2), musculoskeletal system (3), lung (1), eyes (1). In the other 5 pts the cGVHD was moderate and Introduction: Graft-versus-host disease (GVHD) is the primary involved liver (3 pts), skin (3), eyes (2), gastrointestinal tract (1) complication of HLA-haploidentical hematopoietic stem and musculoskeletal system (1). Multiplex analysis of 27 cell transplantation (HSCT). To date, the ideal prophylaxis was performed using the Luminex system on a regimen, which minimizes GVHD but does not abrogate Bioplex (Bio-Rad) cytomer (Bio-plex Human Cytokine 27-Plex the graft-versus-tumor (GVT) response and delay post- Panel). Plasma samples from 10 healthy blood donors were transplantation immune recovery, has not been described. used as controls. However, there is little information regarding the correlations Results: The pts had similar levels of baseline cytokines as of aGVHD incidence and tacrolimus blood concentration at controls, except for lower concentrations of IL2 (P ¼ 0.006), HLA-haploidentical HSCT. Our goal in this study was to IL17A (P ¼ 0.004), GM-CSF (P ¼ 0.050) and CCL5 (Po0.001). investigate the feasibility and clinical value of descending Among hematological parameters, platelet count was stepwise administration of tacrolimus on the prophylaxis of the most significant predictor for cytokine levels (Po0.05 in GVHD in patients who underwent HSCT from HLA-haploiden- 20/27 cytokines, with r between 0.06 and 0.70 at Spearman tical donors. Materials (or patients) and methods: Between June 2008 and October 2010, 64 consecutive patients with hematological malignancies who underwent HLA-haploidentical HSCT at Xinqiao Hospital were enrolled in this study. The conditioning regimen, infection prevention and supportive care were according to previous experience. The primary endpoint of the study was the cumulative incidences of aGVHD and cGVHD after transplantation. The secondary endpoints included engraftment rate, tacrolimus-associated toxicity, incidence of infection and relapse, disease-free survival (DFS) and overall survival (OS). Results: 63 of the 64 assessable patients achieved full donor chimerism by day 30 after HSCT. The 2-year cumulative incidences of cGVHD and extensive cGVHD were 46.0±6.3% and 9.0±3.8%, respectively (Figure 2b). Relapse occurred in 20 of 64 patients. The estimated cumulative incidence of relapse was 31.7±5.9%, with the latest event

S571 test). Similarly, the platelet content of the apheresis References: Harper, S. J., J. Moorhouse, et al. (1996). "The product significantly correlated with the cytokine levels after beneficial effects of oral nifedipine on cyclosporin-treated ECP. The concentration of all investigated cytokines and renal transplant recipients--a randomised prospective study." growth factors following ECP varied according to three Transpl Int 9(2): 115-25. different recurrent patterns: increase (3 pts), decrease (4 pts) Liesveld J Fau - Duerst, R., A. Duerst R Fau - Rapoport, et al. and no modifications (5 pts), respectively (group 1, 2 and 3 in "Continuous infusion cyclosporine and nifedipine to day Figure 1). In contrast, chemokine levels were scarcely þ 100 with short methotrexate and steroids as GVHD influenced by ECP. In two pts belonging to the group 2, prophylaxis in unrelated donor transplants." Bone Marrow data were confirmed in two different procedures. The first Transplant. 1999 Sep;24(5):511-6 pattern was basically observed in pts with extensive Morgano, A., I. Pierri, et al. (1990). "Decreased lymphocyte musculoskeletal involvement. Furthermore, 3 out of 5 pts blastogenesis, IL2 production and NK activity following belonging to the group 3 relapsed: (median overall survival of nifedipine administration to healthy humans." Eur J Clin 5,4 months for pts in group 3 versus undefined in groups 1 and Pharmacol 39(6): 545-50. 2, P ¼ 0.004, with a median follow from the study up of 5,3 and Solovera, J., M. Alvarez-Mon, et al. (1987). "Inhibition of human 9 months, respectively). natural killer (NK) activity by calcium channel modulators and Conclusion: Our data show that in pts with cGVHD the a calmodulin antagonist." The Journal of Immunology 139(3): cytokine profile varies after single ECP procedures: the 876-880. different pattern of modification can depend on disease-or Wang, D., K. Jiang, et al. "Determination of nifedipine in pt-specific clinical features and might have prognostic human plasma by ultra performance liquid chromatogra- implications. phyaˆh‘‘tandem mass spectrometry and its application in a Disclosure of Interest: None declared. pharmacokinetic study." Journal of Chromatography B 879(20): 1827-1832. AB042 Disclosure of Interest: None declared. Nifedipine and Ketoconazole is associated with a significant decrease in blood pressure and a loss of control AB043 of aGVHD and possibly GVL Successful use of therapeutic plasma exchange to N. G. Feldreich1,*, H. Kaipe1, O. Ringden1, B. Omazic2 overcome technical limitation of extracorporeal 1Terapuetic Immunology Laboratory Medicine, 2Oncology photopheresis in a case of severed hypertriglyceridemia & Pathology, Karolinska Institutet, Stockholm, Sweden and graft-versus-host disease R. D. C. TAVARES1,*, M. SILVA1, V. KUSSUMI1, I. MOTTA2, Introduction: The present study aims to investigate the A. SOUZA2, M. C. MOREIRA1, D. LERNER1, R. CASTRO1, immunomodulatory properties of nifedipine when accentuated L. F. BOUZAS1 in physiological effect by the combined treatment of 1Bone Marrow Transplant Unit, 2HEMOTHERAPY SERVICE, ketoconazole. Nifedipine, a antihypertensive calcium channel NATIONAL INSTITUTE OF CANCER, RIO DE JANEIRO, Brazil blocker has been investigated as an additional immuno- suppressant in both allogeneic hematopoietic stem cell Introduction: Extracorporeal photopheresis (ECP) is recom- transplantation (HSCT) and in renal transplantation [1,2]. While mended as second-line immunomodulatory treatment for Harper et al. 1996 could prove better initial transplant function steroid-refractory graft-versus-host disease (GVHD). Severe and decreased rejection in renal transplantation the in HSCT hypertriglyceridemia and hyperbilirubinemia may difficult were inconclusive. buffy coat separation during ECP. Therapeutic plasma In humans, a oral dose of 10 mg induces a decrease in exchange (TPE) is largely used in autoimmune diseases to blastogenesis and IL-2 production [3,4]. The effect is pro- lower the serum level of auto reactive antibodies and can also longed in NK-cells, whose cytotoxic process are susceptible to be used to lower a harmful level of lipids such as triglycerides. nifedipine [4,5]. Materials (or patients) and methods: A 42 year-old male Materials (or patients) and methods: After HSCT and patient underwent a second HLA-identical sibling bone during prophylaxis to control graft-versus-host disease marrow transplantation (BMT) for relapsed acute myeloid (aGVHD), 108 consecutive recipients were retrospectively leukemia. The conditioning regimen consisted of cyclopho- investigated for concomitant nifedipine and ketokonazole sphamide and total body irradiation. CSA and short-term (NA) treatment. The decrease in blood pressure by NA was methotrexate was used for GVHD prophylaxis and ursodeoxy- compared to the blood pressure decrease of nifedipine and cholic acid for sinusoidal obstruction syndrome prophylaxis. nystatin. The immunomodulatory effect of NA was investigated Laboratory values on day 0 revealed hypertriglyceridemia by whether steroid treatment for a suspected aGVHD reaction (464 mg/dL) and hypercholesterolemia (279 mg/dL). By day was initiated or not. þ 18 he developed grade III skin and biopsy-proven GI tract Results: NA significantly lowered the blood pressure in acute GVHD initially responsive to steroid therapy. He was comparison to nifedipine and nystatin with a decrease of discharged by day 22. Despite pharmacological (simvastatin, 19% vs. 6% in mean arterial pressure, P ¼ 0.02. NA evoked an fenofibrate) and nutritional treatments, cholesterol and inflammatory reaction in 9/10 cases compared to controls 46/ triglycerides levels increased progressively leading to renal 108, HR 3.33 [1.60-6.95] 95% CI. The majority of the NA group dysfunction, early CSA discontinuation and deep-venous 70% had cutaneous involvement and in the NA group 50% (5 thrombosis on his left leg. By day þ 84 his triglycerides levels out of 10) had only cutaneous involvement while 23% (22 out were 1112 mg/dL and he developed diffuse lichen-planus like of 98) in the control group had an inflammatory reaction skin lesions and hyperbilirubinemia with 13 mg/dL of total confined to the skin, P ¼ 0.06. The inflammatory reaction bilirubin (TB). The steroid dose was increased and tacrolimus seemed not related to graft-versus-leukemia, since three out of was started. After two weeks without response, MMF was four of the patients with less than ten days between NA and added resulting in a 40% drop of TB levels. By day þ 147 the inflammatory reaction relapsed. triglycerides levels increased up to 1675 mg/dL, gamma-GT Conclusion: Nifedipine administered to patients with ongoing and TB levels were 3251 and 7.7 mg/dL respectively. A double- azole treatment results in an increased physiologic effect lumen central catheter was inserted and ECP was unsuccess- and elicits an inflammatory reaction after HSCT. The clinical fully tried due to severe lipemia. TPE was performed in two reaction may be related to a loss of control over certain cell consecutive days without complications. The volume treated populations, possibly through decreased NK-cell cytotoxicity, was one total plasma volume and replacement fluid consisted since three out of four with a close relationship between NA of human albumin and fresh frozen plasma. and the aGVHD-like reaction later suffered a relapse in Results: The triglycerides levels decreased to 570 mg/dL in malignant disease. less than 24 hours. ECP could be started after two TPE

S572 procedures and was performed in three consecutive days. transplantation, and causes late-onset HC. The management of After two days, laboratory exams showed 469 mg/dL triglycer- BKV cystitis and nephritis sometimes may be very difficult and ides, 2039 mg/dL gamma-GT and 5 mg/dL TB. Bilirubinemia refractory all treatments. We presented our experience of BKV progressively decreased to 1.3 mg/dL, triglycerides levels infection, and management in transplanted patients in our remained stable and an improvement in skin lesions was center. noted in a month from ECP start. Disclosure of Interest: None declared. Conclusion: The efficiency of mononuclear cell collection is crucial for ECP immunomodulatory effects and results. AB045 However, optimal leucocytes isolation is very difficult to Comparison of prophylactic use of immunoglobulin obtain in patients with lipemic plasma. To our knowledge, M-enriched polyclonal immunoglobulin (Pentaglobins) this is the first reported case of TPE use to allow ECP treatment versus standard immunoglobulin after pediatric for GVHD in a patient with severe lipemia and very high levels hematopoietic stem cell transplatation of bilirubin. 1 2,* 1 1 3 1 References: 1) Cancer Immunol Immunother. 2014 F. Azik , C. Cayram , M. Erkoc¸og˘lu , H. Tezer , A. Erdem ,P.Is¸ık , Z. Avcı1,N.O¨ zbek2, B. Tavil1, B. Tunc¸1 Aug;63(8):757-77. Epub 2014 Jul 5. 1 2) Rev Bras Ter Intensiva. 2012 Sep;24(3):302-307. Ankara Children’s Hematology and Oncology Hospital, Ankara, Turkey, 2Ankara Children’s Hematology and Oncology Hospital, 3) Crit Care Nurse. 2013 Aug;33(4):18-23 3 4) G Ital Nefrol. 2012 Jan-Feb;29 Suppl 54:S31-5 Ankara, Trinidad and Tobago, Ankara Children’s Hematology Disclosure of Interest: None declared. and Oncology Hospital, Ankara, Togo Introduction: Patients undergoing allogeneic hemato- poietic stem cell transplant (HSCT) have a higher incidence of infections, possibly related to secondary hypo- Infectious complications gammaglobulinemia. Some centers use prophylactic intravenous polyclonal immunoglobulin (IVIG) to overcome this risk during the first 90 days after allo HSCT. There are few studies evaluating the use of IgM-enriched IVIG AB044 (Pentaglobins) in HSCT recipients in terms of prevention BK polyoma virus ascociated hemorrhagic cystitis and of infections, although its beneficial effects as adjunct nephritis in patients with undergoing allogeneic therapy in sepsis. Thus, we aimed to compare the efficacy hematopoietic stem cell transplantation of prophylactic use of IVIG versus prophylactic use of B. S. Karagun1,*, A. Eken2, B. Antmen1, I. Sasmaz1, M. Serbest1, Pentaglobins. B. Soyupak2, T. Akbas3, T. Arpaci3, N. Ozcan 3 Materials (or patients) and methods: We performed a 1Pediatric Hematology and Oncology, Adana Acibadem Hospital prospective, randomized study to compare the outcome Bone Marrow Transplantation Unit, 2urology, Adana Acibadem between use of prophylactic IVIG or Pentaglobins in patients Hospital urology, 3radiology, Adana Acibadem Hospital after allogeneic BMT. Eligible patients were those without radiology, Adana, Turkey documented infection at entry. This study excluded patients undergone bone marrow transplantation for primary immune Introduction: Hemorhagic cystitis (HC) is a common compli- deficiencies that might have hypogammaglobulinemia due to cation following allogeneic hematopoietic stem cell transplan- underlying disease. All transplants were related. Immunoglobu- tation (allo-HSCT). Several risk factors have been reported lin levels were measured pre-transplant, on day 0, 7,14,21,28 and including BU-containing myeloablative conditioning, unre- then monthly post BMT in 59 patients. Randomized patients are lated donors and GVHD. In contrast to early-onset HC, late classified as group 1 (IVIG) and group 2 (Pentaglobin) The first onset HC mainly results from polyoma BK virus (BKV) dose of IVIG or Pentaglobins was administered before and less frequently adenovirus or cytomegalovirus. conditioning regimen, on day þ 1, þ 8, þ 15, þ 22. And then, Reactivation of human polyomavirus BK (BKV) may cause was given if IgG level was below 400 mg/dl. All patients in both polyomavirus-associated nephropathy or polyoma virus-asso- groups were followed for the the first 100 days after allogeneic ciated hemorrhagic cystitis in bone marrow-transplant HSCT in terms of infection prevention. patients. Results: Twenty-seven patients in group 1 and 32 patients in We present 9 patients with BK polyoma virus (BKV) ascociated group 2 were included in the study. Patients in both groups hemorrhagic cystitis and 2 patients with BK polyoma virus were similar except for myeloid engraftment time, which was associated hemorrhagic cystitis and nephritis. longer in group 2. There were no siginificant differences in the Materials (or patients) and methods: Between 2013 and duration of neutropenia, hospitalization, fever, and in the 2014, 51 patients received an allogeneic BMT at Acıbadem number of pyrexial episode, septicemia and bacteriemia, CMV Adana Hospital Pediatric Bone Marrow Transplantation Unit. infection, acute graft-versus-host disease, veno-occlusive 11 patients occurred BKV associated hemorrhagic cystitis and disease, adverse events between the IVIG group and nephritis. BKV was detected in the urine analysis and blood by Pentaglobin group. PCR(polymerase chain reaction) in all patients. Conclusion: The benefit of prophylactic administration Results: We presented 11 patients with BKV infection, age of IVIG after allo HSCT remains controversial. Centers for ranging from 3 to 20 with a average of 12.8 years. They Disease Control recommend administering IVIG every 1 to 2 underwent allogeneic BMT due to thalassemia major (7 weeks in patients whose IgG level iso400 mg/dl to maintain a patients), aplastic anemia (3 patients) and acute lymphoblastic serum IgG concentration of4400 to 500 mg/dl (level of leukemia (one patient). The patients were treated with recommendation, BII). At present, based on a recent meta- hydration, continous bladder irrigation, ciprofloxacin, and analysis, routine administration of IVIG as prophylaxis within weekly intravesical hyaluronic acid instillation for four weeks, the first 100 days after allogeneic HSCT is not recommended, and cidofovir. Ten patient showed complete resolution of except high-risk allo HSCT recipients with severe hypogamma- hematuria. One patient with reft-ractory above these therapy globulinemia (IgG o400 mg/dl). In the current study, no also received hyperbaric oxgen and recombinant factor VIIa significant differences were noted between the utilization of (rFVIIa, NovoSeven; Novo Nordisk,Bagsvaerd, Denmark). Hemo- IVIG and Pentaglobin within the first 100 days after allogeneic dialysis was performed in two patients who developed renal HSCT. The major limitation of the current study was the lack of failure due to nephritis. a placebo control group, and thus we didn’t analyse if the Conclusion: BKV is ubiquitously present in the general utilization of IVIG or Pentaglobin has beneficial effect over population. Reactivation of infection occurs under placebo. conditions of immunosuppression; particularly HSCT or renal Disclosure of Interest: None declared.

S573 AB046 4. Kamble RT et al. Human metapneumovirus infection in a Human metapneumovirus in haematopoetic stem cell hematopoietic transplant recipient. Bone Marrow Transplant. transplantation (HSCT) recipients; a case series 2007; 40:699-700. E. De Jongh1,*, B. Hoppe2 Disclosure of Interest: None declared. 1VU Medical center, Amsterdam, 2Pulmunology, Leiden Medical center, Leiden, Netherlands AB047 Ochrobactrum anthropi bacteraemia after allogeneic stem Introduction: Human metapneumovirus (hMPV) is an envel- cell transplantation: Report of a newly emerging pathogen oped, single-stranded RNA virus, first detected in 2001 (1). It is F. Hindilerden1,*, T. Ozcelik2, S. Koculu3, H. S. Goksoy1, S. Guvenc1, a member of the Paramyxoviridae family, including respiratory R. Diz Kucukkaya1, M. Arat2 syncytial virus (RSV) and parainfluenza virus (PIV). hMPV is 1Istanbul Bilim University, Department of Internal Medicine distributed worldwide and thought to be transmitted by direct Division of Hematology, 2S¸is¸li Florence Nightingale Hospital or close contact with contaminated secretions, such as saliva Adult Hematopoietic Stem Cell Transplantation Unit, 3Istanbul or large particle aerosols (2), with an incubation period Bilim University Department of Infectious Disease and Micro- estimated to be 4 to 6 days (3). Symptoms range from mild biology, Istanbul, Turkey upper respiratory tract infection to severe pneumonia requir- ing mechanical ventilation. Patients with hematologic dis- Introduction: Ochronobactrum Antropi is an emerging eases, especially hematopoietic stem cell transplantation oppurtunistic pathogen in immuncompromised hosts, parti- (HSCT) recipients are likely to be at increased risk of infection cularly in patients with indwelding catheters as well as in with a prolonged clinical course (4). immunocompetant indiviuals. There had been rare reports of Materials (or patients) and methods: We describe our nosocomial bacteraemia caused by O.Antropi in solid organ experience with three HSCT patients diagnosed with hMPV transplant recipients (kidney, liver and pancreas). To our infection. These three patients were part of a much larger knowledge, O.Antropi bacteremia following allogeneic hema- group of patients diagnosed with hMPV infection in that same topoietic stem cell transplantation (HSCT) has not been time frame in our hospital (Table 1). previuosly described. Results: Materials (or patients) and methods: A 42- year-old man was Conclusion: hMPV is an important pathogen causing respira- admitted for fever and bloody diarhea 4 months after matched tory tract infections. Especially immunocompromised patients related allogeneic HSCT for refractory chronic lymphocytic are at risk of developing severe respiratory complications, with leukemia. At time of admission, he was under maintenance considerable morbidity and mortality. While considerable corticosteroid treatment for acute skin and liver GVHD. His progress has been made at the diagnostic level, proven temperature was 39rC, the blood pressure was 80/60 mmHg. treatment is still lacking. Ribavirin remains the only drug that Results: Blood was drawn for cultures and intravenous (IV) has been used in humans to treat hMPV infection, but in the meropenem and vancomycine were initiated. Unresponsive to absence of randomized studies it is impossible to conclude vigoruos IV fluid hydration, IV inotropic support was initiated with certainty on the efficacy of ribavirin. The development of in intensive care unit. Due to develeopment of hypoxemia and a is desirable and ongoing studies are promising. diffuse interstitial lung infiltrate of the left lung on X-ray, IV References: 1. van den Hoogen BG et al. An newly discovered TMP-SMX and gancylovir 5 mg/kg/day were started. Enta- human pneumovirus isolated from young children with moeba histolytica antigen in stool was positive and IV respiratory tract disease. Nat Med 2001; 7:719-724. metronidazole was added. Patient defervesced on 4th day of 2. Kim S et al. Molecular epidemiological investigation of a antibiotherapy and inotropic support was comletely halted. Of nosocomial outbreak of human metapneumovirus infection in the 3 sets of blood cultures obtained at 48-hr intervals, 2 a pediatric hemato-oncoloy patient population. J.Clin.Microbiol. bottles from Hickman-type central venous catheter lumen and 2009; 47:1221-1224. 1 bottle from peripheral vein yielded gram-negative rods. The 3. Peiris JS et al. Children with respiratory disease associated microorganism was identified as Ochronobactrum Antropi with metapneumovirus in Hong Kong. Emerg Infect Dis 2003; using the Vitek 2 automated system. Susceptibility testing 9: 628-633. showed the microorganism to be susceptible to carbapenems,

[AB046]

Age Hematologic Treatment ANC count at Pulmonary ICU Treatment Status Cause of death disease diagnosis characteristics Admission

57 Multiple autologous SCT 3,18x10(9) bilateral Yes No Deceased Death myeloma consolidations (3-3-2014) of disease 62 Acute induction 8,86x10(9) bilateral Yes ribaverin/ Deceased Relapse myeloid leukemia therapy 1st cycle consolidations IVIG (7-4-2014) of CVA 67 Plasma allogeneic SCT 0,42x10(9) bilateral Yes ribaverin/ Alive No cell leukemia consolidations IVIG 60 Acute induction 0,00x10(9) bilateral No No Alive No myeloid leukemia therapy 2nd cycle consolidations 62 Acute autologous SCT 0,00x10(9) pneumonia No No Alive No myeloid leukemia right lower lobe 70 Acute induction 0,00x10(9) bilateral Yes ribaverin/ Deceased Lung bleeding myeloid leukemia therapy 2nd cycle consolidations IVIG (02-04- 2014) 62 Acute lymfocytic consolidation 3,77x10(9) pneumonia No No Alive No leukemia therapy middle lobe 66 Acute induction 0,01x10(9) bilateral Yes ribaverin/ Deceased Herpes myeloid leukemia therapy 1st cycle consolidations IVIG (17-05- encephalitis 2014) 68 Multiple autologous SCT 0,00x10(9) pneumonia Yes No Deceased Multiorgan myeloma right lower lobe (13-03- failure, sepsis 2014)

S574 aminoglycosides and fluoroqionolones and resistant to and pneumonia was detected in 13.3% of all episodes. No B-lactams, TMP-SMZ and colimycine. Consecutive fecal speci- fungal or viral pneumonia was observed. Mean erythrocyte, mens obtained during and after bacteremia remained sterile. apheresis thrombocyte and random thrombocyte suspensions Meropenem was switched to ciprofloxacine after 2 weeks to that used during FNEs was 3.8, 3.5, 3.8 units respectively. complete the total duration of therapy to 3 weeks. Repeat Conclusion: It can supply useful additive for a better FENs blood cultures remained negative. Metronidazole was com- management process if the medical centers follow their pleted to 3 weeks. Colonoscopy performed on 2nd week of infection agents closely, perform CMV and SGA assays and admission for ongoing hematochesia identified a CMV DNA modify their empiric antibiotic treatment policies. load of 10703 copies/ml from biopsy samples. Gancylovir 2x5 Disclosure of Interest: None declared. mg/kg/day was begun on 12th day of admission and completed to 30 days. AB049 Conclusion: There is no consensus about the clinical features Frequency of HHV-6 and HHV-7 infections activation in and the best treatment strategy for O.antropi bacteremia. The patients with autologous and allogeneic peripheral blood most effective treatment modality has been remmoval of stem cell transplantation indwelding catheters. In rare cases of O.Antropi bacteremia 1,* after solid organ transplantation, contamination of conditioning I. Trociukas on behalf of 1Riga East Clinical University Hospital, regimen drugs during or after the manufacturing phase and Chemotherapy and Haematology Clinic, Riga, Latvia 2 A.Kirchen- stein Institute of Microbiology and Virology, Riga Stradins spread from gastrointesinal tract had been reported as potential 2 3 1 sources of contamination. Our patient presented with septic University, Riga, Latvia, S. Chapenko , M. Abolina , S. Lejniece , M. Murovska2 shock due to catheter-related bacteremia. A brisk response to 1 therapy was achieved without catheter removal despite the 1Riga East Clinical University Hospital, Chemotherapy and copresence of a resistant microorganism and immunsupression Haematology Clinic, Riga, Latvia 2 A.Kirchenstein Institute of triggered by CMV infection and GVHD in the setting of Microbiology and Virology, Riga Stradins University, Riga, Latvia, 22A.Kirchenstein Institute of Microbiology and Virology, Riga allogeneic HSCT. Further reports are needed to understand the 3 significance of this new emerging pathogen in allogeneic HSCT. Stradins University, Riga, Latvia, 1Riga East Clinical University References: 1. Ezzedine H, Mourad M, Van Ossel C, Logghe C, Hospital, Chemotherapy and Haematology Clinic, Riga, Latvia, Squifflet JP, Renault F, Wauters G, Gigi J, Wilmotte L, Haxhe JJ. Riga, Latvia An outbreak of Ochrobactrum anthropi bacteraemia in five organ transplant patients. J Hosp Infect. 1994 May;27(1):35-42. Introduction: HHV-6 and HHV-7 are ubiquitous (seropreva- Disclosure of Interest: None declared. lence rate 50-90%) T-lymphotropic immunomodulating viruses that after primary infection establish a life-time persistent infection and can be reactivate in immunosuppressed hosts, AB048 that may potentially cause immune dysregulation and Evaluation of febrile neutropenic episodes in autologous different post-transplant complications. Aim of this study peripheral blood stem cell transplantation for malignant was to evaluate 1) the incidence of persistent HHV-6 and HHV- solid tumors: experience of 30 cases 7 infection and frequency of these viruses infection activation G. Saydam1,*, M. Comert Ozkan1, F. Yilmaz1, N. Soyer1, in early period after auto- and related allo-PBSCT; 2) the A. P. Gorcegiz2, B. Arda3, R. Uslu2, F. Vural1 potential interaction with post-transplant complication devel- 1Hematology, 2Medical Oncology, 3Infectious Disease, Ege opment; 3) the potential interactions between both virusies. University,˙ Izmir, Turkey Materials (or patients) and methods: We analyzed the date of 56 patients: 44 patients (27 females, 17 males; mean age Introduction: In patients with advanced local or metastatic 34.1±10.7) underwent auto-PBSCT (group I) with HD - 29, NHL solid tumors, autologous hematopoietic stem cell transplanta- - 5, MM - 10 patients; 12 patients (6 females, 6 males; mean tion (HSCT) applied after high-dose chemotherapy is a rescue age 35.7±7.2) underwent related allo-PBSCT (group II) with therapy. The most important cause of mortality that mature AML-7, ALL– 3, HD – 2 patients. Viral genomic sequences in after HSCT is infections in neutropenic period, febrile PBL and cell-free plasma (marker of active infection) were neutropenic episodes (FNEs). The aim of this study was to detected by the nPCR, virus-specific antibodies - by ELISA, evaluate the incidence of FNEs and identify the infectious HHV-6 load – by quantitative real-time PCR, HHV-6 A and HHV- agents for determination of our treatment protocol. 6B - by restriction endonuclease analisis. Materials (or patients) and methods: Data of FENs in Results: Significant difference in the prevalence of anti-HHV-6 autologous HSCT for 30 patients with malignant solid tumor specific IgG class antibodies was not detected between auto- who was underwent transplantation at Hospital of Ege and allo-PBSCT patients before transplantation and grafts University, Adult Hematology Department, Transplant Center donors (35/44, 79.2% and 8/12, 66.7%, respectively). Simulta- between January 2009 and July 2014 were analyzed neous presence of IgM and IgG class antibodies was found in retrospectively. 2.3% (1/44) of I group patients and in 16.7% (2/12, P ¼ 0.11) of II Results: Twenty-nine of patients were male and one was group patients. Activation of single HHV-6 infection was not female. The mean age of the patients was 27.2 years (16-44 detected before and after PBSCT. The frequency of single HHV- years). The diagnosis of the patients were testicular cancer 7 infection activation after SCT was four time then before (germ cell; n ¼ 21, seminoma; n ¼ 3), ewing sarcom (n ¼ 3), transplantation (22.2% and 48.1%, respectively). The frequency medullablastom (n ¼ 2), and undifferentiated nasopharynx of concurrent (HHV-6 þ HHV-7) infection activation was sig- cancer (n ¼ 1). Central venous catheter was used in 24 nificantly higher after then before PBSCT (9.5% and 57.1%, (80.0%) of patients. FNEs was detected in 29 (96.6%) of HSCTs. respectively). The mean HHV-6 load before PBSCT was 6,781, All the patients received antiviral and antifungal prophylaxis whereas it was 54,676 copies/mg DNA during the period of and antibacterial prophylaxis was used in 20 (66.6%) of HSCTs. activation. Reactivation of HHV-7 preceded HHV-6 reactivation. In detected FNEs, 16.6% efficient pathogen microorganism The complications development (febrile neutropenia, gastro- was isolated from blood cultures. Sixty percent of isolated enteritis, pneumonia) was associated with the viruses reactiva- pathogens were gram-positive, 40% were gram-negative tion. Only HHV-6B was detected in all DNA positive samples. bacteria’s. No efficient pathogen microorganism was isolated Conclusion: The pathogenic role of HHV-7 still remain unclear from urine cultures. Twenty-five percent efficient pathogen as well as interaction between HHV-7 and HHV-6 by microorganisms were isolated from catheter cultures. Efficient concurrent infection. pathogens were isolated in 20% of stem cell products. Serum High frequency of HHV-7 and concurrent (HHV-6 þ HHV-7) galactomannan antigen (SGA) was detected positive in 6 (20%) infection activation, serum/plasma expression levels suggest of HSCTs that all of them were false positive. CMV-DNA was that both viruses are involved in the complications develop- negative in all episodes. Eleven HRCT was performed in FNEs ment in the early period after auto- and allo-PBSCT. The

S575 kinetics of the viruses reactivation reflects the potential role of characteristics in patients following autologous peripheral HHV-7 as co-factor of HHV-6 reactivation. Our data suggest blood stem cell transplantation (auto PBSCT). Between May that valacyclovir used in experience for antiviral prophylaxis 2009 and December 2013, 115 patients were underwent auto and therapy is not sufficiently effective to prevent of HHV-6 PBSCT in our Haematology Department (56 male, and 59 and HHV-7 infection activation after PBSC. female with median 54 year of age at transplantation). Majority Disclosure of Interest: None declared. of patients (n ¼ 68) were diagnosed with multiple myeloma (MM), and 13/35 with Hodgkin lymphoma (HL)/ non-Hodgkin AB050 lymphoma (NHL). The conditioning regimens for NHL and HL patients were according to BEAM protocol, melphalan 200 mg/ Stimulation of protein-reactive effector cells by 2 T-activated proteins: a novel strategy for monitoring cell m for MM cases. CMV seropositivity was observed in 52 cases mediated immunity (42%) pretransplant. All patients received antiviral prophylaxis 1 1 1 1 2,* (acyclovir 200 mg orally four times per day) for 3 months. S. Barabas , T. Spindler , T. Widmann , C. Tonar ,K.Bo¨ckl , During transplantation procedure and post-transplant 100- L. Deml1 1 2 day period, 876 plasma samples were collected from 115 Research & Development, Marketing&Sales, Lophius Biosciences patients to detect CMV DNA viral load. GmbH, Regensburg, Germany Results: Out of 876 samples 61 (6.9%) were positive from 41 patients (with median 1540 copies/ml). Six of Introduction: Suppression of cell mediated immunity them were treated with gancyclovir. The most common (CMI) may cause serious clinical complications including manifestations of CMV reactivation were fever, pancytopenia, microbial infections and reactivations. Thus, accurate quanti- prolonged platelet and neutrophil engraftments, and diar- fication and monitoring of functional effectors of CMI may rhoea. Only in one case, severe pneumonia was observed with allow reliable prediction, detection and personalized treat- fatal outcome. ment strategies. Conclusion: The use of quantitative CMV PCR assays are Materials (or patients) and methods: We have developed a essential for earlier recognition of CMV reactivation, to restrict novel methodology for the functional analysis of CMI based on the use of gancyclovir, to prevent selection of gancyclovir the combined application of acitvated herpesvirus-derived resistant strain, and to reduce the toxicity of this drug. stimulator antigens and the highly sensitive ELISpot Disclosure of Interest: None declared. technology. Results: Activated proteins reveal the unique capacity to stimulate a broad spectrum of clinically relevant effector cells AB052 including T helper and cytotoxic T cells as well as NK and NKT Granulocyte Transfusion in life threatening Infections in cells. The activation of proteins mediates the translocation to patients under hematopoietic stem cell transplantation the exogenous and endogenous processing pathway of APC M. Mehdizadeh1,* for epitope-loading on MHC-I in addition to MHC-II molecules 1Taleghani BMT center, Shahid Beheshti university of medical by cross-presentation. Stimulation of peripheral blood mono- sciences, Tehran, Iran, Islamic Republic Of nuclear cells (PBMC) with preselected activated CMV-proteins reveal a clinical sensitivity above 90% independent of the Introduction: life-threatening infections are not unusual in subject´s HLA composition. Assay performance is highly severe neutropenic patients, and the severity and duration of reproducible with variability values less than 25% and a linear neutropenia are significantly related to the mortality of correlation between the amount of reactive cells and total infections. PBMC counts. Granulocyte transfusion is an old option for patient with Conclusion: Thus, activated stimulator antigens in combina- severe infection and neutropenia that has been substituted by tion with ELISpot technology represent a promising strategy to novel antibiotics and granulocyte colony stimulating factor assess the immune status of immunosuppressed patients. although some patients have dismal situation that don’t Disclosure of Interest: S. Barabas Employee of: Lophius respond to these treatments especially patient severe aplastic Biosciences, T. Spindler Employee of: Lophius Biosciences, T. anemia or under hematopoietic stem cell transplantation. In Widmann Employee of: Lophius Biosciences, C. Tonar our experience, we set up the apheresis technique of Employee of: Lophius Biosciences, K. Bo¨ ckl Employee of: granulocytes and then we followed the efficacy and safety Lophius Biosciences, L. Deml: None declared. of granulocyte transfusion in two patients with resistant psodomonas infection and severe neutropenia who were under bone marrow transplantation. AB051 Materials (or patients) and methods: Granulocyte apheresis Monitoring Cytomegalovirus infection by quantitative technique and Transfusion real-time PCR method following autologous peripheral Granulocyte concentrates were ABO compatible to recipients stem cell transplantation and were collected with blood cell separator (Cobe K. Piukovics1,*, G. Terhes2, T. Gurbity Pa´lfi1, I. Marton1, spectra, US) from health donors after 3 days of granulocyte A´. Bereczki1, Z. Borbe´nyi1, J. Dea´k2, E. Urba´n2 Mobilization using GCSF 5 micro gram /Kg in Taleghani BMT 12nd Department of Internal Medicine and Cardiology Center, center.The mean granulocyte dose in concentrates was 2Institute of Clinical Microbiology, University of Szeged, Szeged, 9.264.76109 cells. Granulocyte concentrates were transfused Hungary within 4–6 h aftercollection. Patients received anti-anaphylaxis drugs before transfusions. Adverse effects of granulocyte Introduction: Human cytomegalovirus (CMV) infection is a transfusions included mild to moderate fever, chills, allergy major cause of morbidity and mortality among patients and dyspnea. underwent stem cell transplantation procedure. This may Results: we set up the apheresis technique of granulocytes arise as a primary infection or due to reactivation. It can occur and then we followed the efficacy and safety of granulocyte with lower incidence following autologous stem cell transfusion in two patients with resistant psodomonas transplantation, than in allogeneic setting. Following this infection and severe neutropenia who were under bone procedure, CMV serologic assay is important to determine marrow transplantation.In our patients there was no severe patient’s risk for CMV infection, but not for diagnosis of adverse event associated with granulocyte transfusions.We disease. Real-time quantitative PCR is primary and most observed that granulocyte transfusions in severe infections reliable method for surveillance of active CMV infection in with neutropenia is life saving in Two patients infected by this patient group. resistant pseudomonas who were cured successfully with Materials (or patients) and methods: Our aims were to granulocyte transfusions with GCSF,and their hematopoiesis evaluate occurrence of CMV infection, and epidemiological recovered later.

S576 Conclusion: Granulocyte transfusion might be an adjunctive fungi to high risk of IFD patients and use them individually in therapy for treating severe infections in neutropenic patients. moderate risk patients. The primary objective was the IFD free Disclosure of Interest: None declared. survival during the early period (100 days post-transplant) at a single center in allo-HSCT recipients not considered as high AB053 risk for IFD, during primary prophylaxis with fluconazole with a Hepatitis B virus (HBV) reactivation after allogeneic close clinical and microbiological vigilance. haematopoietic stem cell transplantation in a single center Materials (or patients) and methods: We have conducted a 1,* 2 1 2 retrospective descriptive study in 20 consecutive allo-HSCT M. Liga , C. Koutougeras , A. Karela , A. Spyropoulou , recipients not considered as high risk for IFD1, from a total of M. Tiniakou1, E. Triantafyllou1, A. Spyridonidis1 1 2 39 allo-HSCT from January 2013 to January 2014, in a single Bone Marrow Transplantation Unit, Microbiology, University institution. Patients were excluded because of history of Hospital of Patras, Patras, Greece possible or probable IFD (7), alternative or experimental transplants sources (6), or prolonged neutropenia prior to Introduction: Reactivation of HBV is a serious complication of conditioning regimen (6). HLA mismatch by itself was not chemotherapy/immunosuppressive therapy in patients with considered as high risk. During the neutropenia period, HBV infection. In this article, we aim to investigate the patients stayed in HEPA-filtered rooms, with assessment of frequency of HBV reactivation after allogeneic haematopoietic serum Aspergillus galactomannan antigenemia (AGA) twice a stem cell transplantation (HSCT) in a single center. week until neutrophil engraftment41 x 109 / L, and once a Materials (or patients) and methods: 111 patients were week thereafter. When persistent fever or positive AGA, chest transplanted in the University Hospital of Patras from October CT were performed, as well as other locations if symptoms. 2005 to July 2014, for haematological malignancies. Sixteen The IFD were classified according to the EORTC/MSG criteria. patients had resolved HBV infection ( ¼ negative HbsAg, Results: There were 16/20 (80%) episodes of febrile neutro- positive antiHBs and antiHBc antibodies), 38 patients were penia, but only in one patient lasted more than five days (5%). HBV naı¨ve ( ¼ negative HbsAg, negative antiHBs and antiHBc CT was performed in 4/20 (20%) patients. None of them antibodies), 47 patients were vaccinated ( ¼ negative HbsAg, presented signs of IFD. During the neutropenia period, we positive antiHBs and negative antiHBc antibodies), 2 patients identified positive serum AGA in 2/20 (10%) patients, one of were seropositive ( ¼ positive HBsAg) and in 8 patients data them studied by bronchoalveolar lavage (BAL) AGA, being were not available. Median age at transplantation was 40 years negative. We found no possible/probable/proven IFD. Regard- (range, 17–70), with 66 male and 45 female patients. Median ing to antifungal treatment, 3/20 (15%) patients initiated follow up of the cohort was 403 days (range, 0–2556). broad-spectrum antifungal pre-emptive therapy with Ampho- Results: Overall, 4 (3.6%) patients presented HBV related tericin B liposomal (AmB-L) (2 patients due to positivity of events. 3/4 had a history of resolved HBV infection and serum AGA) or empirical therapy with AmB-L (1 patient due to presented with HBV reactivation after HSCT on days þ 575, prolonged febrile neutropenia). Among patients who þ 294 and þ 761 respectively. All of them presented remained with fluconazole 7/17 (41%) received prophylactic with a clinical picture of acute HBV infection (positive HBsAg, extended spectrum azoles as primary prophylaxis due to high positive HBV-DNA, increased SGOT and SGPT) and none of dose steroid treatment for GVHD (6 patients) and broad them has received prophylaxis with lamivudine for HBV spectrum antifungal empirical therapy due to inespecific reactivation after the HSCT. All three cases were completely neurological symptoms (1 patient). None of them presented resolved after treatment with entecavir. The third patient signs of IFD in the 100 days post-HSCT follow-up. The use of was a seropositive one that received a graft from a fluconazole was found to be safety. We did not find drug seropositive sibling donor. The patient was HbsAg positive interactions or toxicities associated with its use that let to and HBV-DNA negative at transplant and received treatment treatment discontinuations. with entecavir until day þ 540 post HSCT, that was interrupted Conclusion: Fluconazole and the use of HEPA-filtered rooms from the patient’s own will. At that time he was still HBV-DNA should be considered as an effective antifungal prophylaxis negative. After 197 days of interruption of entecavir treatment strategy in patients undergoing allo-HSCT not considered as the patient presented with acute HBV infection (increased 8 high risk of IFD, with a good safety profile. Budget impact HBV-DNA 1.7 10 copies, increased total bilirubin, SGOT, analysis regarding the use of extended-spectrum antifungal SGPT, ALP and g-GT). Treatment with entecavir was recom- prophylaxis in this subgroup of patients is being made at this menced but because of slow resolution of the acute HBV time. infection, tenofovir was added to his treatment. The patient References: 1. Barbera´nJ,et al. Rev Esp Quimioter. 2011; currently is receiving the above treatment for three months, 24:263-70 with an improvement of 3 log of the HBV-DNA, which is still Disclosure of Interest: None declared. positive. Conclusion: HBV reactivation remains a problem after allogeneic HSCT. Prophylaxis is recommended, in patient with AB055 resolved HBV infection and in seropositive patients although Evaluation of Dynamic HBV-DNA Levels in patients the optimal duration of prophylaxis remains to be determined. undergoing hematopoietic stem cell transplantation with Disclosure of Interest: None declared. high risk for HBV infection E. Abdullayev1, C. Hasanzade1, S. Berksoy1,*, G. Birol1, M. Ko¨ksal Erkis¸i1 AB054 1Adult Bone Marrow Transplantation Unit, Cukurova University Primary Prophylaxis with non-extended spectrum azoles Faculty of Medicine, ADANA, Turkey in Allogeneic Hematopoietic Stem Cell Transplant Recipients Introduction: Background: N. Revilla1,*, V. Garcı´a-Gutie´rrez1, P. Herrera1, J. A. Garcia-Vela1, It is mentioned that HBV-DNA PCR titration may show the J. Fonseca1, I. Saavedra1, A. Jime´nez1,J.Lo´pez-Jime´nez1 patients with high risk for HBV infection prior hematopoietic 1Hematology, Hospital Ramon y Cajal, Madrid, Spain stem cell transplantation (HSCT). Aim: We planned a study for analyising our HBsAg( þ ) and/or Introduction: Invasive fungal disease (IFD) remains a major Anti-HBc( þ ) patients whether they have risk for HBV infection cause of morbidity and mortality in allogeneic hematopoietic by measuring dynamic DNA levels. stem cell transplant recipients. Antifungal prophylaxis during Materials (or patients) and methods: Patients who were the early period of the allogeneic hematopoietic stem cell candidates for autologous or allogeneic HSCT were conducted transplantation (allo-HSCT) should be individualized according to the study. HBV serological markers were investigated in all to risk groups, leaving antifungal agents against filamentous patients. Patients with HBsAg( þ ) or Anti-HBc( þ )/Anti-HBs(-)

S577 were included to study. HB-DNA levels were measured with acute phase reactant that is used as a marker of PCR method in at least 3 times, prior conditioning, post associated bacterial infection. The aim of this study was to conditioning and post stem cell infusion. evaluate the usefulness of CRP as a diagnostic marker and Results: We conducted 38 patients. Two of the 38 patients prognostic factor for the febrile episode in haematological seemed to have HBV infection risk. One had HBsAg positivity patients. and the other had Anti-HBc positivity but both of them had Materials (or patients) and methods: Patients with febrile high DNA PCR titrations and were receiving anti-viral treatments episode managed in the haematology department were including lamivudine and tenovofir. The patients characteristics consecutively enrolled in the year 2013. Clinical characteristics were shown in Table 1. The results were shown in Table 2. and laboratory features including antibiotic prophylaxis, duration of fever, antibiotic treatment, and clinically or Table. 1 microbiologically documented infection were retrospectively analysed. The CRP was evaluated at the day of the onset of Patient Diagnosis Age/Gender HBsAg Anti-HBs Anti-HBc HBV status fever, the day of maximum CRP and the temporal difference between both. The differences between CRP in transplant 1. NL 32/M 3152 2 0,12 Chronic patients and non-transplant patients were analysed compar- 2. NHL 57/M 0,36 2 0,006 Acute ing mortality, microbiologically documented infection and hemodynamic stability. Conclusion: There has been limited data about the correlation Results: During the year 2013, 400 patients were admitted at between HBV infection risk and the level of HBV-DNA titration the hematologic ward. All febrile episodes (n ¼ 200) occurring in patients who are candidates for HSCT. in 27 transplant patients and 82 non-transplant patients were Although we have very few patients as a new transplantation investigated. The medium age of the study population was center in Turkey, we consider sharing our early experience 60,2 years (range 18-92). 48% were women. 12,5% underwent an autologous stem cell transplantation (n ¼ 25) and 10,5% received an allogenic stem cell transplantation (n ¼ 21). In all cases the source of hematopoietic progenitors was peripheral blood. The medium value of the result of CRP measurement performed on the onset of fever was 9,5 mg/dl (median: 6,65 mg/dl) the maximum medium value of CRP was 16 mg/dl (median 16 mg/dl). The medium value for the temporal difference between both was 1,86 days. Death patients had a significant higher maximum CRP than alive patients (21 mg/dl versus 15 mg/dl, P ¼ 0.0052). Every rise of a unit of CRP increased the mortality a 6%, being higher in transplant patients (12%). Hemodynamically with dynamic HBV-DNA titration. We think that not only HBV unstable patients had a CRP value significantly higher DNA levels but also antiviral treatment regimens, and than hemodynamically stable patients (22,7 mg/dl versus chronicity of HBV infection are important factors for HBV 15 mg/dl, Po0.0001). This signification remained in transplant infection risk in patients undergoing HSCT. patients. References: 1.Hande H Tuncer, Naveed Rana, Cannon Milani, All the patients with microbiological documented infection Angela Darko, and Samer A Al-Homsi et.al World J showed a CRP significantly higher than those patients without Gastroenterol. Apr 28, 2012; 18(16): 1851–1860. microbiological documented infection (21 mg/dl versus 2.Mikulska, M.; Nicolini, L.; Signori, A.; Rivoli, G.; del Bono, V.; 15 mg/dl, P ¼ 0.0013). This signification remained in the Raiola, A.M.; di Grazia, C.;Dominietto, A.; Varaldo, R.; Ghiso, A.; transplant patients as well. et al. Hepatitis B reactivation in HBsAg-negative/HBcAb- Conclusion: This study confirms the association between positive allogeneic haematopoietic stem cell transplant persistence of an elevated CRP value and unfavourable recipients: Risk factors and outcome. Clin. Microbiol. Infect. outcome of any febrile infectious episode in haemato- 2014, doi:10.1111/1469-0691.12611. logical patients since the rise of CRP was related to the 3.Liang R. How I treat and monitor viral hepatitis B infection in development of the febrile episode. CRP monitoring patients receiving intensive immunosuppressive therapies or could help us to identify those patients with bacterial undergoing hematopoietic stem cell transplantation. Blood. infections therefore start promptly the antibiotic treatment 2009;113:3147–3153. [PubMed] to improve the clinical course of the infectious episode and 4.Shingo Nakamoto, Tatsuo Kanda, Chiaki Nakaseko, Emiko the patient’s outcome. Sakaida, Chikako Ohwada,Masahiro Takeuchi, Yusuke Takeda, Disclosure of Interest: None declared. Naoya Mimura, Tohru Iseki, Shuang Wu,Makoto Arai, Fumio Imazeki, Kengo Saito, Hiroshi Shirasawa, Osamu Yokosuka et. al AB057 Reactivation of Hepatitis B Virus in Hematopoietic Stem Cell Bone marrow transplant for low risk thalassemia patients Transplant Recipients in Japan: Efficacy of Nucleos(t)ide in low resource setting Analogues for Prevention and Treatmet. Int. J. Mol. Sci. 2014, 1,* 15, 21455-21467; doi:10.3390/ijms151121455 S. I. Fatima on behalf of Bone Marrow Transplant for Low Risk Disclosure of Interest: None declared. Thalassemia Pa-tients in Low Resource Setting ‘‘The Children’s Hospital Pakistan Institute of Medical Science, Islamabad Naila Yaqub,Syeda itrat Fatima,Sadaf Khalid,Tatheer Zahra, Sara AB056 Gilani, Abstrac Usefulness of C-reactive protein as a diagnostic and 1cure2children foundation Pakistan, Children hospital( Pakistan prognostic marker in haematological patients with febrile institute of medical sciences Islamabd), Islamabad, Pakistan episode S. Monsalvo1,*, V. Teresa1, L.-L. Jose-Luis1, R.-G. M.Victoria1, Introduction: To evaluate performance of a newly developed P.-S. M.Angeles1, A. Elham1, A. Teresa1, Y. Maria1, M. Raquel1, low resource Bone Marrow Transplant unit for Thalassemia T. Claudia1, P. Llamas1 major in Islamabad Pakistan. 1Fundacion Jimenez Diaz, Madrid, Spain Materials (or patients) and methods: SUMMARY: Bone marrow transplantation (BMT) is the definitive cure with Introduction: Infection is an important cause of mortality in improved health and better out come of life in over 90% of the haematological patient. C-reactive protein (CRP) is an low-risk children with a matched related donor. This study

S578 retrospectively describes BMT complications in low risk This comprised cases of possible, probable and proven IFI patients (defined as livero2.5cm and ageo7y) with severe at rates of 4%, 3% and 2%, respectively. Univariate analysis thalassemia (ST) (defined as a thalassemia syndrome with showed that ATG (P ¼ 0.024) and Fludarabine usage spontaneous hemoglobino7 g/dL). (P ¼ 0.005) were possible risk factors. Fludarabine usage Most common complication faced during transplant was (relative risk factor for IFI ¼ 5.208, 95% confidence interval Infection. 1.346-20.000) is the only factor associated with IFI in Materials and Method; multivariate analysis. At the end of six months after Between januart 2009 and October 2014, low risk forty one transplantation, 25 of 119 (21%) children died after a median patients were transplanted. Retrospectively,medical Records of of 91 days (range, 14-180 days). Overall mortality rate bone marrow transplant of forty one low risk thalasemia major attributable to IFI at six months after allogeneic HSCT patients were evaluated for complication. was 2% (2/119) and IFI was responsible or co-responsible Results: Results: Thirty five had successful engraftment with for 8% (2/25) of all deaths. transfusion free survival accounting for 85.36%. Thirty eight Conclusion: In pediatric patients with acute leukemia under- patients had overall survival which is 92.68% Three patients going allogeneic HSCT, one should be aware of IFI particularly which is 7.31% had transplant re-lated mortality whereas same during the first six months when Fludarabine was used as part number of patients which is again 7.31% had failure of of conditioning regimen. engraftment. The manageable complications included hyper- Disclosure of Interest: None declared. tension and hemorrhagic cystit. 1 patient had veno occlusive disease.Most common complication was febrile neutropenia and culture proven infection ( either of blood or urine or of central venous catheter site). Conclusion: Conclusion: Bone Marrow transplant for transfu- Early complications / late effects sion dependent Thalassemia is possible in low resource setup & quality of life with results that are comparable with the centers in developed countries and the complications are manageable. The cost of transplant is 10,000 to 15,000 $ which is equivalent to three years of supportive treatment. AB059 Key word: Bone Marrow transplant, Thalassemia major, Low Safety of Hematopoietic cell transplant comorbidity resource Setup. index in prediction of survival after autologous Disclosure of Interest: None declared. transplantation in multiple myeloma – single center experience A. Pivkova Veljanovska1,*, S. Genadieva Stavric2, Z. Stojanoski1, AB058 L. Chadievski1, L. Cevreska1, B. Georgievski1 Evaluation of risk factors and outcomes of invasive fungal 1University Clinic of Hematology, BMT Unit, 2University Clinic or infections in the first six months after allogeneic Hematology, BMT Unit, Skopje, Macedonia, The Former Yugoslav hematopoietic stem cell transplantation in pediatric acute Republic Of leukemia: A Turkish multicenter study V. Hazar1,* on behalf of The Turkish Pediatric Bone Marrow Introduction: Both autologous stem cell transplantation Transplantation Study Group, G. Karasu2, G. Ozturk3, (ASCT) and novel agents have improved outcomes for patients S. Caki Kılıc2, V. Uygun4, M. Karakukcu5, N. Ozbek6, O. Gursel7, (pts) with multiple myeloma (MM). ASCT is often restricted to D. Atay3, A. Kupesiz8, S. Aksoylar9, E. Ozyurek10, U. Kocak11, fitter pts due to concerns of excessive treatment-related H. Daloglu4, E. Unal12, S. Yilmaz13, E. Unal5, I. Eker7, B. Tunc6, morbidity and nonrelapse mortality (NRM). Pre-transplant risk- F. Erbey3, N. Eker8, S. Kansoy9, T. Fiskin10, Z. Kaya11, H. Oren13, stratification based on comorbidity index (CI) has been G. Sezgin14, M. Ertem12, E. Kurekci7, A. Tanyeli14, E. Unal12, recognized as an important decision-making tool in pts with A. Yesilipek2 MM. Hematopoietic cell transplant comobrbidity index (HCT- 1Pediatric BMT Unit, Medipol University Faculty of Medicine, CI) was shown to predict survival and NRM in allogeneic 2Pediatric BMT Unit, Bahcesehir University Faculty of Medicine- transplantation. The role of HCT-CI in prediction of outcome Medical Park Gostepe Hospital, 3Pediatric BMT Unit, Medical Park during autologous setting for MM pts has to be more Bahcelievler Hospital, Istanbul, 4Pediatric BMT Unit, Bahcesehir evaluated and is still unclear. University Faculty of MedicineMedical Park Antalya Hospitali, Materials (or patients) and methods: We analyzed outcomes Antalya, 5Pediatric BMT Unit, Erciyes University Faculty of of 74 patients with MM treated at our hospital with ASCT after Medicine, Kayseri, 6Pediatric BMT Unit, Ankara Child Health and high dose melphalan during 12 years of center experience. Diseases Hematology and Oncology Training and Research The impact of HCT-CI, Karnofsky performance scores (KPS), Hospital, 7Pediatric BMT Unit, Gulhane Military Medical Academy, ECOG and other prognostic factors (cell dose, time to Ankara, 8Pediatric BMT Unit, Akdeniz University Faculty of transplant) on NRM and survival were studied in multivariate Medicine, Antalya, 9Pediatric BMT Unit, Ege University Faculty Cox regression models. of Medicine,˙ Izmir, 10Pediatric BMT Unit, Medical Park Samsun Results: Median age of patients was 51 yrs (40-65) with Hospital, Samsun, 11Pediatric BMT Unit, Gazi University Faculty of median time from diagnosis to ASCT 13 months (4-46). 59% of Medicine, 12Pediatric BMT Unit, Ankara University Faculty of pts were in first complete or partial remission prior to ASCT Medicine, Ankara, 13Pediatric BMT Unit, Dokuz Eylul University and 66% of pts had received 41 line of treatment prior ASCT. Faculty of Medicine,˙ Izmir, 14Pediatric BMT Unit, Cukurova All patients received melphalane-based preparative regimen. University Faculty of Medicine, Adana, Turkey HCT-CI of 0 had 40% of pts, HCT-CI scores 1-2 had 31% of patiens and HCT-CI score 43 had 29% of pts with MM. HCT-CI Introduction: Although invasive fungal infections (IFI) are a low risk were 40% of pts, median risk HCT-CI was noted in 31% major cause of morbidity and mortality after allogeneic of pts and high risk HCT-CI in 29% of pts. Higher HCT-CI was hematopoietic stem cell transplantation (HSCT), data for associated with lower KPS o90 (30% pts score of 0 versus 50% pediatric patients is scarce. in HCT-CI score 42). Factors that affect overall survival (OS) in Materials (or patients) and methods: We performed a MM pts were months from diagnosis to ASCT (Po0.001), retrospective review of 119 children with acute leukemia median and high HCT-CI (P ¼ 0.03), KPS (P ¼ 0.04), CD34 cells undergoing allogeneic HSCT to determine the incidence and o3,0x10(6)/kg (P ¼ 0.009). In multivariate analysis OS was risk factors for IFI and outcomes in the period between 1st of inferior in pts with HCT-CI score of 1 to 2 (RR 1.36, 95%CI, 1.01 July 2013 and 31st of December 2013. to 1,86 and P ¼ 0,04) and HCT-CI42 (RR 1.5, 95%CI, 1.09 to Results: A total of 11 episodes of IFI were observed in 119 2.05 and P ¼ 0.01). Factors that affect one year NRM in MM pts subjects (9%) in the first six months after allogeneic HSCT. were HCT-CI (P ¼ 0.03), KPS (P ¼ 0.02), hospital stay (P ¼ 0.02)

S579 and ECOG (P ¼ 0.0009). High and median HCT CI were Although our patient number is limited and follow-up period associated with NRM of 6% (P ¼ 0.001), respectively low HCT- very short, our preliminary results indicate that development CI pts had NRM of 2%. of cGvHD and associated GVT effect, may be an important Conclusion: ASCT for MM is associated with low NRM and factor in improving OS. death is related due to disease progression. Our data confirms Disclosure of Interest: None declared. the safety of ASCT in MM pts who have high HCT-CI due to low NRM. AB061 Disclosure of Interest: None declared. Defibrotide for the prevention and treatment of hepatic veno-occlusive disease after hematopoietic stem cell AB060 transplantation; A single center experience Preliminary results of acute lymphoblastic leukemia B. A. Antmen1,*, I. Sasmaz1, B. Karagun1, M. Serbest1, G. Ucar1, patients undergoing allogeneic hematopoietic cell T. Baysal1 transplantation: A single center experience 1Pediatric Hematolology, Adana Acibadem Hospital Bone A. I. E. Tekgu¨ndu¨z1,*, O. Kayıkcı1, S. Civriz Bozdag2, A. H. Kaya1, Marrow Transplantation Unit, Adana, Turkey S. Kocubaba1, S. Namdaroglu1, B. Ugur1, F. Altuntas1 1Hematology and bone marrow transplantation unit, Introduction: Hepatic veno-occlusive disease (VOD) is a Ankara Oncology Hospital, 2Hematology and bone marrow common and serious complication of hemotopoietic stem cell transplantation unit, Ankara University Hospital, Ankara, Turkey transplantation (HSCT) in children. We aimed to assess prospectively the use of prophylactic defibrotide in pediatric Introduction: Compared to pediatric age group the prognosis patients undergoing HSCT. of acute lymphoblastic leukemia (ALL) in adults is still dismal. Materials (or patients) and methods: Allogeneic hematopoietic cell transplantation (allo-HCT) is an In this study, 43 patients who underwent HSCT were given established treatment modality with curative potential in defibrotide prophylaxis as 25 mg/kg per day in four divided upfront and relapsed/refractory settings of adult patients with intravenous infusions over 2 h, starting on the same day as the ALL. Here we report our experience in adult ALL patients who pretransplantation conditioning regimen. The mean duration undergone allo-HCT. of use of defibrotide is 20 days as a prophylaxis. Materials (or patients) and methods: All consecutive Results: In this study, 43 patients were recruited, 27 male patients who were diagnosed with ALL and received allo- patients and 16 female patients, with the average of 8.8 years, HCT between January 2009 and December 2013 were range 1-20; 9% infants, 49% children and 42% adolescent. included in the study. Medical records of patients were There were 24 patients with thalassemia major, 11 patients retrospectively analyzed.Descriptive statistics are presented as with leukemia, 5 patients with aplastic anemia, one patient median and range. Survival analysis was calculated with with Diamond Blackfan anemia, one patient with congenitale Kaplan Meier analysis. A p value below 0.05 was considered to dyserythropoetic anemia and one patient with Kostman be statistically significant. syndrome. All transplants were allogeneic. No serious side Results: A total of 51 patients (female (n: 20); male (n:31)) effects were seen. In seven patients developed clinical VOD participated in the study. Media age of study cohort was 26 (Seattle criteria). In these patients, defibrotide dose was (17-58). The primary diagnoses were B-ALL and T-ALL, in 29 increased to a treatment dose of 40-60 mg/kg per day. One (54%) and 25 (46%) patients, respectively.Thirteen (25%) infant patient with Kostman syndrome died due to hepatic patients were Ph þ ALL.Stem cell source was peripheral blood and pulmonary veno-occlusive disease. After 24 months of in 44 (86%) patients, while 7 (14%) patients received bone follow up, 6 patients who developed VOD are being well and marrow derived hematopoietic cells.Forty-four (86%) and no patient have transplant related complications. seven (14%) patients received myeloablative and reduced- Conclusion: In this prospective study, we demonstrated that intensity conditioning regimens, respectively. Most of the the use of defibrotide is safe and effective in preventing and patients (n:44; 86%) were treated with allo-HCT using 6/6 HLA treating VOD in pediatric patients at high risk. matched-sibling donors. On the other hand 1, 1 and 5 patients Disclosure of Interest: None declared. received HCT from 5/6 HLA matched-sibling, 10/10 HLA matched-unrelated and haploidentical donors, respectively. AB062 Median EBMT score of patients was 2 (0-5). According to HCT- Abstract Withdrawn comorbidity index (HCT-CI), 25, 19 and 7 patients belonged to low (score:0), intermediate (score 1-2) and high risk (scoreZ3) groups, respectively. At HCT 5 (10%) patients were in CR2 and 14 (27%) patients received HCT in active disease. Median AB063 follow-up of study cohort was 14 months (1-58). During study Progression of heart AL amyloidosis after stem cell period 17 (33%) patients developed aGvHD. Chronic GvHD mobilization with filgrastim and plerixafor were observed in 11 (22%) patients. Transplant related D. Pohlreich1,*, J. Krejci2, J. Straub1, B. Vackova1, I. Spicka1, mortality at D þ 100 and 1 year were 5% and 11%, P. Nemec3, M. Trneny1 respectively. One year overall survival of study cohort was 1GENERAL TEACHING HOSPITAL, CHARLES UNIVERSITY, I. DEPT. found to be 62.7±7.On univariate analysis, we observed that OF HEMATOONCOLOGY, Prague, 2Department of Cardiovascular the development of cGvHD significantly improved one year OS Diseases, St. Anne´s University Hospital - ICRC, Masaryk University, (53±8vs90±8; p:0.039), while EBMT risk score, HCI-CI, 3Centre for Cardiovascular and Transplant Surgery, Brno, Czech development of aGvHD and Ph þ disease had no statistically Republic significant impact on OS. But on multivariate analysis, cGvHD lost its influence on OS. Introduction: 64-years old healthy patient, started complain- Conclusion: As the prognosis of relapsed/refractory ALL is ing about 4 months developing fatigue and shortness of extremely poor, all attempts should be made for providing the breath, coming to the hospital with dyspnoea according NYHA most effective antileukemia consolidation upfront. Therefore, II-III classification. Transthoracic echocardiography (TTE) some guidelines suggest allo-HSC for all eligible patients revealed hypertrophic cardiomypathy with normal left ven- following CR1. The most important factor decreasing relapse tricle function (EF 60%), mild diastolic dysfunction and risk in allo-HCT is its biologic effect on leukemia, which is suspicion of amyloidosis. Performed magnetic resonance of called graft versus leukemia (GVL) effect. It is widely accepted heart was in agreement with TTE. Lab results showed BMP 341 that sensitivity of ALL to GVT effect is moderate at best. If we ng/l, Troponin I 0,37 ug/l, monoclonal gamapathy IgG lambda consider that 37% of our patients were beyond CR1 at HCT, with paraprotein 7,5 g/l and free light chains (FLC) lambda our 1 year TRM and OS seems to be quite encouraging. 207 mg/l. Endomyocardial biopsy confirmed AL amyloidosis

S580 and trephine biopsy detected 10-20% infiltration of bone approach (A.Lengle, 2013): 1). The establishment of the marrow with clonal plasmatic cells.Final staging excluded therapeutic relationship, 2). Appeal to the present and the involvement of further target organs with AL amyloidosis. division of tasks, 3). Work on cognitive structures and incorrect Patient was indicated to peripheral blood stem cell harvest attribution, 4). Mobilization of personal resources and deter- (PBSC) and high-dose chemotherapy with autologous stem mine their position using specific existential-analytic methods cell transplantation (ASCT). - self-distancing, self-acceptance, finding personal position, 4). Materials (or patients) and methods: PBSC were mobilized Processing of failure sensations 5). Processing of guilt and with G-CSF (granulocyte colony-stimulating factor, filgrastim concretization of responsibility, 6). Work on relationships, 7). 10 mg/kg/day) alone. Even after escalation of G-CSF dose Deep therapy aimed at restoring the ability to experience the patient did not proceed to harvest. As a poor mobilizer fundamental values. patient received additional mobilization drug plerixafor Assessment of the dynamics of depressive disorders was (Mozobils, CXCR4 inhibitor) and proceed to aferesis with assessed clinically using a scale of depression Montgomery- successful harvest of stem cells (5,6.106/kg CD34 þ cells). Asberg (MADRS), Hospital Anxiety and Depression Scale Unfortunately within days after harvest, patient presented with (HADS) and the Clinical Global Impression (CGI) to assess the bilateral heart failure, NYHA III-IV, BMP 2100 ng/l, and pleural severity of illness (CGI-S) and improvement (CGI-I). and pericardial effusions. Patient was put urgently on Results: One measure of the success of this type of waiting list and within 2 months underwent uncomplicated therapy is to return the existential fundamental motivation heart transplantation. Histologic findings confirmed of life and the active component of personal action. Such a progression of amyloidosis in original heart. Posttransplant finding of personal position is followed by understanding the biopsy of a transplanted heart did not show any rejection or value of life. As a result of the study were obtained clinical signs of amyloidosis. (normalization or significant improvement in mood, normal- Six months following the heart transplantation, patient ization of sleep, appetite, new plans for the future) and underwent formerly planned ASCT with high dose chemother- psychometric (reduced scores on depression and anxiety, as apy melfalan 140 mg/qm. The ASCT was uncomplicated, only well as on the Clinical Global Impression scale) is evidence of mild toxicities and no serious perittransplant infections despite the effectiveness of existential approach to psychotherapy of ongoing immunosupressive treatment (prednison, tacrolimus) depression in cancer patients undergoing bone marrow were noticed. Two months after ASCT patient is clinically transplantation. stable, fully in outpatient settings, endomyocardial biopsy Conclusion: Existential psychotherapy is a quite effective negative in terms of amyloidosis and laboratory negative FLC treatment for neurotic level depression in patients undergoing confirm complete hematologic remission. bone marrow transplantation. Results: Disclosure of Interest: None declared. Conclusion: Published small patients cohorts after heart and bone marrow transplantation present the best results in AB065 patients reaching hematologic remission with 65% 5-year Is it possible to overcome the catastrophic complications survival. of Hematopoietic Stem Cell Transplantation (HSCT) There are a few case-report series reporting deterioration of using Plasma Exchange (PE)?: A Single Center’s heart failure in the course of PSBC stimulation with G-CSF. Our Experience case si the first report which describes heart failure after 1,* 1 1 1 stimulation of PBSC with G-CSF in combination with plerixafor M. K. Yuksel , G. Pekcan , E. Edibog˘lu , M. Bay , S. Civriz1, S. K. Toprak1, P. Topcuoglu1,O¨ . Arslan1,M.O¨ zcan1, to date. 1 1 1 1 1 References: Supported by Program development fields of M. Beksac , T. Demirer , H. Akan , N. Konuk , G. Gurman , O.˙ Ilhan1 science at Charles University PRVOUK - P27/LF1/1 and by grant 1 IGA Czech Ministry of Health - grant No. NT 11299-6/2010 Hematology Department and Stem Cell Transplantation Unit, Disclosure of Interest: None declared. Ankara University School of Medicine, Ankara, Turkey Introduction: Plasma Exchange (PE) is an effective AB064 therapy for TTP and desensitization for HSCT.However its Existential psychotherapy of depression in patients effectiveness for early complications of endothelial origin such undergoing bone marrow transplantation as venoocclusive disease (VOD) of liver, HSCT associated D. E. Vybornykh1,* on behalf of Research Group studying Mental thrombotic microangiopathy(TMA) and infectious complica- Disorders in Patients with Hematological Diseases tions such as sepsis and acute and chronic GVHD of liver is 1BMT, National Research Center for Hematology, Moscow, controversial. Our aim is to report effect of PE for this Russian Federation controversial issue. Materials (or patients) and methods: We conducted a Introduction: Depression in patients undergoing bone retrospective chart review of 40 patients who underwent an marrow transplantation is a serious problem in the treatment allogeneic HSCT between 2006-2014. The primary end point of blood diseases, because, in addition to difficult subjective was the effect of PE on the survival. experiences, they lead to a breach of compliance, which Results: Of the 40 patients 22 (55%) were male and 18(45%) promotes, in particular, a protocol violation of bone marrow were female. The mean age at transplantation was 34.7 (17- transplantation. From the perspective of the existential 67). Diagnosis of the patients were ALL n ¼ 6 (15%), AML approach, the painful depressive feelings makes to forget n ¼ 12 (30%), Biphenotypic Leukemia n ¼ 2 (5%),FAA n ¼ 2 about how important and necessary to appeal to the life and (5%), HL n ¼ 2 (5%),IMF n ¼ 3(7.5%), MDS n ¼ 6 (15%), MDS- life values, and the severity of depression determined by the AML n ¼ 2 (5%), NHL n ¼ 2 (5%), CML n ¼ 1(2.5%), CLL degree of violation of experience the value of life with the loss n ¼ 1(2.5%), MM n ¼ 1 (2.5%). All of the patients underwent of ability to perception of the value that previously gave it allogeneic HSCT. Of the 40 patients 23 (57.5%) were full match meaning. related donors, 16 (40%) were unrelated donors and one Aim. To conduct a study to examine the effectiveness of (2.5%) was haploidentical donor. PE indications were Sepsis existential psychotherapy of depression in patients under- n ¼ 13 (32.5%) VOD n ¼ 9 (22.5%) TMA n ¼ 6(15%), Acute going bone marrow transplantation. GVHD n ¼ 6 (15%),Chronic GVHD n ¼ 2 (5%), Pure Red Cell Materials (or patients) and methods: The study involved 21 Aplasia(PRA) n ¼ 1(2.5%), drug toxicity n ¼ 1(2.5%). Technical patients with various hematologic malignancies undergoing aspects of plasma exchange is shown in Table 1. Of the 40 bone marrow transplantation who were diagnosed with patients five of them (12.5%) were alive.The diagnosis of were neurotic level depression. Treatment was carried out with TMA n ¼ 2, Chronic liver GVHD n ¼ 1, PRA n ¼ 1, drug toxicity the use of methodological principles of existential-analytical n ¼ 1.

S581 Table Technical Aspects of Plasma Exchange and Laboratory increased risk for autoimmune disorders. No death occurred Results due to the above described manifestations. Conclusion: Autoimmune connective tissue manifestations Average Before PE After PE represent a rare but significant cause of morbidity after (Min-Max) allogeneic HCT. Due to the small number of patients we were not able to discern any association of the autoimmune Number of PE 4.5 (1-13) Hemoglobulin 8.7gr/dl 8.7 gr/dl manifestations with any variable. (gr/dl) (5,4-11.9) (6.2-11.4) Disclosure of Interest: None declared. PE duration 97.62 min (49-184) Volume 2740 ml Platelet(10^3/ 40 (1-287) 30.47 AB067 of exchanged (1150-4000) uL) (4-113) plasma Hepatic artery velocity in diagnosis of hepatic veno- Duration From 94.5 days Wbc(10^3/uL) 3.44 (0.1-23,7) 2.87 occlusive disease after bone marrow transplantation in Tx to PE (3-444) (0.1-20.3) children Duration from 132.5 days LDH (IU/L) 650(57-3223 1195 N. Kaya1,*, F. Erbey2, D. Atay2, A. Akc¸ay2,G.O¨ ztu¨rk2 Tx to death (10-699) (67-6223) 1 2 Duration from 48.8 days Total Biluribin 11.5 mg/dl 11.96 mg/ Radiology, Pediatric Hematology/Oncology & BMT Unit, PE to death (1-469) (mg/dl) (0.28-42.5) dl(0.24-33) Bahcelievler Medicalpark Hospital,˙ Istanbul, Turkey Introduction: Hepatic veno-occlusive disease (VOD) is a Conclusion: Plasma exchange is a good tool to remove toxic serious, early complication of bone marrow transplantation substances and replace healthy plasma. However, our institu- (BMT). The diagnosis of VOD is primarily based on achieve- tion experience with plasma exchange for posttransplantation ment of established clinical criteria. Several studies have complications especially for sepsis and VOD does not support focused on the role of ultrasound in the diagnosis of VOD. But this practice as a viable option but it may be a time serving there is no any study about role hepatic artery velocity in the bridge to other therapies such as chronic liver GVHD. diagnosis of VOD. Disclosure of Interest: None declared. Materials (or patients) and methods: Between April 2012 and June 2014 we obtained serial sonograms on 27 children who underwent HSCT prospectively. Ultrarosonography was AB066 performed before conditioning regimen and, after transplan- Development of Autoimmune Connective Tissue tation day þ 1, þ 7, þ 14, þ 28, and also performed at the manifestations after Hematopoietic Cell Transplantation time of the clinical and biological diagnosis of VOD. The In Children radiologist recorded, the direction, velocity and diameter M. AL-Darwish1,*, A. Al- Seraihy1, A. AL-Jefri1,2, A. ALAhmari1,2, portal vein, hepatic artery resistive index, hepatic artery I. Ghemlas1, A. Mohammed1, K. Siddiqui1, M. Ayas1,2 velocity, diameter of tree hepatic veins, mean velocity and 1Department of Pediatric Hematology Oncology, King Faisal flow pattern in the hepatic vein, splenik vein diameter and Specialist Hospital & Research Center, 2Al-Faisal University, velocity, thickness of the gallbladder wall, presence of ascites, Riyadh, Saudi Arabia liver and spleen size. The clinical diagnosis of VOD remaining according to modified Seattle criterias. Introduction: Hematopoietic cell transplantation (HCT) can be Results: Twenty seven patients (15 males and 12 females) with curative in many malignant and non-malignant disorders. The a median age of 10 (range 1-16 years) were included in this process however may potentially be associated with a study. Patient malignant and nonmalignant diseases were 14 multitude of complications. One of the rare sequelae of HCT and 13. myeloablative and nonmyeloablative conditioning is the development of autoimmune connective tissue mani- regimens were used in 24 and 3 patients. Clinical VOD was festations; the reported cases are mostly in adult patients. seen in 9 patients (group 1). There was no clinical diagnosed We, at King Faisal Specialist Hospital & Research Center VOD in 18 patients (group 2). The median hepatic artery (KFSHRC), have observed few such cases of in our pediatric velocity was 122±17 (95-153) and 44±18 (22-80) cm/sec. in patients after allogeneic HCT (o14 years), and report here our group 1 and group 2 respectively. the difference was observation. statistically significant (kruskal wallis test Po0,0001). Materials (or patients) and methods: Between January 2008 Conclusion: Measurement of hepatic artery velocity with and August 2014, 8 patients (2 males, 6 females) were doppler ultrasonography could predict diagnosis of VOD. identified to have developed autoimmune manifestations after References: 1. McCarville MB, Hoffer FA, Howard SC, Golou- allogeneic HCT; five patients had undergone HCT for non- beva O, Kauffman WM. Hepatic venoocclusive disease in malignant conditions (2 thalassemia, 2 Fanconi anemia, and children undergoing bonemarrow transplantation: usefulness one pure red cell aplasia), and 3 patients for malignant of sonographic findings. Pediatr Radiol. 2001 Feb;31(2):102-5. conditions (1 relapsed B cell acute lymphoblastic anemia 2. Herbetko J, Grigg AP, Buckley AR, Phillips GL Venoocclusive liver -ALL-, 1 early T cell ALL and 1 Familial Hemophagocytic disease after bone marrow transplantation: findings at duplex lymphohistiocytosis). sonography. AJR Am J Roentgenol. 1992 May;158(5):1001-5. Results: Mean age at HCT was 8.7 years (range, 0.78 -13.3 years) 3. Lassau N, Lecle`re J, Auperin A et al. Hepatic venoocclusive and mean time of follow up post HCT was 2.9 years (range, 0.3– disease after myeloablative treatment and bone marrow 6.2 years). The autoimmune manifestations developed after HCT transplantation: value of grayscale and Doppler US in 100 at a mean time of 8.2 months (range, 0.83–24 months). Five patients. Radiology. 1997 Aug;204(2):545-52. patients developed arthritis with positive autoimmune markers. Disclosure of Interest: None declared. Six patients developed serositis in the form of pericardial effusion with or without ascites and pleural effusion, two of which had isolated effusion without autoimmune markers or GVHD and one AB068 had pericardial effusion with GVHD and autoimmune pancyto- Polycythemia vera development after allogeneic stem cell penia. Additionally, 2 patients developed neurological manifesta- transplantation tions. All patients were treated with steroids; only one remains O. O. SAVAS1,1,*, S. Dagdas1, F. Ceran1,G.O¨ zet1, C. Sunu1, on therapy for Systemic lupus erythromatosis ( SLE) as he has G. Tokgoz1, M. Falay1 multiple risk factors including family history. 1Ankara Numune Research and Training Hospital, Ankara All patients engrafted 100%. Three patients are sex mis- Numune Research and Training Hospital, ANKARA, Turkey matched donor and one of them has one antigen mismatch and two cord blood. Each patient was found to have at least 2 Introduction: Allogeneic bone marrow transplant patients are HLA-alleles that have been reported to be associated with at risk for secondary malignancies as a late complication of

S582 treatment The secondary malignancies may present as a diffuse GS of bone which was prominent at knees. Again, his serious complication in patients with long life expectancy with bone marrow was in remission. The third patient was a 12 a wide range of clinical cases including EBV-associated B-cell years old male who had been transplanted with the diagnosis lymphoproliferative diseas, acute myeloid leukemia and of acute myeloid leukemia-M0. At the end of the first year after myelodisplastic syndromes. Polycythemia vera development allogeneic HSCT, he was diagnosed with bilateral testicular GS. has been rarely reported after transplantation in the literature. In this last case, bone marrow was also in remission as well as Therefore we planned to present two cases of polycythemia the other two cases. All three patients received salvage vera developing after transplantation chemotherapy after relapse and 2nd patient also received Materials (or patients) and methods: FIRST CASE: 25-year- radiotherapy. First and second patients received donor old man was admitted because of pancytopenia. Blood tests lymphocyte infusions as well, however, none of them achieved were unremarkable for pancytopenia etiology. The patient was remission. diagnosed with MDS and allogeneic stem cell transplantation Results: First and 2nd patients died 2 and 7.5 months after was performed from.fully compatible sibling donor. Hemato- relapse, respectively. The 3rd case is still alive without crit elevation was detected 3 years after transplantation. remission period after 4 months of GS diagnosis. (WBC:6300 NEU:3600 HB:17.9 HTC: 54 PLT:157000). Secondary Conclusion: Extramedullary relapse after HSCT is usually polycythemia causes were excluded in patient. JAK-2 muta- resistant to treatment because of the cumulative toxicity of tions were found to be positive and EPO values were within previously administered high dose . Addition- normal limits. We also studied the pre-transplant stored blood ally, high dose chemotherapy in this setting may cause sample of the patient and the donor. JAK-2 mutation were graft failure and suppresses graft versus leukemia effect. negative in both. Survival after salvage chemotherapy plus radiotherapy SECOND CASE: 40-year-old man was admitted because of has been reported to be shorter than 4 months in most gastrointestinal bleeding. As a result of investigations carried cases. Today, there is no still a standart treatment modality out the patient was diagnosed with AML M1-M2. Allogeneic and a determined guideline for treatment of patients stem cell transplantation was performed in second complete with GS developed after HSCT. In order to treat this particular remission from fully compatible sibling donor. Hematocrit fatal disease, further prospective and larger studies are elevation was detected 3 years after transplantation. needed. (WBC:6100 NEU:1900 HB:18 HCT:54 PLT:149000) Secondary References: 1. Ge L, Ye F, Mao X, et al. Extramedullary relapse polycythemia causes were excluded in patient. JAK-2 muta- of acute leukemia after allogeneic hematopoietic stem cell tions were found to be positive and EPO values were within transplantation: different characteristics between acute mye- normal limits. We also studied the pre-transplant stored blood logenous leukemia and acute lymphoblastic leukemia. Biol sample of the patient and the donor. JAK-2 mutation were Blood Marrow Transplant. 2014 Jul;20(7):1040-7. negative in both.Patients are still followed in outpatient clinic 2. Yoshihara S, Ando T, Ogawa H. Extramedullary relapse of without evidence of progression acute myeloid leukemia after allogeneic hematopoietic stem Results: Allogeneic stem cell transplantation involves the risk cell transplantation: an easily overlooked but significant of developing a variety of hematological malignancies in the pattern of relapse. Biol Blood Marrow Transplant. 2012 long term. Although presented in a number of publications in Dec;18(12):1800-7. the literature, especially the development of myeloprolifera- 3. Zago LB, Ladeia AA, Etchebehere RM, de Oliveira LR. tive disorders including polycythemia vera is very rare Testicular myeloid sarcoma: case report. Rev Bras Hematol compared to other hematologic malignancies. Essential Hemoter. 2013;35(1):68-70. thrombocytosis development is reported in a case report 16 Disclosure of Interest: None declared. years post-transplant. Our patients were evaluated as primary polycythemia because JAK-2 mutation were negative in AB070 both.pre-transplant stored blood sample of the patient and Secondary malignancies in beta-thalassemia major the donor. patients after allogeneic hemopoietic cell transplantation Conclusion: As a result of cases we present we emphasize the (HCT) necessity of long-term follow-up of patients for the develop- 1,* 1 1 1 ment of myeloproliferative disease after allogeneic stem cell P. Olioso , S. Santarone , E. Pennese , P. Bavaro , G. Papalinetti1, P. Di Bartolomeo1 transplantation. 1 Disclosure of Interest: None declared. Department of Hematology, Transfusion Medicine and Biotechnology, Pescara, Italy

AB069 Introduction: In 2008 we described the long-term results of Three Isolated Granulocytic Sarcoma Cases that Developed survival in patients with thalassemia major (TM) treated with After Allogeneic Stem Cell Transplantation HCT in our center1. The aim of the present study was to P. Is¸ık1,*,N.O¨ zbek1, S. Kirkiz1, Z. Avcı1, N. Yaralı1, G. Kabac¸am2, describe the development of secondary cancer in 3 patients B. Tunc¸1 with TM who were successfully treated with alloHCT. 1Pediatric Hematology, 2Radiology, Ankara Children’s Hematol- Materials (or patients) and methods: Between May 1983 and ogy and Oncology hospital, Ankara, Turkey April 2006, 115 patients with TM underwent alloHCT at our Center. Details about the patient’s and donor’s clinical Introduction: Granulocytic sarcoma (GS) after hematopoietic characteristics as well as the transplant protocol (conditioning stem cell transplantation (HSCT) is rarely seen and response to regimen, graft-versus-host disease (GvHD) prophylaxis, mar- treatment is limited. Disease itself has fatal prognosis and row harvesting and supportive therapy) have been described overall survival is between 4 to 9 months. Herein, 3 elsewhere2. Briefly, all patients were given busulfan (total dose, granulocytic sarcoma cases, that developed after HSCT, is 13-14 mg/Kg) and cyclophosphamide (total dose, 200 mg/Kg) presented. as conditioning therapy and cyclosporine and short-course Materials (or patients) and methods: Our first patient was a methotrexate as GvHD prophylaxis. The transplant-related 14 months old male patient. He had been transplanted with mortality at 1 year was 8.7%. The 20-year Kaplan-Meyer the diagnosis of acute myeloid leukemia-M7. Eight months estimate of disease-free survival was 85.7%. HCT was after allogeneic HSCT, he was admitted with profound successful with transfusion independence and definitive cure hypercalcemia. He was found to have GS on the upper sites in 99 patients. Among this patient population, 3 patients of both iliac crests without bone marrow relapse. The second developed secondary cancer. patient was a 12 years old male with chronic myeloid leukemia Results: Case 1: this female patient was transplanted in 1984 who had been transplanted after blastic phase. At the end of at 2 years from HLA identical sister. The clinical course was 4th month after allogeneic HSCT, he was diagnosed to have uneventful except for grade II acute GvHD treated with

S583 steroids. 11 years after transplant she presented with an months post transplant. Both of them have presented a GVHD enlargement of the parotid gland diagnosed as carcinoma. (gastrointestinal grade III steroid-resistant treated by INFLIXIMAB, She underwent chemotherapy but the course was rapidly and untreated cutaneous grade I). Several episodes of CMV progressive and the patient died 6 months later. infection occurred in both cases. The younger patient presented, Case 2: this male patient was transplanted in 1994 at 14 years 32 months after transplant, a refractory cytopenia with multi- from his HLA-identical sister. In the early post-transplant lineage dysplasia with normal cytogenetics. The second patient period he showed grade I acute GvHD with progression into presented, 14 months after transplant, a chronic myelomonocytic severe extensive chronic GvHD. In spite of several lines of leukemia with monosomy 7. When diagnosed, the chimerism of immunosuppressive therapy, chronic GvHD resulted in severe both patients was a 100% donor. pulmonary insufficiency. In 2006 at 12 years after transplant a Conclusion: With approximately 75 cases reported worldwide, carcinoma of the tongue was diagnosed and surgically the incidence of DCL keeps increasing, probably influenced by the removed. The patient received radiotherapy and remained advent of chimerism techniques. Several factors favoring the stable for 2 years when the disease progressed to the salivary occurrence of these complications have been reported, in glands and rapidly ended with the death. particulartheroleofthedefectivemarrowstroma,thefailureof Case 3: this female patient was transplanted in 1998 at 24 antitumor immunity and viral infections favored by the important years from her HLA-identical sister. The post-transplant course immunosuppression, preleukemic potential in the donor cell and was uneventful. She developed a carcinoma of the cervix 3 the residual effects of chemotherapy and irradiation. years following HCT. She underwent a successful surgical We report on two patients who received a cord blood treatment and is now alive and well 13 years later. transplant and who developed de novo MDS from the graft. Conclusion: The prevalence of secondary cancer in our study Growth factors which are necessary to full hematopoietic (3 cases out of 99 survivors, 3%) appears to be lower than that recovery and the oncogenic potential of some viruses (CMV) reported in patients receiving conventional therapy with seem be, in our experience, potential predisposing factors. regular transfusions and iron chelation (8 cases of 1073 DCL is a distinctive entity with intrinsic and extrinsic multi- patients, 8.5%)3. Anyway, the risk of malignancies after HCT factorial mechanisms, involving the cell of the hematopoietic underlines the need for life-long surveillance. Special attention niche. A better understanding of these mechanisms appears to should be made to the organ system at higher risk for be decisive to mitigate occurrences of these complications. secondary malignancies and continued research is required to Disclosure of Interest: None declared. quantify risk based not only on transplant factors, but on pre- transplant exposures, as well. AB072 References: 1. Di Bartolomeo P et al. Long-term results of Abstract Withdrawn survival in patients with thalassemia major treated with bone marrow transplantation. Am J Hematol 2008;83:528-530. 2. Di Bartolomeo P et al. The Pescara experience of allogeneic bone marrow transplantation in thalassemia. Bone Marrow AB073 Transplant 1997;19(Suppl.1):48-53. Usefulness of acute phase proteins and cytokines in 3. Borgna-Pignatti C et al. Survival and complications in monitoring of early engraftment after bone marrow patients with thalassemia major treated with transfusion and transplantation deferoxamine. Haematologica 2004;89(10):1187-1193. K. Gawronski1, P. K. Rzepecki1,* Disclosure of Interest: None declared. 1MILITARY INST. OF MEDICINE Dept. of Internal Medicine & Hematology, Warsaw, Poland

AB071 Introduction: The hematopoietic stem cells transplantation is Donor cell myelodysplastic syndrome after double an established therapeutic method- especially for patients umbilical cord blood transplantation: about two cases with neoplastic blood diseases. Many patients during the P. Arnautou1,*, J. Konopacki1,A.Se´got1, G. Dumas1, D. Bories2, transplant procedure have different long-term fever. Accord- B. Souleau1, J. V. Malfuson1 ing to clinical observations in our center, fever extends 1Department of hematology, Hoˆpital Percy, Clamart, 2Laboratory regeneration of hematopoietic system after transplantation. of molecular biology, Hoˆpital Henri Mondor, Creteil, France There could not be found publications that describe the problem for patients after transplantation. Introduction: Relapse after allogeneic stem cell transplanta- Materials (or patients) and methods: It has been attempted tion mostly originates in the initial malignant clone. Post to analyze the impact of fever on the process of hematopoietic allograft de novo acute myeloid leukemia (AML) or myelo- regeneration. It is clearly defined as: dysplastic syndrome (MDS) is a rare complication. Though ‘‘Engraftment is defined as the persistence of the 2 days these malignancies seem to be derived from the onco- segmented neutrophils (ANC)4 0.5 G / L or4 1.0 G / l for one genic transformation of recipient stem cells, the leukemogen- day. The engraftment of a platelet system is defined as sustained esis mechanisms of donor cell leukemia (DCL) remain platelet count4 20 G / l without platelet transfusion’’. unknown.We report the cases of two early de novo MDS An additional parameter indicating the regeneration of the occurred after umbilical cord blood (UCB) transplantation in hematopoietic system has been taken into account: the change in our department. the reticulocyte after transplantation. During the clinical study Materials (or patients) and methods: Two patients 38 (M) there were evaluated the acute phase proteins such as: and 60 (F) years old received double UCB transplantation procalcitonin, HS CRP, SAA, and some proinflammatory cytokines: between July 2011 and February 2013 for a core binding factor IL1Beta, IL4, IL6, IL12, TNFalfa, and antinflammatory IL1RA, IL10. AML in CR2 for him and a secondary AML with normal Results: There has been demonstrated a high correlation karyotype (RC1) for her. Non-myeloablative conditioning between the total number of days of fever, and the number of (CYCLOPHOSPHAMIDE 50 mg/kg, FLUDARABINE 150 mg/m2, days when the patient reached the number of ANC4 0,5 G/l, total body irradiation 4 Gy) and prophylaxis of GVHD ANC4 1,0 G/l. There have also been observed a high (Cyclosporin and Mycophenolate Mofetil) were identical in correlation between the total number of fever days, and the the two cases. number of days when the patient reached the of value PLT4 Results: In both cases, the allograft was complicated by a late 20G/l and PLT4 50G / l. There has been a significant hematopoietic reconstitution (neutophil count4500/mm3 at D39 difference in results between patients with fever and patients and D38 and platelets count420,000 / mm3 at D292 and NA, without fever regarding levels of procalcitonin, HS CRP, SAA respectively) and an extended hematopoiesis support by triple and less cytokines – IL1RA, IL6. In each case, the growth factors therapy was necessary (GCSF, EPO and TPO average level observed in patients with fever was higher than agonist). Chimerism was a 100% donor for each patients from 3 the average level for patients without fever.

S584 Conclusion: The results show that all the tested acute phase Conclusion: In this report, we describe the experience of proteins: procalcitonin, HS CRP, SAA and cytokines IL1RA, IL6, therapeutic DF for VOD after ASCT. The incidence of VOD in correlate with the time of the hematopoietic system regenera- patients who underwent ASCT at our centre was 9.1%. The tion. The level of these factors increases in patients febrile for mortality rate of severe VOD is similar to literature. DF is the two days or longer. In our study we have demonstrated the only agent approved for use in severe hepatic VOD following relation between fevers in the early period after transplantation, HSCT and represents a useful advance in the treatment of this and regeneration of the hematopoietic system. The high value condition. of procalcitonin can have a direct effect on hematopoietic Disclosure of Interest: None declared. recovery delay. Unfortunately we were unable to determine a specific value or range of values at which hematopoietic AB075 recovery time is longer. This application is a novel observation, Effect of different conditioning regimens on nutrition and because so far no one has proved the correlation of blood parameters on patients undergoing autologous procalcitonin to achieve appropriate rates of engraftment. stem cell transplantation Disclosure of Interest: None declared. V. Kesik1,*, E. Atas1, C. Akyuz2 1Department of Pediatric Oncology, Gulhane Military Medical 2 AB074 Academy, Department of Pediatric Oncology, Hacettepe Defibrotide in the treatment of hepatic veno-oclusive University, Ankara, Turkey disease after allogeneic stem cell transplantation: A retrospective single center experience Introduction: Nutrition plays an essential role in autologous stem cell transplantation (ASCT) with primary effects on P. Patir1,*, N. Soyer1, M. Tombuloglu1, G. Saydam1, F. Vural1 1 healing mucositis and fighting infections. Patients undergoing Department of Hematology, Ege University Medical Faculty, transplantation process are at high nutritional risk and should IZMIR, Turkey undergo careful nutritional assessment. We evaluated the effect of different conditioning regimens (CR) on nutrition, Introduction: Veno-occlusive disease (VOD) of the liver is a metabolism and serum parameters in children with various complication observed particularly in patients undergoing cancers. allogeneic stem cell transplantation (ASCT). Defibrotide (DF) is Materials (or patients) and methods: As a standard care, we the sodium salt of a single-stranded polydeoxyribonucleotide use total parenteral nutrition (TPN) for patients with no oral with anti-ischemic, anti-inflammatory and thrombolytic effect feeding for 5 days. We evaluated the weight, duration of TPN, without significant anticoagulant property. We evaluated the lymphocyte count at post transplant day 15 and 30, serum efficacy of DF in patients developing hepatic VOD after ASCT albumin, GGT, ALP, AST, ALT, total/direct bilirubin values of 48 in a retrospective analysis. patients at the beginning of the transplant and at discharge, Materials (or patients) and methods: One hundred and retrospectively. The effect of CEM, Bu-Mel, and BEAM seventy five ASCT were performed in the Ege University conditioning regimens on these parameters were analyzed. Bone Marrow and Stem Cell Transplantation Unit from Kruskal-Wallis and Mann-Whitney-U test were used for January 2008 to September 2014. Sixteen (13 male and 3 analysis. female) out of 175 patients were diagnosed with VOD Results: The mean age was 9.1 years (range 10 months-18 based on the Seattle criteria retrospectively. Median age years) and male/female ratio was 15/33 ¼ 0.45. There were 13 was 51 (18- 59). Thirteen patients received myeloablative subjects in BEAM, 18 subjects in Bu-Mel, and 17 subjects in (busulfan- cyclophosphamide) and 3 patients received CEM. Mean duration times of TPN in BEAM, Bu-Mel, and CEM reduced intensity conditioning regimen (busulfan-fludarabine- were 14.3, 11.9, and 18.8 days, respectively (P ¼ 0.083). There cyclophosphamide). were differences between groups of CR according to mean Results: Severity, treatment results of patients with baseline and discharge value of subjects GGT (% change) VOD were illustrated in Table 1. A total of 16 patients (P ¼ 0.001) and lymphocyte count on day þ 15 (P ¼ 0.008) and out of 175 allo-transplanted patients (9.1%) with median age day þ 30 (P ¼ 0.029). Statistically differences were found of 45.3 years (range;18-59 years) were diagnosed as hepatic between BEAM and Bu-Mel, and between Bu-Mel and CEM VOD. 12 patients were treated with DF but DF was not according to GGT, between Bu-Mel and CEM, and between provided for the treatment of other 4 because of the insurance BEAM and CEM according to post HSCT lymphocyte count on procedures. Seven patients (44%) met the risk criteria for day þ 15, between BEAM and Bu-Mel, and between BEAM and severe, 5 patients (31%) moderate, and 4 patients (25%) mild CEM according to post HSCT lymphocyte count on day þ 30 disease. The median duration of DF treatment was 10 days. In (Mean differences of % GGT; BEAM ¼ 47.6 vs Bu-Mel ¼ 11.1, all, 6 patients (50%) achieved complete response. CR rate in P ¼ 0.009; Bu-Mel ¼ 11.1 vs CEM ¼ 187.1, P 0.0001, patients with severe, moderate and mild diseases were 17%, o Mean differences of lymphocyte count on post HSCT þ 15; 75% and 100% respectively. The mortality rate of severe VOD Bu-Mel ¼ 0,38 mm3 vs CEM ¼ 0.16;P ¼ 0.028, BEAM ¼ 0.71 vs was 83%. CEM ¼ 0.16, P ¼ 0.004, Mean differences of lymphocyte count on post HSCT þ 30, BEAM ¼ 1.4 vs Bu-Mel ¼ 0.93, Table 1. Characteristics and treatment of patients with VOD P ¼ 0.016, BEAM ¼ 1.4 vs CEM ¼ 0.87, P ¼ 0.03). Although statistical differences were not found, body weights of Onset Grade of Duration of Dose of DF Delay to start Outcome subjects were decreased according to CR (Differences body of VOD DF therapy therapy (mg/ DF after weight %; BEAM ¼ -6.5, Bu-Mel ¼ -4.9, CEM ¼ -0.54, VOD kg/day) diagnosis P ¼ 0.201). 13 Moderate 14 25 17 CR Conclusion: As conclusion, CEM, BEAM and Bu-Mel made 11 Severe 23 25 0 NR significant chemotherapy related anorexia (18.8, 14.3, 11.9 35 Severe 7 25 3 NR days respectively). CEM and fewer BEAM signi- 23 Severe 4 25 1 NR ficantly increased GGT that was correlated with the 7 Moderate 12 25 2 CR risk of cholestasis. CEM significantly reduced lymphocyte 9 Severe 15 25 7 CR 10 Mild 13 25 2 CR levels than Bu-Mel that increase predisposition to infections. 7 Severe 9 25 3 NR TPN duration was directly correlated with GGT level. Investiga- 2 Severe 1 25 13 NR tion the role of nutritional support on infections and 3 Moderate 2 25 5 NR engraftment status in large series with further studies may 8 Moderate 18 25 1 CR give more information. 5 Mild 13 25 2 CR Disclosure of Interest: None declared.

S585 [AB075]

AB076 patients gave written consent. The inclusion criteria were Haematopoietic Stem Cell Transplant-related Oral age418 years old, WHO grade 0 at baseline, ability to read, Mucositis and the use of a patient self-assessment scale in and not receiving investigational agent(s) for mucositis. an Asian population Exclusion criteria were subjects receiving non-myeloablative W. Shu Ping1,*, C. Kian Meng1, M. Z. Zakaria1, G. Ai Sim2, conditioning regimen. The WHO and OMDQ (Q2) was defined C. Su Kien2, F. Islahudin3 on behalf of Malaysia Mucositis as mucositis (Grade Z1) and severe mucositis (Grade 3–4). Research Group Results: A total of 23 patients and 464 days of hospital stay 1Hospital Ampang, Selangor, 2Hospital Pulau Pinang, Pulau were analysed. 14 (60.9%) patients experienced OM by both Pinang, 3Universiti Kebangsaan Malaysia, Kuala Lumpur, scales which is lower than published literature, suggesting Malaysia inter-ethnic differences in chemotherapy drug handling. According to WHO scales, only 35% were classified "severe" Introduction: Oral mucositis (OM), a frequent complication of as the highest grade of mucositis experienced, in contrast to Haematopoietic Stem Cell Transplant (HSCT), is associated with 71% according to OMDQ scales. Patient self-report was able to significantly worse clinical and economic outcomes. OM may capture an earlier onset of OM by 3.1±SD 2.8 days (P ¼ 0.024), differ across populations due to variances in drug metabolism. where OM onset by WHO score was mean 12.5 days (SD±2.7, Local data on OM in HSCT is limited. Furthermore, guidelines range: 9–19 days) and by OMDQ was mean 9.4 days (SD±4.1, recommends that patient self-report forms part of OM range: 1–17 days). This may reflect patients’ under-reporting of assessment, which is currently not extensively used or studied mild pain. OM typically lasted for a mean duration of 6.3 days in this population. (SD±1.97, range: 3–9 days) by WHO scores and 9.86 days Materials (or patients) and methods: In this prospective (SD±4.5, 1–22 days) by patient’s report. Several factors observational study across two HSCT centres and involving a associated with mucositis are presented in Table 1. The length universal sampling of patients consecutively admitted, World of hospital stay was found to be an average of 20.1 days Health Organization (WHO) oral toxicity scores were obtained (SD±2.8, range: 16–26 days). OM was significantly associated during hospital stay. Patients also recorded in a daily diary with 2.6±SD 4.1 additional days of hospital stay (P ¼ 0.023) containing the Oral Mucositis Daily Questionnaire (OMDQ). and 2.5±SD 3.8 additional days of systemic analgesic use The study was approved by the Ethics Committee, and all (P ¼ 0.03). A total of 6 (26%) of patients required strong

[AB076]

S586 opioids. The data indicated that pain control deteriorated as Conclusion: This study showed that the sequence of Cy and severity of OM increased, with most of the pain episodes TBI does not impact transplant outcomes for children with requiring strong opioids only achieving moderate or no relief acute leukemia. However, the results suggest that acute (Po0.001). This perhaps suggests differences in cultural nausea and vomiting and chronic GVHD are possibly lower conception on the use pain . with Cy/TBI sequence, although further evaluation is war- Conclusion: In conclusion, this data provides important ranted to confirm these findings. preliminary insight into the scope of mucositis in the Disclosure of Interest: None declared. Malaysian transplant setting. Mucositis pain continues to be a clinical problem in this type of population and the use of AB078 self-reported scales has potential advantages in the clinical The effect of busulfan dosing in fludarabine based management of OM, which could lead to earlier changes in regimens on clinical outcomes in patients who were therapy and may prove useful if mucositis reporting indepen- undergoing high risk hematopoietic stem cell dent of clinical visits is required. transplantation References: Bensinger W, et al. J Natl Compr Cancer Netw. D. S. Hong1,*, S. K. Park1, S. H. Kim1, J. Yun1, H. J. Kim1, K. H. Kim2, 2008. 6(11): 1–21 2 1 Loh M et al. Pharmacogenomics J. 2013; 13(5): 423-429 J. H. Won , C. K. Kim 1Hematology, SOONCHUNHYANG UNIVERSITY HOSPITAL, Quinn B, et al. Eu J Cancer 2008. 44(1): 61-72 2 Chen CH, et al. Palliat Med. 2012; 26(3): 206 -221 Bucheon, Hematology, SOONCHUNHYANG UNIVERSITY Breuer B, et al. J Clin Oncol. 2011; 29:4769-4775. HOSPITAL, Seoul, Korea, Republic Of Disclosure of Interest: W. Shu Ping Funding from: National Institute of Health, Ministry of Health, C. Kian Meng: None Introduction: The reduced conditioning regimen consisting of declared, M. Z. Zakaria: None declared, G. Ai Sim: None reduced dose bvsulfan and fludarabine, and ATG are widely declared, C. Su Kien: None declared, F. Islahudin: None used and less toxic, induced sufficient immunosuppression in declared. the host to allow donor hematopoietic cells to consistently engraft across the HLA barrier even haplotype barrier. However, dose intensity across reduced intensity conditioning regimens vary and may affect treatment outcomes. In this retrospective analysis, we investigated the effect of Conditioning regimen intravenous busulfan dosing (Bu2, 6.4 mg/kg of busulfan versus MA, 12.8 mg/kg of busulfan) in fludarabine based regimens that combined fludarabine (160 mg/m2 -180 mg/m2) and busulfan on clinical outcomes in patients who were AB077 undergoing high risk hematopoietic stem cell transplantation. The effect of Administration Order of Cyclophosphamide Materials (or patients) and methods: A total of 80 patients and Total Body Irradiation on Outcome of Allogeniec who underwent peripheral blood stem cell transplantation Hematopoietic Stem Cell Transplantation for Children with from unrelated donor or mismatched familial donor including Acute Leukemia haploidentical donor with fludarabine based regimens. A. A. Hussein1,*, R. Najjar2, K. Salami1, A. Haroun1, R. Farraj2, Results: Of the 80 patients, 25 patients received busulfan of A.-H. Al-Zaben1 reduced intensity (Bu2) and 1Bone Marrow and Stem Cell Transplantation, 2Department of 55 patients received myeloablative busulfan (MA). Patients Pharmacy, King Hussein Cancer Center, Amman, Jordan had similar characteristics. Median follow-up for survivors was 41 months. The day þ 100 cumulative incidence rate of Introduction: The effect of sequencing Cyclophosphamide grades II to IV acute GVHD (36% for Bu2, 44% for MA, (Cy, 120 mg/kg) mg/kg and myeloablative doses of Total Body respectively) was not different. The 2-year cumulative Irradiation (TBI, 1200cGY) in the conditioning regimen on incidence of chronic GVHD was not significantly different acute toxicities and outcome of HSCT for children with between Bu2 and MA. The nonrelapse mortality rates within leukemia is still unknown. 100 days after transplantation were similar for Bu2 and MA Materials (or patients) and methods: The sequence of Cy (16% versus 14.5%, respectively). Two-year overall survival and TBI was evaluated in all consecutive children with acute were also similar between Bu2 and MAu2 (57.2% versus 62.2%, leukemia who received HSCT from fully HLA-matched related respectively; P ¼ .99). donors at King Hussein Cancer Center in Jordan from January Conclusion: The dose of busulfan (6.4 mg/kg versus 12.8 mg/ 2005 to March 2014. All patients received the same GVHD and kg) is not associated with significant differences in clinical antimicrobial prophylaxis. outcomes for patients undergoing peripheral blood stem cell Results: A total of 60 pediatric patients were identified, with transplantation from alternative donors after conditioning median age of 10 year (4-20). 62% were males, 38 (63%) with busulfan and fludarabine. patients had ALL and 22 (37%) AML. Forty-five patients Disclosure of Interest: None declared. received TBI/Cy and 15 received Cy/TBI conditioning regimen. The two groups were controlled for primary disease (ALL, AML), disease status at transplant (CR1, 2, 3 and more) and stem cell AB079 source (Bone marrow or peripheral blood). The acute nausea BUSLERA R (BIEM drug company, Ankara, TURKEY); and vomiting was less in the Cy/TBI group (28.6% vs. 60.9%, generic busuphan is safe for conditioning in allogeneic Po0.001). However, there was no difference in the incidence of hematopoietic stem cell transplantation, single center delayed N/V (P ¼ 0.56), acute significant liver function abnorm- experience alities (P ¼ 0.23); acute gastrointestinal toxicity (P ¼ 0.75) E. Soydan1,*, A. Gokmen1, M. Kurdal1, D. Ozbek1, O. Ilhan2, including mucositis (P ¼ 0.23), hemorrhagic cystitis (P ¼ 0.66) O. Arslan 2, M. Ozcan2 and CMV reactivation (P ¼ 0.11) between the 2 groups. There 1Medicana Hospital, 2Ankara University Medical School, Ankara, was no significant difference in the incidence of grades II-IV Turkey acute GVHD (35.7% vs. 30.4%, P ¼ 0.25). While the incidence of chronic GVHD was lower in the CY/TBI group (7.1% vs. 37%, Introduction: Busulphan is one of major drugs of conditioning Po0.001). The was no significant difference between the 2 in allogeneic hematopoietic stem cell transplantation (AHSCT) groups on the rate of 100 days transplant-related mortality (0% both in ablative or reduced intensity setting. We used vs. 2.1%,), 3-year leukemia relapse (42.9% vs.% 43. P ¼ 0.9), BUSILVEXR (Pierre Fabre drug company,˙ Istanbul, TURKEY) leukemia-free survival (53.6% vs. 57.9%, P ¼ 0.77), and overall for 48 patients since the begining of our allogeneic transplant survival (69.9% vs. 54.5%, P ¼ 0.001). activity. Since September 2014, we are using BUSLERA for

S587 ablative and reduced intensity conditioning at same doses of the nine patients had different degrees of liver damage already we used for busilvex (0,8 mg/kg x4/day 4 days and 2 prior to transplantation. The donors were identical siblings (3), days, respectively). Adjusted ideal body weight was used for matched related donors (4) or haploidentical parents (2). dose calculation. We retrospectively overview the side effects The source of stem cell was bone marrow in 7 and for the first eight patients. peripheral blood in 2 of the patients. Conditioning regimen Materials (or patients) and methods: The charactristics of consisted of treosulfan /fludarabine in 8 patients and one of the patients were; seven acute leukemia, six in first remission them also received Thiotepa. The last patient received one in second remission and one chronic phase chronic Treosulfan/Cyclophosfamide. GvHD prophylaxis consisted of myeloid leukemia refractory to all available tyrosine kinase Cyclosporin-A in most patients. inhibitors. Five men, three women, ages between 21-62 years. Results: Severe exfoliative dermatitits was developed Four patients were given ablative conditioning and the other in two of the patients in early posttransplant period. VOD were prepared with reduced intensity conditioning. Two of the was detected in one patient and 4 patients developed donors were metched unrelated, all the others were HLA aGVHD grade II. Median day of engraftment was þ 11, idantical siblings. ATG was added to conditioning in match ranging þ 11 to þ 19. One patient with NIK defect is unrelated donors at a dose of 45-60 mg/kg according to the died at þ 3d with sepsis. Eight of the 9 transplanted patients presence of mismatch. Sorror comorbidity index was low in all are alive with complete clinical and immunological recovery patients. after HSCT. Results: We did not detect any severe adverse event Conclusion: These results indicate that the combination related to buslera infusion. Two out of eight patients had of treosulfan and fludarabine is effective in establishing grade 1 cardiac and 3 had grade 1 gastrointestinal side early donor engraftment with low toxicity and effects. Grade 3-4 mucositis were seen in four patients. improved survival in patients with severe combined/combined Engraftment kinetix were mean 13,3 (0-22days) and 9,5 days PIDs. (0-23days) for neutrophile and trombocyte recovery, Disclosure of Interest: None declared. repectively. Mean 3,2 (0-10) and 6,38 (0-12) u¨nits of packed red blood cells and single unit apheresis trombocytes were AB081 infused, respectively. Impact of Rituximab in FEAM conditioning regimen Conclusion: We are planning to make a retrocpective case for non Hodgkin Lymphoma match analysis when we have enough patients for statistical 1,* analysis. Till then we may say that buslera is a effective and F. Mazziotta on behalf of Francesco Mazziotta, Enrico Orciuolo, safe generic drug for conditioning during AHSCT. Valentina Bozzoli, Gabriele Buda, Anna Rita Messa, Lorenzo References: 1– Gupta T1, Kannan S, Dantkale V, Laskar Iovino, Pasquale Forese, Francesco Ghio, Michela Dargenio, Rossella Matera, Nicola Di Renzo and Mario Petrini S.Cyclophosphamide plus total body irradiation compared 1 with busulfan plus cyclophosphamide as a conditioning Pisa and Lecce Hospital, Pisa, Italy regimen prior to hematopoietic stem cell transplantation in patients with leukemia: a systematic review and meta-analysis. Introduction: Impact of Rituximab in FEAM conditioning Hematol Oncol Stem Cell Ther. 2011;4(1):17-29. 2-Nath CE1, regimen for non Hodgkin Lymphoma Shaw PJ.Busulphan in blood and marrow transplantation: Authors: Francesco Mazziotta1, Enrico Orciuolo, Valentina Bozzoli, dose, route, frequency and role of therapeutic drug monitor- Gabriele Buda, Anna Rita Messa, Lorenzo Iovino, Pasquale Forese, ing.Curr Clin Pharmacol. 2007 Jan;2(1):75-91. 3- Ferry C1, Socie´ Francesco Ghio, Michela Dargenio, Rossella Matera, Nicola Di G.Busulfan-cyclophosphamide versus total body irradiation- RenzoandMarioPetrini. cyclophosphamide as preparative regimen before allogeneic Introduction hematopoietic stem cell transplantation for acute myeloid Rituximab is a monoclonal antibody against CD20 which has leukemia: what have we learned?Exp Hematol. 2003 completely changed the natural history of CD20 positive Dec;31(12):1182-6. lymphomas. Disclosure of Interest: None declared. Its effectiveness when combined with chemotherapy regimens is amply demonstrated, so that has now become the standard of care in the treatment of non Hodgkin lymphomas (NHL). AB080 Despite rituximab being widely adopted in the context of Treosulfan Based Conditioning Regimens by Alogeneic conditioning regimens, limited data are available to guide the Stem Cell Transplantation in Patients with Primary management of the antibody in this field. Immune Deficiencies (PID) The aim of this study is to clarify the role of Rituximab S. Haskologlu1, F. Dogu1,*, C. Odek2, A. Yaman2, D. Guloglu1, added to fotemustine, etoposide, cytarabine and melphalan B. Kucukersan1, F. Arpacı3, T. Kendirli2, A. Ikinciogulları1 (R-FEAM conditioning) in CD20 positive non-Hodgkin 1Department of Pediatric Immunology-Allergy, BMT Unit, Lymphomas. 2Department of Pediatric˙ Intensive Care Unit, Ankara University, Materials (or patients) and methods: School of Medicine, 3Department of Medical Oncology, Gulhane Patients and methods Military Medical Academy, Ankara, Turkey Two centers contributed to this survey. We collected 61 NHL CD20 þ , 21 of them underwent R-FEAM conditioning, the Introduction: Treosulfan (treo) is an alkylating agent with a other 40 were treated with FEAM. low acute toxicity profile that is increasingly used in Patient characteristics were organized as in the table below. hematopoietic stem cell transplantation, mainly in non- All patients were evaluated for stage at diagnosis, response prior to malignant diseases. Here we present the outcomes of and after transplants, median times to engraftment and toxicities. nine patients with severe combined/combined PIDs under- Results: Results went HSCT using different treosulfan-based conditioning No differences were found between groups in terms of toxcitiy regimens. and times to engraftment. Low grade mucosities, chemother- Materials (or patients) and methods: MATERIALS AND apy-induced nausea and vomiting and diarrhea were the main METHODS: Nine patients with SCID/CID who underwent side effects recorded. Median time to neutrophil (4 500/mcl) allogeneic HSCT after treosulfan based conditioning regimens recovery where 10 days for FEAM and 9,5 days for R-FEAM were evaluated retrospectively. Two patients had SCID (T-B- regimen (P not significant); as for platelets recovery (420000/ NK þ ), 7 patients had CID (MHC Class II deficiency (2), CD40 L mcl) the median times were 12 and 9 days (P not significant), deficiency (1), DOCK 8 deficiecy (1), CD3 zeta chain defect (1), respectively. Omenn Syndrome(1), NIK defect(1). The median age at There was no difference, regarding overall survival (OS) and diagnosis was 4 months (range 1-36 months). CMV and progression free survival (PFS) (P not significant), in the R þ Human parainfluenza type III were leading viruses and seven and R – groups.

S588 A different trend in overall survival and progression free prophylaxis with cyclosporine (CyA) and methotrexate (Mtx). survival was observed between two subsets of patients: early In all of the patients full engraftment was achieved. Discharge (stage I-II) and advanced (stage III-IV) disease at diagnosis, in days were þ 32 and þ 38.day respectively. There were no favour of the early one, independently of Rituximab use or not. major toxicity and no GVHD. They are alive and transfusion Again, response status before transplant impact on OS and independent with full donor type engraftment and chimerism PFS, independently of Rituximab use: patients who achieved a (Table 1). deeper response had longer overall and progression free Conclusion: Treosulfan-based myeloablation might be a survival.Transplant related mortality in FEAM and R-FEAM was suitable elective application in pediatric patients or in those respectively 7,5% and 14,3%. with poor performance status and/or organ dysfunction, who Conclusion: Discussion have a high risk of life-threatening complications if busulfan is Rituximab has represented an important turning point in the employed. Future prospective, randomized trials comparing treatment of lymphomas. However in the context of a this treosulfan-based and the busulfan-based regimens are conditioning regimen, in view of a transplant procedure, the warranted. use of rituximab does not appear to have such an impact as in Disclosure of Interest: None declared. the previous lines of chemotherapy. Disclosure of Interest: None declared. AB083 Successful Hematopoietic Stem Cell Transplantation AB082 of a Child with Treosulfan Based Regimen, Who Had Treosulfan Based Preparative Regimens for Allogeneic Therapy-Related Acute Myeloid Leukemia and a Rare Hematopoietic Stem Cell Transplantation in Thalassemia Tanslocation: t (11;11) Major: Report of Two Pediatric Cases From a Single-Centre O. Gu¨rsel1, I. Eker1,*, C. Bozkurt2, S. Bozkurt3, O. Babacan1, I. Eker1,*,O.Gu¨rsel1, O. Babacan1,A.E.Ku¨rekc¸i1 A. E. Ku¨rekc¸i1 1Pediatric Hematology, Gulhane Military Medical Faculty, Ankara, 1Pediatric Hematology, Gulhane Military Medical Faculty, Turkey 2Pediatric Hematology and Oncology, Dr. Sami Ulus Children Hospital, 3Basic Oncology, Insitute of Oncology, Hacettepe Introduction: Hematopoietic stem cell transplantation (HSCT) University, Ankara, Turkey is the definite treatment for patients with thalassemia major. A busulfan (Bu) and cyclophosphamide (Cy) based regimen has Introduction: Childhood and adult cancer survivors have an been the standard myeloablative chemotherapy, but it is increased risk of developing second and subsequent cancers associated with higher treatment-related toxicity, particularly that varies by host factors and primary cancer therapy. The in patients classified as high risk by the Pesaro criteria. survival of patients with secondary-AML (s-AML) is generally Treosulfan-based conditioning regimens have been found to shorter than for those with de novo AML within the same be equally effective and less toxic. Here we report two cytogenetic risk group. In this article, we present a successful pediatric thalassemia major patients, who underwent HSCT hematopoietic stem cell transplantation (HSCT) of a new child with treosulfan / thiotepa / fludarabine ( treo / thio / flu) based case who has s-AML with a rare translocation; t(11;11), conditioning regimens, using grafts from their siblings, who following chemotherapy for ALL. are selected before implantation to be both unaffected and Materials (or patients) and methods: A 12 year old girl HLA-matched donors. hospitalized in our department for HSCT with the diagnosis of Materials (or patients) and methods: Two pediatric 10 years s-AML in complete remission (CR2). From her history it is old and 16 years old male patients with thalassemia major,, learned that she was diagnosed as pre-B-cell-ALL at 7,5 year- transplanted at our centre between June 2012 and July 2013 old and complete hematologic response had been achieved with treo / thio / flu based conditioning regimens, using grafts with St. Jude XIII protocol. The protocol was consisted a total from their siblings, who are selected before implantation to be 14,4 g/m2 of etoposide and 180.000 U/m2 of L-asparaginase. both unaffected and HLA-matched donors. They had been Conventional cytogenetic evaluation at the beginning and at treated with regular RBC transfusions and chelation therapy the end of the protocol had revealed a normal karyotype. She and both of them were assigned to class II, according to sustained her first CR for 2 years, after the termination of classification proposed by Pesaro group. Stem cell source in protocol, untill her first hematological relapse as AML-M4. both of the transplantations were bone marrow (BM) and Lumbar puncture at the time of relapse was also positive for umblical cord blood (UC). All of the patients received a myeloblast. Conventional cytogenetic evaluation of 20 meta- cumulative treo dose of 42 g/m2, a cumulative thiotepa dose phases of bone marrow leukemic cells at the relapse showed of 8 mg/kg and a cumulative fludarabine dose of 160 mg/m2. the following karyotype profile: 46,XY, t(11;11) in all of the Both of the patients received graft-versus-host disease (GVHD) cells. He had failed the remission induction therapy with AML prophylaxis with cyclosporine (CyA) and methotrexate (Mtx). BFM-87 protocol, but complete hematologic and central nerve Results: In both of the patients full engraftment was achieved system response has been achieved with AML BFM-2004 with the values of 3,2 106/kg and 3,5 105/kg respectively. protocol. She underwent HSCT from her full matched sister at All patients received graft-versus-host disease (GVHD) 12 years of age. She received myeloablative regimen

[AB082]

S589 consisting of treosulfan (36 mg/kg), cyclophosphamide fludarabine and Bu (n ¼ 18; 85.7%) and BM was the main (120 mg/kg) and etoposide (30 mg/m2). She also received source. Immunosuppression was based on FK-506/MTX (90.5% methotrexate and cyclosporine for graft-versus-host disease of patients). The median of AUC in 24 hours was 5690 (GvHD) prophylaxis. A total of 8,6 106 / kg CD34 þ cells was mMol.min (3539.6- 8881.8 mMol.min). The cumulative incidence infused. (CI) of sinusoidal obstructive syndrome (SOS) in the retro- Results: She achieved full hematopoietic engraftment with spective group and IV Bu were 9.5% for both, while in the the time for neutrophil, platelet, and erythrocyte recovery group oral Bu it was at 19% (P ¼ 0.566). The CI of acute graft- being 20 days, 25 days and 41 days respectively. She has not versus-host disease (aGVHD) at D þ 100 was 38.1% in the developed acute or chronic. The patient is at one year after retrospective group, 40.6% in oral Bu and 42.9% in IV Bu. HSCT and she is in complete hematological and molecular Chronic GVHD was 13.6% in oral Bu, 34% in IV Bu and 42.9% in remission with full donor-type chimerism. the retrospective group (P ¼ 0.142). The CI of relapse at Conclusion: The cases of s-AML can be divided in two groups: D þ 100 was 19% in IV Bu, 4.8% in the retrospective group and patients who received alkylating agents and who received oral Bu did not have this event. The CI of death at D þ 100 was epipodophyllotoxins like etoposide. The cases classified in last 34.9% in group oral Bu, 9.5% in IV Bu and 14.3% in the group are rarely preceded by a preleukemic phase, show retrospective (P ¼ 0.102). The CI of relapse at 1.5 years was abnormalities of the long arm of chromose 11 (11q23 region), 35.8% in the IV Bu, 34.8% in oral Bu and 14.3% in the involve monoblasts or myelomonoblasts and a short latency retrospective group. The CI of death at 1.5 years was 9.5% in time (1-3 years). Risk increases especially with high (more than group IV Bu, 53.5% in oral Bu and 34.3% in the retrospective 5 mg/m2) doses. L-asparaginase can also potentiate the (P ¼ 0.015). Among patients who died until D þ 100, the leukemogenic effect of etoposide. An improved understand- median of AUC was 5732 (5578.5- 6818.5 mMol.min) during the ing of therapy-related secondary malignancies can guide conditioning for IV Bu and (3448-8212 mMol.min) for oral Bu. alterations in regimens to minimize cytotoxic treatment The range between the AUC was large and there was no exposure. Besides effective screening methods and preventive correlation with patients who died. strategies should be developed to risk for second primary Conclusion: Relapse rates at D þ 100 were greater in group IV cancers. Treosulfan based conditioning regimens can be Bu (19%) that had more patients with active disease and up to considered for the HSCT of these patients. CR2. On the other hand, the death rate at 1.5 years was higher Disclosure of Interest: None declared. in the group oral Bu (P ¼ 0.015). SOS had an impact on mortality at D þ 100 after HSCT (p o0.005), with lower rate of AB084 AUC in oral Bu when compared to IV Bu. The clinical and therapeutic drug monitoring of oral and Disclosure of Interest: None declared. intravenous busulfan in patients with acute leukemia that underwent to stem cell transplantation with a test dose of AB085 busulfan Unrelated donor hematopoietic stem cell transplantation I. Esteves1,*, J. Fernandes1, A. Ribeiro1, F. Santos1, G. Perini1, (UD-HSCT) in acute myeloid leukemia (AML) using a R. Helman1, C. Castro Ju´nior1, V. Mattos1, A. Kondo1, A. Seber2, reduced intensity conditioning (RIC) regimen and J. S. Oliveira3, J. C. Barros4, V. Colturato5, N. Hamerschlak1, thymoglobulin (TMG) dose according to HLA disparity F. Kerbauy1 E. L. D. S. Espada1, A. Alho1, P. Santos Sousa1, C. Martins1, 1Hematology and Stem Cell Transplantation, Hospital Israelita F. Lourenc¸o1, R. Moreno1, H. Martins1, A. R. Ferreira1, M. Neves1, Albert Einstein, 2Grupo de Apoio ao Adolescente e a` Crianc¸a com I. Conde1, S. Mendes1, D. Alves1, A. Rodrigues2, C. Juncal2, Caˆncer - GRAAC, 3Disciplina de Hematologia e Hemoterapia - N. Camacho1, J. Forjaz de Lacerda1,*, J. Alves do Carmo1 Universidade Federal de Sa˜o Paulo, 4Hospital Brigadeiro, Sao 1Servic¸o de Hematologia e Transplantac¸a˜o de Medula, Paulo, 5Hospital Amaral Carvalho, Jau´, Brazil 2Servic¸o de Imunohemoterapia, Centro Hospitalar Lisboa Norte, EPE - Hospital de Santa Maria, Lisboa, Portugal Introduction: Busulfan (Bu) is used in the conditioning regimen for hematopoietic stem cell transplantation (HSCT). Introduction: HSCT is standard therapy in the treatment of Therapeutic drug monitoring (TDM) of Bu with adjustments higher risk and relapsed AML. In this study we hypothesized doses based on a ‘‘target’’ therapeutic concentration may that the addition of 2 mg/kg of TMG in patients with at least reduce toxicity after HSCT. one HLA allele mismatch is effective in the prevention of the Materials (or patients) and methods: From january 2009 to added risk of severe graft-versus-host disease (GvHD) and january 2014 we prospectively analyzed 42 patients with mortality after UD-HSCT using a RIC regimen. Primary diagnosis of acute myeloid leukemia (AML) and acute lympho- objectives: engraftment, grades II-IV acute GvHD (aGvHD) blastic leukemia (ALL) who underwent TDM of Bu (IV or oral) and moderate to severe chronic GvHD (cGvHD) in donor- before transplantation (test dose) and TDM on 1st day of receptor pairs treated with different doses of TMG according conditioning regimen. At the same time we analyzed 21 patients to HLA matching. Secondary objectives: overall survival (OS), in the retrospective group who did not underwent TDM (from disease-free survival (DFS), treatment-related mortality at day 2004 to 2010). We excluded autologous HSCT, Cord blood HSCT þ 100 (TRM), non-relapse mortality (NRM) and relapse. and haploidentical matched related donor (MRD) transplantation. Materials (or patients) and methods: All patients with AML Results: In the retrospective group (n ¼ 21), all of them submitted to UD-HSCT received a RIC regimen using underwent to MRD transplantation. Six were in first complete fludarabine 150 mg/m2, melphalan 140 mg/m2, TMG e pre- remission (CR1), 18 patients received Bu and cyclo- dnisone, with the possibility of added cytarabine (Ara-C). In phosphamide (BuCy). The median of CD34 þ cells was two consecutive periods from May.’07 to May.’14 the dose of 5.4 106/kg. The second group consisted by patients that TMG used was 8-10 mg/kg (G1) and 4-6 mg/kg; lower doses in received oral Bu (n ¼ 21): 7 underwent to matched unrelated donor-receptor pairs with 10/10 HLA-allele matching and donor (MUD) transplantation and 14 to MRD transplantation. higher doses in pairs with at least one mismatched HLA-A/-B/- Eight patients were in second complete remission (CR2), 4 had Cw/-DRB1 allele. Post-transplant GvHD prophylaxis: cyclos- active disease status and the remainder had unknown "status". porin A and mycophenolate mofetil. Data analysis: SPSSs v22. Fifteen received BuCy and 16 received cells from PBSC. The Results: n ¼ 34 (M ¼ 35.3%; F ¼ 64.7%); median age 50.1 years median of CD34 þ cells was 5.8 106/kg. The median area (18-67). 58.8% of patients transplanted in first complete under the curve (AUC) in 24 hours was 4950 mMol.min (3196.6- remission (CR1), 26.5% in CR2 and 14.7% in partial remission 8212 mMol.min). The third group was IV Bu (n ¼ 21): 7 patients or with advanced disease (PR/AD). There were less patients underwent MRD and 14 MUD transplantation, 7 patients were transplanted in CR1 in the lower TMG dose group (G2), albeit CR1, 7 had active disease or prior HSCT with a mean of age not significantly (70.0% vs. 42.9%; Fisher p-value: 0.163). Ara-C 52.7 years (20-74). The predominant conditioning was was used in all patients of G1, and in G2 only in most of those

S590 with PR/AD (100.0% vs. 35.7%; Fisher p-value: o0.001). HLA persisted. He consecutively developed a severe GvHD-IV (skin, matching was identical between groups: 10/10 ¼ 26.5%, 9/ liver, gut) and died. In the Thio-group 1 patient presented with 10 ¼ 64.7%, 8/10 ¼ 8.8%. Engraftment rate was 100%. HLA an early relapse of his AML. The median leukocyte engraft- mismatch was not correlated with acute GvHD, chronic GvHD ment was in all patient on day þ 18 (range 11-60), in the Thio- or death both within and between groups (total aGvHD 51.5% group on day þ 16 (range 12-60) and in the Mel-group on day and cGvHD 20%). Patients with mismatches had a tendency þ 24 (range 11-34). The median time of neutrophile and for lower relapse (4.2% vs. 33.3%; Breslow p-value 0.052). The platelet recovery was on day þ 19 and day þ 20, respectively source of the graft was not correlated with GvHD. (Thio-group day þ 17 and þ 16; Mel-group day þ 28 and At a median follow-up of 542 days (67-2768): OS ¼ 58.8%, þ 31). After a median follow up of 16,5 months (range 1-44) DFS ¼ 52.9%, Relapse ¼ 12.1%. TRM ¼ 8.8% and NRM ¼ 37.9%. only 1 patients developed a severe stage IV GvHD (gut, liver, G2 had better OS (45.0% vs. 78.6%; Breslow p-value: 0.048), skin) and 1 patient (Mel-group) with stage III GvHD of the gut with a trend towards better NRM (50.0% vs. 18.2%; Breslow which responded very well to steroids and ECP. Nine patients p-value: 0.061). Relapse was greater, but not significantly, in G2 (6 Thio-group, 3 Mel-group) developed Stage II GvHD of the (5.3% vs. 21.4%; Breslow p-value: 0.158), and DFS was not skin which all responded very well to steroids. Four patients (2 significantly different in the two groups (45.0% vs. 64.3%; in each group) developed severe VOD which responded to Breslow p-value: 0.169). TRM did not differ significantly (15.0% defibrotide treatment. vs. 0.0%; Breslow p-value: 0.137). Conclusion: In this preliminary retrospective study, Treosulfan In patients transplanted without Ara-C in the conditioning and Fludarabine based conditioning regimen is a promising regimen there was greater OS (88.9% vs. 48.0%; Breslow regimen in patients with malignant or non-malignant diseases p-value 0.067) and greater relapse (33.3% vs 4.2%; Breslow undergoing HSCT. There were no significant differences in p-value 0.021), with no difference in DFS. Treo/Flu based conditioning regimens combined with Thio Conclusion: WiththeRICregimenusedwewereableto or Mel. The use of Treo/Flu based conditioning suggests overcome the deleterious effect of HLA mismatch in GvHD acceptable outcomes in children undergoing HSCT for a wide incidence and mortality, leading to good DFS with a low relapse range of diseases. rate. With the TMG dose reduction used we were able to further Disclosure of Interest: None declared. increase OS and reduce NRM. It must be noted that patients transplanted in G2 tended to havemoreadvanceddiseaseand AB087 did not receive Ara-C as part of their conditioning regimen. Abstract Withdrawn Disclosure of Interest: None declared.

AB086 Treosulfan and Fludarabine based Conditioning AB088 regimen in Pediatric Patients undergoing Allogeneic Hyper-fractionated TBI exerts a stronger anti-leukemic Hematopoietic Stem Cell Transplantation: A single effect than single dose TBI in HLA matched related T-cell centre experience depleted hematopoietic stem cell transplantation for 1,* 2 1 1 1 adults with acute leukemia J. Foell , K. Buerle , A.-K. Dannenmaier , M. Jakob , K. Rehe , 1,* 1 1 1 1 S. Corbacioglu1 L. Amico , C. Aristei , L. Ruggeri , A. Terenzi , F. Falzetti , 1 2 L. De Carolis1, F. Aversa2, M. F. Martelli1, A. Velardi1, A. Carotti1 Pediatric Hematology/Oncology, KLINIK UND POLIKLINIK FUER 1 2 KINDER- UND JUGENDMEDIZIN, Regensburg, Germany University of Perugia, Perugia, University of Parma, Parma, Italy

Introduction: The use of the alkylating agent Treosulfan (TF), a Introduction: Over the past two decades several studies structural analog of busulfan, in hematopoietic stem cell demonstrated that, for adults with acute leukemia,T-cell transplantation (HSCT) conditioning regimens demonstrated depleted hematopoietic stem cell transplantation from HLA already acceptable post-transplant outcomes in children and matched sibling donors and a TBI-based myeloablative in adult patients. TF combined with fludarabine (FL) has been conditioning regimen were associated with low incidences shown to be a promising combination in terms of myeloa- of acute graft versus host disease (aGvHD) and transplant blativion, immunosuppression and anti-leukemic efficacy related mortality (TRM) as well as a graft versus leukemia effect associated with as limited organ toxicity in patients with that was as good as in unmanipulated graft transplants. non-malignant or malignant diseases. Despite these achievements, the most significant cause of Materials (or patients) and methods: A retrospective analysis transplant failure is still leukemia relapse. To determine of 24 patients (15 male, 9 female, median age 10,7 years) with whether single fraction versus hyper-fractionated TBI sche- malignant disease (5 ALL, 5 AML, 1 CML, 1 MDS, 1 HLH, 1 dules impacted upon leukemia relapse, we retrospectively Neuroblastoma) and non-malignant disease (6 SCD, 1 IL-10 analyzed outcomes of 35 T-cell depleted matched transplants receptor defect, 1 IPEX-like syndrome, 1 ALPS, 1 M. Crohn) we performed at our centre. underwent allogeneic HSCT between April 2011 and Decem- Materials (or patients) and methods: From December 2005 to ber 2014. Thirty-three from matched unrelated donor (MUD), 7 October 2013, 35 patients with acute leukemia (26 AML, 9 ALL, 32 from haploidentical donor (MMRD, CD3/CD19 depleted grafts) in CR, 3 in relapse, median age 47 years), were transplanted from and 4 from matched related donor (MD). Preparative regimen HLA matched siblings in our centre. The conditioning regimen included ATG (3 x 20 mg/kg), Fludarabine (4 x 40 mg/m2), for 19 patients included 8 Gy single TBI (sTBI, at a median dose- Treosulfan (3 x 14 g/kg) and Thiotepa (2 x 5 mg/ kg, 17 rate of 10.7 cGy/min, lung dose 4 Gy) while 16 patients received patients) or Melphalan (2 x 70 mg/m2, 7 patients). GvHD hyper-fractionated TBI (HFTBI, 1.2 Gy 3 times a day for 4 days up to a total dose of 14.4 Gy, lung dose 9 Gy). Both groups also prophylaxis in malignant diseases consisted of CsA and MTX 2 and in non-malignant diseases CsA and MMF. received Thiotepa (10 mg/kg) and Fludarabine (160 mg/m ). No Results: The enrolled 24 patients were subdivided in 2 groups post-transplant pharmacological immunosuppression was given. according to conditioning TF/FL with Thiotepa (Thio) or Prognostic factors were equally distributed in the two TBI groups. Melphalan (Mel). The chemotherapy was well tolerated in all The Kaplan-Meier method evaluated leukemia-free survival (LFS). patients. The overall survival (OS) and disease free survival Cumulative incidence (CI) estimates were used for relapse and (DFS) was 84% (20/24) and 3 patients died in the Thio-group TRM, as they are competing risks. Multivariate analysis assessed (ALL, AML, HLH) and 1 patient in the Mel-group (AML) because the impact of diverse variables on transplantation outcomes. All P of disease recurrence. We achieved a successful engraftment values were two-sided and were considered significant at Po.05. in 23 patients, but 1 patient in the Thio-group, with HLH and Results: Engraftment was successful in 34/35 evaluable second SCT, showed a delayed engraftment around day þ 60 patients. The CI of aGvHD (grade 42) was 14%. No patient post-SCT. Despite engraftment the hemophagocytosis developed chronic GvHD. The CI of TRM was 14%: 10% in

S591 HFTBI group vs 20% in sTBI group (P ¼ n.s.).The CI of leukemia leukemia patients. Further prospective studies with larger relapse was 25% (9/35 patients). It was 0% in HFTBI group and series are required on this issue. 45% in sTBI group (P ¼ 0.04). At a median follow up of 4 years, Disclosure of Interest: None declared. 21 patients are alive and leukemia free survival is 65% (80% in HFTBI group vs 45% in sTBI group, P ¼ 0.03). This significant difference was confirmed by multivariate analysis which AB090 included variables such as disease, disease status, etc. Efficacy of CEAM regimen in hematopoietic stem cell Conclusion: In demonstrating that hyper-fractioned TBI transplantation for relapsed or refractory Hodgkine exerted a stronger anti-leukemic effect than single dose TBI, lymphoma our results appear in line with findings from in vitro and M. Mehdizadeh1,*, A. Hajifathali1 murine models of radiobiology studies which showed that 1Taleghani BMT center, Shahid Beheshti university of medical fractionated TBI schemes exerted greater myeloablative sciences, Tehran, Iran, Islamic Republic Of efficacy. The present study confirmed that, for adults with acute leukemia, T-cell depleted hematopoietic stem cell Introduction: Autologous stem cell transplantation (ASCT) is transplantation from HLA matched sibling donors and TBI- the optimal treatment strategy for Hodgkin lymphoma (HL) based myeloablative conditioning, prevented GvHD and patients unresponsive to first course of therapy or relapsing provided a good LFS rate with low TRM. In this transplant after primary treatment. We analyzed our registry data for setting, HFTBI was associated with reduced risk of relapse, no patients with Hodgkine lymphoma in Taleghani bone marrow evidence of increased toxicity and consequently better center affiliated to Shahid Beheshti university of medical outcomes than single dose TBI. Given our small cohort of sciences patients, data from a larger series are needed to confirm Materials (or patients) and methods: All the patients with present observations. refractory or relapsed HL who underwent a first autologous or Disclosure of Interest: None declared. a first or second allogeneic HSCT between 2007 and 2013 were included.The conditioning regimen for autologus transplanta- AB089 tion consisted of CCNU 200 mg/square meter,Cytozar Comparison of the conditioning regimens with or without 1200 mg/ square meter.Etoposide 300 mg/ square meter and TBI in acute lymphoblastic leukemia patients: Experience Melphalan 140 mg/ square meter.We also performed allogenic of a single center reduced intensity transplantation in patient who had post 1,1,* 1 1 1 1 autologous HSCT relapse.Patients were followed for a mean of M. Yeral , C. Boga , H. Ozdogu , S. Solmaz , A. P. Korur , 3 years. S. Asma1, I. Kozanoglu1 1 Results: Autologous and allogeneic HSCT were performed in Bone Marrow transplant Center, Baskent University, Adana, 96 and 6 patients, respectively. Three years Overall survival Turkey (3yOS) and event free survival was 83% and 67% respectively. We performed allogenic reduced intensity transplantation in 6 Introduction: Allogeneic hematopoietic stem cell transplanta- refractory or post autologous HSCT relapse. Out of them 3 is in tion (PSCT) is one of the standard treatment approaches complete remission or stable disease. Grade 2 acute GVHD particularly in high risk patients with a proper performance. occurred in one patient who relapsed later and chronic GVHD Total body irradiation (TBI) –based conditioning regimens are was also occurred in one patient who is in CR.16 patient conventional and widely used. However there are limited passed away,11 out of them were refractory to autologous numbers of studies comparing TBI regimens with other HSCT. Disease status at transplantation,disease stage at last busulphan-containing regimens. relapse, the number of previous chemotherapy courses and Materials (or patients) and methods: Allogeneic PSCT was age were prognostic factors. OS was favorable even in patients applied to a total of 36 acute lymphoblastic leukemia patients who underwent autologous HSCT in disease status other than in Adult Bone Marrow Transplantation Unit of Adana Hospital, complete remission.A first allogeneic HSCT without a previous Baskent University between April 2004 and November 2013. autologous HSCT was performed only in one patient who Of the patients, 19 were female and 17 were male with median relapsed early even after transplantation. age of 30 (range 17-50). 27 (75%) patients had B cell ALL and Conclusion: CEAM regimen as conditioning in autologous 9(25%) had T cell ALL and BCR/ABL was positive in 31% of the HSCT is effective and even curative in patients with relapsed patients. All patients received hyper-CVAD treatment prior to and refractory HL It is preferred to performed as soon as PSCT. FLAG was used as salvage therapy in case of refractory possible in patients who responded to salvage chemotherapy. or relapsed disease. Of the patients, 88% were in complete Allogeneic HSCT is feasible with little complications and might remission when starting PSCT regimen. Transplantation was be beneficial in patients who relapsed after autologous HSCT. applied from HLA matched siblings in all patients. Disclosure of Interest: None declared. Results: Allogeneic PSCT conditioning regimen included Busulphan (16 mg/kg), Fludarabine (150 mg/m2), and ATG- fresenius (30 mg/kg) in 10 cases, TBI (12-13,2GY) and Cyclopho- AB091 sphamide (120 mg/kg) in 26 cases. Cyclosporine and short BUSULFAN-CYCLOPHOSPHAMIDE-ETOPOSIDE (BuCyE) duration of methotrexate were used for GvHD prophylaxis. HIGH-DOSE CHEMOTHERAPY FOLLOWED BY Neutrophil and platelet engraftment was observed on median AUTOLOGOUS STEM CELL TRANSPLATATION IN 12 day (10-15) and 11 day (8-18), respectively. Patients were LYMPHOMA PATIENTS: RESULTS OF A SINGLE administered trimethoprim-sulphometoxazole, fluconazole and INSTITUTION acyclovir prophylaxis. Median disease-free survival was 12 M. Sonmez1,*, H. M. Ozbas1, N. Ermantas1, S. Yuzbasıoglu1, months, overall survival was 17 months (1-56). Mortality rate C. Sehit 2 was found as 4/36 (11%) within the first 100 days. One year 1Karadeniz Technical University Medical Faculty, Haematology survival was 47% and 3 year survival was 16%. Overall survival Division, 2Karadeniz Technical University Medical Faculty, was median 13 (2-51) and 19 (1-56) months for Busulphan- Department of Internal Medicine, Trabzon, Turkey fludarabine-ATG and TBI-cyclophosphamide conditioning regi- mens, the difference was statistically significant (Po0,05). Cox Introduction: Autologous haematopoietic stem cell transplan- regression analyses revealed that BCR/ABL and disease condi- tation (AHSCT) seems to be the treatment of choice in patients tion at the time of PSCT were effective on disease-free survival younger than 65 years who failed after first line therapy. In and overall survival. preparation of AHSCT, several regimes were on trial with no Conclusion: This retrospective study indicates that TBI and clear conclusion. In this study we evaluated the results of a cyclophosphamide-based conditioning regimens are superior regime (BuCyE, busulfan-cyclophosphamide-etoposide) in to other conditioning regimens in acute lymphoblastic preparation of AHSCT.

S592 Materials (or patients) and methods: Eighteen patients (12 graded and the length of time each was reported was male, 6 female; mean age 39; age range: 17-61) were included recorded. Data for 20 patients on each regimen was gathered. into the study (period: January 2010 to June 2014). Nine of the Results: The results were compared using a Mann-Whitney patients had Hodgkin Lymphoma (HL; 5 nodular sclerosing, 4 U test, and are as follows: mixed cellular), 5 of the patients had diffuse large B-cell Mucositis, diarrhoea, neutrophil recovery and length of time lymphoma (DLBCL), 2 of the patients had mantle cell on TPN showed no significant difference (P4 ¼ 0.05) between lymphoma (MCL), 1 patient had peripheral T-cell lymphoma, the regimes. A significant difference for platelet recovery 1 patient had anaplastic large cell lymphoma. Four patients was shown at P4 ¼ 0.05, platelet recovery with LEAM was (2 with HL and 2 with MHL) had bone marrow infiltration. longer. However, BEAM had several outlying figures therefore Seven of HL patients had complete remission, while the other the group may not reflect the statistical data. LEAM resulted 2 had partial response; 5 of non-Hodgkin Lymphoma had in slightly worse nausea and vomiting related toxicities: 2 complete remission, while 1 MCL patient, 2 patients with patients were classed as having grade III toxicity compared to DLBCL (1 with primary CNS-DLBCL) and 1 patient with those observed in BEAM who achieved toxic grade of I-II anaplastic large cell lymphoma had partial response; Following (n ¼ 3, n ¼ 16 respectively). the last chemotherapy administration, induction with G-CSF Conclusion: In conclusion LEAM is a suitable replace- were done and the stem cells of the patients were collected. ment for BEAM as it has similar toxicity and count Busulfan (days -7, -6, -5 and 3.2 mg/kg/day), etoposide (days -5, recovery. This study supported Perfetti at al findings. -4 and 400 mg/m2/day), cyclophosphamide (days -3, -2 and A follow on from this study would be to repeat the study in 50 mg/kg/day) and MESNA (days -3, -2, -1) were given in a year time and review the patients’ disease status for the preparation of AHSCT. different regimens. Results: Mean time to engraftment was 10 days (9-11 days). References: Reference Perfetti, P., Scollo, C., Terruzzi, E., Parma, No life-threatening complication or intolerance to BuCyE was M., Pogliana, E. and Pioltelli, P. (2007) Modified BEAM schedule noted. All HL patients had complete response following (lomustine instead of carmustne) in the conditioning of ABMT AHSCT, while two of them also needed XRT for the primary for lymphoid malignancies. Bone Marrow Transplantation 43 bulky mass. Four patients having non-Hodgkin Lymphoma (Sup 1): S206-S207 with partial response to primary treatment were lost 4 to 6 Disclosure of Interest: None declared. months after AHSCT. A patient having peripheral T-cell lymphoma who had disease progression was given anti- AB093 CD30 and survival lasting for 16 months was noted. Remaining Follow-up Data of Fludarabin-Cytarabine-Melphalan 3 patients with DLBCL and 1 patient with MCL were in Combination as high dose chemotherapy conditioning remission. regimen for autologous peripheral blood stem cell Conclusion: Our preliminary results suggested that BuCyE transplant in Hematology malignancies – Hospital may be an effective alternative in preparation of AHSCT. Ampang experince References: 1-Kim JE, Lee DH, Yoo C, Kim S, Kim SW, T. C. Ong1,*, M. Z. Zakaria1, Y. K. Guan2, J. S. Rajasuriar1, J. Tan1, Lee JS, Park CJ, Huh J, Suh C. BEAM or BuCyE high-dose 1 1 chemotherapy followed by autologous stem cell trans- S. M. Tan , K. M. Chang 1Haematology, HOSPITAL AMPANG, MALAYSIA, Pandan Mewah, plantation in non-Hodgkin’s lymphoma patients: a single 2 center comparative analysis of efficacy and toxicity. Leuk Res. Internal Medicine, Hospital Melaka, Melaka, Malaysia 2011;35:183-187. 2-Ayala E, Tomblyn M. Hematopoietic cell transplantation for Introduction: Fludarabine-Cytabarine-Melphalan combina- lymphomas. Cancer Control. 2011;18:246-257. tion(FLAM) is one of conditioning regimens used in our center 3- Yoon DH, Lee DH, Choi DR, Sohn BS, Kim S, Kim SW, Lee JS, which theoretically has the advantage of being able to provide Lee SW, Huh J, Suh C. Feasibility of BU, CY and etoposide Central Nervous System penetration (high dose cytarabine) as (BUCYE), and auto-SCT in patients with newly diagnosed well as 2 non-cross resistant chemotherapy drugs (Fludarabine primary CNS lymphoma: a single-center experience. Bone and Melphalan) that many patients were previously not Marrow Transplant. 2011;46:105-109. exposed to. We report the updated data on bigger and longer Disclosure of Interest: None declared. follow-up of our cohort conditioned with this regimen (previously presented as poster in EBMT 2011 and 2013). Materials (or patients) and methods: Selected patients AB092 with prior autologous peripheral blood stem cells Comparative toxixity study of LEAM versus BEAM collected (chemotherapy þ GCSF of GCSF mobilisation) were chemotherapy in lymphoma patients undergoing treated in Hospital Ampang from June 2011 till May 20414 autologous pheripheral blood stem cell transplants with IV Fludarabine 30 mg/m^2 for 5 days, IV Cytarabine S. Potts1,* 2000 mg/m^2 for 5 days, IV Melphalan 140 mg/m^2 for 1WESTERN GENERAL HOSPITAL, Edinburgh, United Kingdom 1 day before autologous stem cell reinfusion. Most of the patient received GCSF when neutropenic until engraftment. Introduction: BEAM (carmustine, etoposide, cytarabine Demographic data, disease and remission status and and melphalan), is a common transplant regimen chemotherapy toxicity, engraftment details and follow-up used in lymphoma patients but due to stock shortages of data were collected. carmustine a search was undertaken for an alternative Results: There were a total of 55 patients treated. 45(81.8%) regimen that would provide similar count recovery and had Non-Hodgkin’s Lymphoma in which 35(63.6%) were toxicity profile. B cell lymphoma and 10(18.2%) were T cell lymphoma. The search discovered a report by Perfetti at al (2007) that 9(16.4%) had Hodgkin Lymphoma and 1(1.8%) had looked at substituting lomustine for carmustine: LEAM relapsed Acute Promyelocytic Leukemia with CNS (lomustine, etoposide, cytarabine and melphalan). However involvement. Median age at transplant of the cohort is 47 Perfetti at al, had not compared LEAM to BEAM. Therefore this (16-67). In term of mucositis, 53.7% had no significant oral study aimed to compare the different regimes in terms of mucositis, 27.8% had grade 1, 11.1% grade 2, 7.4% toxicities, as graded by the NCI Common Toxicity Criteria. grade 3 and no grade 4 mucositis noted. There were no Materials (or patients) and methods: Retrospective data was grade 3 or grade 4 renal or liver toxicities. Median autologous gathered on the following toxicities: mucositis, diarrhoea, peripheral stem cell dose was 3.89 106 CD34/kg (1.97–49.24). nausea/vomiting, TPN use, neutropenic fever, death, time for Median day of neutrophil engraftment was D þ 10 (8–15) and neutrophil counts to recover to median day of platelet engraftment was D þ 11(8–17). All 40.5 109/l and time for platelet count to recover to patients except one with very early mortality engrafted. There 420 103 (without transfusion support). The toxicities were was 1 early treatment related mortality (TRM)(1.8%) due to

S593 sepsis in a patient with advanced refractory Hodgkin AB095 Lymphoma, and 3(5.5%) late TRM due to infections (2 from Chemosensitization of acute myeloid leukaemia cell line reactivation with fulminant Hepatitis B after defaulted antiviral, following mobilization by AMD3100 and 1 from presumed infection with unidentified source). Z. Shen1,*, P. kong1, D. zeng1, X. zhang1,Y.ma1 Median follow-up from transplant was 29 months (7-41). 15 1Department of Hematology, Xinqiao Hospital, the Third Military other died from relapsed and progressive disease,7 of them Medical University, chongqing, China were refractory when transplanted. Estimated overall survival at 2 years is 62%. Introduction: It is recognized as the refractory AML. While the Conclusion: FLAM is a feasible and effective high dose abnormal hematopoietic stem cell niche with high adhesion chemotherapy regime that could be used in combination with and proliferation may be the root cause of the resistance and autologous stem cell rescue. Early TRM was acceptable and relapse[1].Stromal cell derived factor-1 (SDF-1), excreted by complications appeared manageable, reactivation of Hepatitis bone marrow osteoblasts, might be involved in shielding of B needs special attention. Engraftment rates were comparable bone marrow environment from leukaemia cells by binding to with other regimes; outcome appears to be not inferior in its receptor CXCR4, but the relationship between SDF-1/CXCR4 good risk patients. Long term follow-up of this cohort will axis and leukaemia cells is still a mystery. reveal more information. Materials (or patients) and methods: The experiments were Disclosure of Interest: None declared. built on the 3D PS scaffolds coated with osteoblasts. First we seeded bone marrow stromal cells (MSCs) on the scaffolds and AB094 induced them into osteoblasts and then leukemia cells were Preliminary results of Jurkat cell-reactive anti-T seeded on the scaffolds. AMD3100 and cytrarabine were lymphocyte globulin for stem cell transplantation added to observe the adhesion rate, migration state, apoptosis in pediatric severe aplastic anemia rate, and cell cycle of leukaemia cells. T. Yang1,*, J. H. Ren1, X. Yuan1, Z. Chen1,J.Hu1 on behalf of Results: Osteoblasts protect leukaemia cells from spontaneous Union group and Ara-C-induced apoptosis (Po0.05) and leukaemia cells 1Department of Hematology, Union Hospital,Fujian Medical were kept in the G0/G1 phase (Po0.05). AMD3100 inhibited University, Fuzhou, China the role of SDF-1/CXCR4 and disrupted the adhesion and migration between leukemia cells and the niche, and Introduction: To assess the effects of Jurkat cell-reactive anti- enhanced the killing effects of cytrarabine T lymphocyte globulin (ATG-F) in the setting of hematopoietic Conclusion: AMD3100 could be as an adjunct to chemother- stem cell transplantation (HSCT) in pediatric severe aplastic apy in patients with refractory AML, as a strategy to sensitize anemia (AA). leukaemia cells to chemotherapy and improve clinical Materials (or patients) and methods: We retrospectively outcomes. reviewed our HSCT experience using ATG-F in pediatric severe References: 1.Amanda Parmar, Stefanie Marz, Sally Rushton, aplastic anemia. Of the total 10 patients included in the study, et al. Stromal Niche Cells Protect Early Leukemic FLT3-ITD þ 4 were male and 6 female with a median age of 9 years (2- Progenitor Cells against First- Generation FLT3 Tyrosine Kinase 14y), including 1 case of EB virus-associated AA, 3 cases of Inhibitors. Cancer Res, 2011,71 (13):4696-4706. hepatitis-associated AA (HAA) and 1 case of pure red cell Disclosure of Interest: None declared. aplasia. Before HSCT, 5 of 10 patients received long-term oral supplement with cyclosporin A (CSA), androgen, or traditional Chinese medicines, and 2 were unresponsived to one course of rabbit antithymocyte globulin ATG. The median time to HSCT is 15.5 m (3-60 m). Of them, 6 patients were trans- Minimal residual disease, tolerance, planted from 10/10 HLA matched related donors, 1 was from chimerism and immune reconstitution syngeneic donor, 1 was from 7/10 related donor, 1 was from 10/10 unrelated donor and 1 was from 9/10 unrelated donor. The conditioning includes cyclophosphamide, fludarabine and ATG-F (10 mg/kg for three days). All AB096 transplant recipients received peripheral blood as the source Meaning of mixed chimerism by short tandem repeat of stem cells, except 3 had co-infusion with bone marrow- PCR in allogeneic stem cell transplantation derived stem cells. The GVHD prophylaxis consisted of B. Kim1,*, J.-L. Choi1, K.-S. Woo1, K.-H. Kim1, J.-Y. Han1 ATG, CsA, MTX and anti-CD25. 8 of 10 patients experienced 1Department of Laboratory Medicine, Dong-A university medical severe infections before HSCT, and 3 have liver function failure center, Busan, Korea, Republic Of with HAA. Results: All patients achieved successful engraftment with Introduction: Monitoring of chimerism of hematopoietic cells complete donor chimerism. The most striking results were by conventional short tandem repeat (STR)-PCR is the standard that neutrophil engraftment was obtained on day10 (8-15d), of follow patient with hematologic disease after allogeneic and thrombocyte on day15 (11-18d). 7 of 10 patients (70%) stem cell transplantation. Mixed chimerism is clinically remain alive without graft failure after a median of 12 months important poorly defined aspect of diseases. Accurate (range, 2–24 m). Liver function improved during the trans- determination of chimerism status is important, we evaluated plantation procedure in 3 patients with previous liver the meaning of mixed chimerism determined by STR in function failure. Sepsis were the main complications in 5 of various hematologic diseases. 10 patients, including 1 with lethal multi-drug resistant Materials (or patients) and methods: 360 samples from 64 sepsis. Another 2 patients died of severe extensive aGVHD recipient who underwent allogeneic HSCT at Dong-A medical or cerebral hemorrhage even with the thrombocyte recovery, center, Busan, Korea between 2011and 2014 were included in respectively. In a multivariate analysis, prior ATG treatment this study. The recipients consisted with acute myeloid and the interval of more than 6 months from diagnosis to leukemia (n ¼ 28), acute lymphoblastic leukemia (n ¼ 15), transplantation were independent adverse risk factors for aplastic anemia (n ¼ 9), myelodysplastic syndrome (n ¼ 5), survival. myeloproliferative neoplasm (n ¼ 4), Paroxysmal nocturnal Conclusion: The conditioning with ATG-F appears excellent hemoglobinuria (n ¼ 2), lymphoma (n ¼ 1), Wiskott–Aldrich outcome with expediting hematopoietic recovery. Earlier syndrome ( n ¼ 1). Sixteen STR loci (D8S1179, D21S11, D7S820, transplantation after diagnosis without immunosuppressive CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, treatment can achieve better outcomes. D19S43, vWA, TPOX, D18S51, Amelogenin, D5S818, FGA) were Disclosure of Interest: None declared. used to evaluate chimerism monitoring. The results were

S594 compared with bone marrow aspiration examination, cytoge- death or relapse before the second time-point for analysis or netic study, FISH and clinical manifestation lack of financial resources for further determination of Results: The mixed chimerism was observed in 9 of 28 chimerism. Lineage-specific chimerism (LSC) was performed (32.14%) patients with AML, 8 of 15 (53.33%) ALL, 4 of in 21 patients (52.5%). Based on the dynamics of LSC, in 4 9(44.44%) AA, 3 of 5 (60.00%) MDS, 1 of 4 (25.00%) CML, 1 of 2 patients presenting PMC, especially affecting the T-cell (50.00%) PNH. By the engraft type, mixed chimerism was compartment, withdrawal of immunosuppressive treatment observed in 2 of 7(28.57%) patients with BMT, 24 of resulted in SMC in two of them, while in the other 2 patients 59(40.68%) with PBSCT. During follow up, the rate of the dynamic of MC continued to be progressive. 25% of the discrepancy was 18.33% (66/360) between STR and other patients presented grade I/II acute GvHD. Of them, 40% with analyzed results or clinical manifestatio. Of 66 discrepancy SMC and the rest with CDC. Among the patients with samples, 7 (7/79, 8.86%) was from patients with BMT and malignant diseases, 27% relapsed, out of them 43% having 59(59/256, 23.05%) was from PBSCT. 47 samples of 360 CDC at the last chimerism assessment prior to relapse. (13.06%) represented mixed chimerism but the results of other Probability of overall survival at 1 year post-transplant was study was normal and favorable manifestations. And 19 85.33% for patients with CDC and 75% for those with MC samples of 360 (5.27%) represented complete donor chimer- (P ¼ 0.045, CI:95%). ism but their morphological or cytogenetic results revealed Conclusion: In our experience survival of patients at 1 year relapsed disease with poor clinical manifestation. When we post-transplant was significantly superior in patients with analyzed the latest outcome, favorable prognosis with mixed CDC as compared with those with MC. Prompt withdrawal chimerism were detected in 2 of 3 AA(66.67%) and 1 of 9 of immunosuppression proved to be efficient in half AML(11.11%). of the patients with PMC. The proportion of patients with Conclusion: In this study, mixed chimerism was not always acute GvHD was similar in patients with CDC/MC. Lineage- related with poor prognosis, favorable prognosis was observed specific chimerism analysis during the post-transplant more frequent in AA and AML recipients. Reconstitution of period is a predictive tool for the outcome of patients with normal hematopoietic elements are thought to has caused the alloHSCT. mixed chimerism. So we suggest mixed chimerism by STR Disclosure of Interest: None declared. should be interpreted carefully with other studies and more specific and more sensitive method is necessary for identifying AB098 disease relapse or poor condition. Monitoring of haematopoietic chimerism after allogenic Disclosure of Interest: None declared. stem cell transplantation: a single paediatric centre experience AB097 D. Di Martino1,*, M. Di Duca2, M. P. Terranova3, S. Giardino3, Prediction of outcome by chimerism analysis in allogeneic M. Faraci3, G. Morreale3, E. Lanino3 hematopoietic stem cell transplantation 1Stem Cell Laboratory-Haematology and Oncology Department, C. Jinca1,*, S. Arghirescu2, A. M. Balan1, A. Oprisoni1, A. Isac3, 2Laboratory of Pathophysiology of Uremia, 3SCT Unit- Haema- A. Pascalau4, C. Popa5, E. Gai6, M. Serban1 tology and Oncology Department, IRCCS Pediatric Institute G. 1Bone Marrow Transplantation, University of Medicine and Gaslini, Genova, Italy Pharmacy ‘‘V. Babes’’, 2Bone Marrow Transplantation, University of Medicine and Pharmacy "V. Babes", 3Bone Marrow Transplan- Introduction: Allogenic haematopoietic stem cell transplanta- tation, Emergency Children’s Hospital "L. Turcanu", 4Bone Marrow tion is a potentially curative therapy for neoplastic and non- Transplantation, Emergency Children’s Hospital ‘‘L. Turcanu’’, neoplastic haematological diseases. Post-transplant monitor- 5Department of Genetics, 6Transplant Immunology, University of ing of the proportion of donor and recipient haematopoiesis in Medicine and Pharmacy ‘‘V. Babes’’, Timisoara, Romania the patient (chimerism) is an instrumental tool useful in directing further treatment choices. DNA isolated from Introduction: Assessment of hematopoietic chimerism peripheral blood, bone marrow or specific isolated cell became a standard method for monitoring the presence of lineages is used for short tandem repeat analysis (STR)-PCR, lymphohematopoietic cells of donor origin in recipients of an the gold standard method of chimerism monitoring. allogeneic hematopoietic stem cell transplantation (alloHSCT) Materials (or patients) and methods: We performed and lineage-specific chimerism analysis at specific time-points semiquantitative STR-PCR chimerism analysis at different time proved to have predictive values for the outcome in the subset points post transplant on 221 children who underwent of patients with mixed chimerism (MC). haematopoietic stem cell transplantation (HSCT) at G.Gaslini Materials (or patients) and methods: This retrospective Children Hospital and Research Institute over the last 8 years. study, aiming to assess the predictive value of chimerism A measurement of chimerism was performed: at time of analysis on the outcome of patients with alloHSCT, was engraftment and at least monthly in patients with Mixed conducted on a lot of 40 patients who were transplanted in Chimerism until 1 year; at time of engraftment, 30 days, 100 the Centre for Bone Marrow Transplantation Timis¸oara/ days, 6 months until 1 year in patients with Complete Romania between 2003-2014. Chimerism was assessed either Chimerism. Additional cell sorting with Macs columns was by short tandem repeats PCR (STR-PCR) analysis in sex- performed for determination of chimerism in T cells, NK cells, B mismatched donor-recipient pairs or by fluorescence in situ cells and myeloid cells in select children. hybridization (FISH) in sex-matched donor-recipient pairs. 65% Results: Of the 221 patients examined, 106 (48%) showed of the patients were transplanted for malignant diseases Complete Chimerism (CC) at time of engraftment and 115 whereas 35% received transplants for non-malignant diseases. (52%) showed Mixed Chimerism (MC), diagnosed using 82.5% of the patients received myeloablative conditioning PB-alone or combined PB-BM, as summarized in following regimens whereas 17.5% were transplanted with reduced table: intensity conditioning. 90% were related transplants and 10% unrelated. 19.7% of the donor-recipient pairs were mis- matched related or mismatched unrelated transplant Diseases Lymphocytic Haemato- Genetic Metabolic Lymphoma Neuro- Results: At first time point analysis (day þ 30), 30% of the and Myeloid logical Diseases Diseases blastomas patients presented with complete donor chimerism (CDC), the Leukemia non- and Solid rest presenting MC. Stable mixed chimerism (SMC) was neoplastic Tumors established in 15% of the patients, transient mixed chimerism CC 62 28 5 5 3 3 106 (TMC) was detected in 42.5% and progressive mixed chimer- MC 38 43 8 12 8 6 115 ism (PMC) in 17.5%. The rest of the patients had only one 100 71 13 17 11 9 221 chimerism assessment due to one of the following reasons:

S595 Among patients with MC, we could identify: 68/115 (59%) providing useful information and facilitating therapeutic with Transient Mixed Chimerism (TMC); 19/115 (17%) intervention. with Stabled Mixed Chimerism (SMC); 28/115 (24%) with Disclosure of Interest: None declared. Progressive Mixed Chimerism (PMC). When we performed Lineage Specific Chimerism (LSC) analysis, increasing in donor T cells was associated with TMC, whereas decrease in AB099 donor T cells was associated with PMC while a percentage of Post-transplant lymphocytosis impact in patients with donor T cell consistently higher than PB in toto was observed reduced intensity conditioning regimen (RIC) in SMC. O. Lopez Godino1,*,E.Pe´rez-Lo´pez1, M. Cabrero Calvo2, Conclusion: In our experience, the majority of patients S. Alonso2, J. Labrador2,F.Sa´nchez-Guijo2,L.Va´zquez2, transplanted for malignant disease show CC. CC is associated C. del Can˜izo2,L.Lo´pez-Corral1, D. Caballero2 with an increased risk of Graft versus Host Disease (GvHD) 1Hematology department, 2Hospital Universitario de Salamanca, both in non-neoplastic than in neoplastic diseases. TMC is Salamanca, Spain associated with a good outcome and reduces the risk of GvHD, as well as SMC that is observed only in non malignant disease. Introduction: Some patients developed lymphocytosis post- Instead, PMC is associated with increased risk of relapse in transplant, however its role is unknown. We analysed the neoplastic diseases and transplant rejection in non neoplastic incidence of lymphocytosis post-transplant and its potential diseases. Chimerism analysis can increasingly enable early impact in overall survival (OS), graft versus host disease detection of disease relapse and level of engraftment, (GVHD) and infectious complications.

[AB099]

S596 Materials (or patients) and methods: We retro- was observed. Among the cases, 19% (10 of 52) carried spectively analysed 109 patients with Acute Myeloid PRAs, 9 with class I only, 5 with class II only, and 4 Leukemia/Mylodisplasic Syndrome who consecutively with both class I and II specific antibodies. However donor received a RIC allogeneic transplant in our centre between specific antibody (DSA) positivity was not showen in the 1998-2013. patients of PRA positivity. There is no significant value Results: Basal characteristics are showed in table 1. With a between T/B FCXM ( þ ) and PRA positivity. Eighteen median follow-up of 35 months (3-156), the OS at 1 and 5 patients developed an acute GVHD (T/B ( þ ): 5/7) and 2 years was 71% and 60%. The main cause of death was relapse developed a chronic GVHD (T/B ( þ ): 0/2) (P40.05). A in 18% and overall transplant related mortality was 15%. The relationship between Acute or chronic GVHD development median of maximum lymphocytes was 2695/mL(o100-14400); and anti-HLA positivity could not be detected (P40.05). with a median onset of 14,5 months (o1-104). In the Patients with a positive T and B FCXM was no correlation univariate analysis (Kaplan-Meier) significant factors associated between full chimerism status in the 100th day of the after with better OS were chronic GVHD (cGVHD) (yes/no and HSCT (P40.05). grade), stopping inmunosupresive treatment, viral infections, Conclusion: Preformed host antibodies may contribute to achieve complete response (CR) at day þ 100 and lymphocy- graft rejection following hematopoietic stem cell transplanta- tosis 42500 at any time post-transplant (Po0.05); a trend to tion. The association between positive FCXM T / B in HLA better OS was found in late reactivation (those reactivations mismatch graft and rejection were determined in previous after day þ 100) of herpesvirus group and worst if early study. The role of anti-HLA antibody on HSCT are not reactivation (P ¼ 0.08). In multivariate analysis (Cox), all the associated with GVHD, rejection and chimerism in our study. variables except lymphocytosis 42500 maintained their The predictive value of cytotoxic and flow cytometric cross- significance. The association between lymphocytosis 42500 match analysis before allo-HSCT remains still unclear. and development of cGVHD (OR ¼ 5;1.94-12.9;P ¼ 0.01) as well References: 1. Stephen Spellman, Robert Bray, Sandra Rosen- as the relation between stopping of inmunosupresive treat- Bronson et all. The detection of donor directed, HLA-specific ment (OR ¼ 2,47;1,09-5,6; P ¼ 0,03) is responsible of the alloantibodies in recipients of unrelated hematopoietic cell disappearance of lymphocytosis as an independent prognostic transplantation is predictive of graft failure. Blood. 2010; DOI factor in the multivariate analysis. If we focus in infectious 10.1182/blood-2009-09-244525. diseases, lymphopenia o2500 was not associated with fungal 2. Jonathan A Gutman, Susan K McKinney, Shalini Pereira, infections but late reactivation of herpesvirus was associated Sandra L Warnock et al. Prospective monitoring for alloimmu- with lymphocytosis 42500 (OR ¼ 3.5;1.3-9.16;P ¼ 0.01) with or nization in cord blood transplantation: ‘‘virtual crossmatch’’ without active cGVHD. can be used to demonstrate donor-directed antibodies. Conclusion: Although cGVHD and CR at þ 100 were the two Transplantation. 2009; 87(3): 415–418. factors with more impact in OS, there was an association 3. J Mattsson, A Nordlander, M Remberger et al. Cytotoxic between development lymphocytosis 42500 once the crossmatch analysis before allo-SCT is a poor diagnostic tool inmunosupresive treatment has stopped and the develop- for prediction of rejection. Bone Marrow Transplant.2010; 45: ment of cGVHD. Moreover, late herpes viral infections were 235–238. correlated with lymphocytois. Although true implications of Disclosure of Interest: None declared. lymphocyte increase after transplant is not very well known, immune reconstitution for sure plays a role on GVHD and/or AB101 viral infections. More prospective studies should be done to The clinical significance of the monitoring minimal investigate the role of lymphocytosis and if it is any poblation residual disease for maintanence therapy after autologous with specially implication. stem cells transplantation in patients with multiple Disclosure of Interest: None declared. myeloma S. Shamansky1,*, I. Fedotova2, L. Novoseltceva3, C. Melkova1, AB100 O. Pisarevskaya3, O. Rukavitcin4 Importance of Crossmatch and Anti-HLA Antibody 1bmt, Russian Cancer Research Center named NN Blokhin, 2bmt, In Pediatric HSCT Recipients Main Millitary Clinical Hospital, 3hematology, 4hematological S. Usta Akgul1,*, D. Atay2, F. Erbey2, C. Kekik Cınar1, A. Akcay2, center, Main Military Clinical Hospital, Moscow, Russian M. Akbıyık2, F. Savran Oguz1,G.O¨ ztu¨rk2 Federation 1Medicine Faculty, Department of Medical Biyology, Istanbul University, 2Acıbadem University, Atakent Hospital, Istanbul, Introduction: High-dose therapy with autologous stem cell Turkey transplantation (auto-Tx) for treatment of the patients (pts) with multiple myeloma (MM) has been considered the Introduction: Hematopoietic stem cell transplantation (HSCT) standard frontline treatment for younger pts even in the era recipients may become alloimmunized to foreign human of novel agents (bortezomib and lenalidomide). Post- leukocyte antigens (HLAs) through pregnancy or blood transplant treatment has clearly shown improvements in the transfusions. The resulting sensitization may include antibo- depth and duration of response. However, the optimal dies directed against mismatched HLA antigens of a potential posttransplant therapy, if any, is still to be defined, stem cell donor. A lymphocyte cross-match may an effective as well as duration of such treatment. Monitoring of the tool to evaluate alloimmunization and potential donor/ minimal residual disease (MRD) may helps to deside this recipient in compatibility. Prescreening of serum and the problem. identification of specific HLA antibodies could be used as part Materials (or patients) and methods: Since 2001 we of a donor selection strategy designed to avoid a potential performed 85 auto-Tx þ 1 allo-Tx in 74 pts. 71 pts included in deleterious in compatibility. the analysis. 12 pts recieved 2 auto-Tx (6 planned )tandem* n Materials (or patients) and methods: Flow cytometric 6 in progression), 1 pt - auto þ allo-Tx. M/F 48/23; middle age (FCXM) and complement dependent cytotoxicity (CDC) cross- of pts - 53,1 y. (34-69); 15pts 60 y., 3pts 40 y. Time from matches and panel reactive antibody (PRA) analyse were diagnosis to the T# – mediana 6,5 m (2,5-95,9). Mediana follow performed pretransplant HSCT pediatric recipients. up - 26,7 m. (3,1-126,2). Early mortality (before reconstitution Results: Crossmatch and PRA tests were performed 52 hematopoiesis) – 0, 100-days mortality – 2 (2,8%). Mediana OS patients who receiving stem cells from unrelated (10/10 and PFS after Tx - 45,2 m. and 19,6 m. respectively. Mediana OS matches, n ¼ 6; 9/10 matches, n ¼ 13) and related (10/10 before 2010 - 32 m. (n ¼ 34), after 2010 – didn’t achieve matches, n ¼ 23, haploidentic n ¼ 10) donors. All of the (n ¼ 37). Since 2010 26 pts received posttransplant main- CDC crossmatch tests were negative. In 15% (8/52) of tanence therapy with bortezomib (btz) 1,3 mg/m2 s.c. one patients with T FCXM and in 36% (19/52) B FCXM positivity beweekly.

S597 Results: Since Nov. 2011 monitoring MRD were performed 26 AB103 pts. MRD detected by multiparametric flow cytometry (MFC). Preemptive low-dose interleukin-2 therapy in patients Bone marrow aspirate researches after induction therapy showing elevated minimal residual disease after (4-6 VDD or VCD); before Tx; at 3, 6, 9, 12 m after Tx in 1-st year; allogeneic stem cell transplantation: a single center study and every 6 m in the sequel. 10 pts had immunopheno- Y. Luo1,*,Y.Wu2, Y. Tan2, H. Huang2 typing remission (IFR) including 8 pts with immunochemical 1Bone Marrow Transplantation Center, the First Affiliated remission (ICR), i.e. sCR, before Tx. This pts received Hospital, Zhejiang University School of Medicine, 2Bone Marrow posttransplant therapy btz one beweekly and saved response Transplantation Center, the First Affiliated Hospital, Zhejiang (2pts achieved ICR) during follow up (4-28 m after Tx). 16 pts University School of Medicine, Hangzhou, China had VGPR and PR without IFR before Tx. This pts received btz weekly after Tx. If the paraprotein or MRD increase, the Introduction: Leukemia relapse is one of the leading cause of dexamethasone 40 mg po was added. In this group 2 pts had death after allo-HSCT. To determine the efficacy and safety of clinical relapse of the MM in 9 and 12 m., 8 pts achieved sCR low-dose Interleukin-2(IL-2) administered to patients with (IFR þ ICR) after Tx and keep response at median 18 m., 6 pts elevated minimal residual disease (MRD) after allogenetic save PR. Thereby, MRD detecting permit to define our hematopoietic stem cell transplantation(allo-HSCT) and to treatment in future. evaluate the survival benefit of this therapy. Conclusion: On our data, MRD monitoring is useful tool, which Materials (or patients) and methods: We studied 37 patients helps to determine prognosis and to choose of the who underwent low-dose IL-2 treatment between August 1, posttransplant strategy in pts with MM. However, longer 2010 to January 31,2014 at the first affiliated hospital of period of follow up and more pts in further trials are needes to Zhejiang University. Follow up to March 31,2014,we compared choose the optimal variants and duration of posttransplant the differences of outcome between groups classified by treatment. genius morbid and MRD levels. Also, we explored the safety of Disclosure of Interest: None declared. IL-2 administration. Results: The 3-year overall survival(OS), relapse-free survi- AB102 val(RFS) and cumulative occurrence rates were 83.4% (95%CI Is there an optimum CD3 þ T cell dose for donor 76.5-90.3%), 78.5%(95%CI, 71.2%485.8%)and 26.5% (95%CI, lymphocyte infusion? 18.8%434.2%). Patients stratified in the high-risk group had a S. Civriz Bozdag1,*, E. Kircali2, P. Ataca1, E. Atilla1, M. Bay1,S.K. lower OS (RR ¼ 0.122, P ¼ 0.048) and higher cumulative Toprak1, M. Kurt Yuksel1, P. Topcuoglu1, O. Arslan1, M. Ozcan1,T. occurrence rates (RR ¼ 0.036, P ¼ 0.006). Patients with higher Demirer1, O. Ilhan1, H. Akan1, M. Beksac1, N. Konuk1, G. Gurman1 MRD before the treatment of IL-2 were easier to relapse 1ANKARA UNIVERSITY DEPARTMENT OF HEMATOLOGY, 2ANKARA (RR ¼ 3.926, P ¼ 0.011.Low-dose IL-2 administration was safe. UNIVERSITY DEPARTMENT OF INTERNAL MEDICINE, ANKARA, No serious treatment- related adverse events were observed. Turkey Conclusion: Preemptive low-dose interleukin-2 therapy could be an effective and safe strategy to eliminate minimal disease Introduction: Donor lymphocyte infusion (DLI), is a salvage and control its recurrence for the patients showing elevated treatment option for patients with relapsed hematological MRD after allo-HSCT. Large clinial trials are needed in the future. malignancies after allogeneic hematopoietic stem cell trans- Disclosure of Interest: None declared. plantation (AHSCT). The graft versus tumor effect can be overwhelmed by morbidity and mortality of graft versus host disease. It has been known that diagnosis has an impact on GVHD outcome after DLI. In this study, we aimed to determine Paediatric issues the effect of the infused CD3 þ T cell dose on response, GVHD and overall survival (OS) of hematological malignancies after AHSCT. Materials (or patients) and methods: We retrospectively AB104 evaluated 56 patients with different hematological malignancy Outcome of pediatric hematopoietic stem cell diagnosis, treated with 77 DLI procedures for relapse after transplantation for inborn errors of metabolism: Single allogeneic HCT from June 2000 through January 2014. Initial center experience DLI CD3 þ cell dose/kg recipient body weight wasr1x10e7 M. Behfar1, A. A. Hamidieh1,*, S. Basirpanah1, A.-S. Hosseini1, (n ¼ 20; Group 1), 41.0 tor5 x10e7 (n ¼ 28; Group 2), 5-10 A. Ghavamzadeh1 x10e7 (n ¼ 29; Group 3). Chi-square test was used in 1Tehran University of Medical Sciences, Hematology, Oncology comparison between groups. Po.05 was considered statisti- and Stem Cell Transplantation Research center, tehran, Iran, cally significant. Islamic Republic Of Results: In 56 patients (37M/19 F), the median age during DLI was 34 (range 16-67 years). The median time interval from HCT Introduction: Inborn errors of metabolism (IEMs) are a rare to DLI was 8.06 (2-69) months. All of the patients received group of hereditary disorders that are progressive and transplants from HLA-matched related donors. Ten patients multisystem, usually is associated with early childhood death. (18%) had bone marrow while 46 patients (82%) had Hematopoietic stem cell transplantation (HSCT) has been peripheral blood as stem cell source. There was no difference established as an effective therapy for variety of IEMs. between the three groups according to age, diagnosis, stem Materials (or patients) and methods: This is a retrospective cell source and conditioning regimen.Thirty one (55%) patients study of 33 pediatric patients affected by IEMs, who had achieved complete remission after DLI. Disease status after DLI, undergone HSCT in our s- 9yecenter between 2007 and 2013. acute and chronic GVHD rates were not statistically different The median age at transplantation was 27months (range: between the groups. Overall survival according to initial DLI 5monthars). Twenty one patients were male. Patients under- cell dose at 2 years were 25%, 40% and 24% respectively, went transplantation from HLA-identical sibling donors (n ¼ 12), which was also not statistically significant (P ¼ .610). full matched other related donors (n ¼ 11), mismatched Conclusion: In previous reports,an initial DLI CD3 þ cell dose unrelated donor (n ¼ 4), matched unrelated donor (n ¼ 3) and of 10x10e7 or higher has been shown not to decrease the risk HLA-haploidentical related donor (n ¼ 3). Sources of graft were of relapse, improve overall survival but increase GVHD rates. from peripheral blood (n ¼ 14) or bone marrow (n ¼ 14) and We have compared three different DLI dose groups in our cord blood (n ¼ 5) of a healthy donor. All patients received study and found similar response, GVHD and survival myeloablative conditioning regimen with Busulfan and Cyclo- outcomes. phosphamide with or without Antithymocyte globulin. All Disclosure of Interest: None declared. patients were protected against graft versus host disease

S598 (GvHD) by infusion of cyclosporine, plus short course of count420 109 was 14 days (56 pt’s). Acute graft-versus- methotrexate except for the patients with cord blood donors. host disease of grade II to IV was observed in 69% of patients Results: Engraftment occurred in 29 0ut of 31 patient who and chronic graft-versus-host disease in 53% of patients. At were alive until 15 days after HSCT, the day of first assessment present 80 pt’s are alive and 9 pt’s died due to VOD, of chimerism. Two patients experienced secondary graft failure hemorrhagic stroke and relapse. With a median follow-up of during follow up. At the present time, 22 patients with the 18.5 months (2-30 months) after transplant the two years median follow-up of 30 months (range: 12-63 months) are still overall survival were 86.14% and event-free survival was alive and 20 of them (17 full chimerism and 3 mixed 84.1%.In Hodgkin’s disease patient’s overall survival and event chimerism) have no evidence of disease progression. Eight free survival after autologous SCT better than neuroblastoma. patients had grade III-IV acute GvHD that all of them had good Conclusion: Autologous SCT can lead to durable remissions in response to therapy. Limited chronic GvHD was developed in children and adolescents with Hodgkin’s disease & solid only one patient. The most common causes of death were tumor. These results indicate that despite our ward is new infection and disease progression. status both allogeneic and autologous HSCT are feasible with Conclusion: These results suggest that HSCT using myeloa- outcomes similar to developed countries. These preliminary blative conditioning regimen and HLA-matched related donors data suggest that HSCTs have been used as one of the can be a good combination for treatment of IEMs especially if standard treatments for hematological diseases and malig- performed early in the course of disease. Also early diagnosis nancies in Iran. and HSCT is important for a better outcome in these patients. Disclosure of Interest: None declared. Disclosure of Interest: None declared. AB106 AB105 Bone Marrow Transplantation in a child with Hematopoietic stem cell transplantation in Mahak (NGO) mitochondrial neurogastrointestinal encephalomyopathy A. Hedayati Asl1,* on behalf of Mehrvar A, Tashvigi M, Fallah V, syndrome Zangooei R, Olad E, Faranoush M, Naderi A, Dinarooni P, Nejad B. Kuskonmaz1,*, S. Aytac¸ Elmaz2, B. Konuskan3, A. Yuce4, bakhsh M, Najafi E, Ranjbar H H. Topalog˘lu3,D.Uc¸kan Çetinkaya1 1Pediatric Stem cell transplantation, MAHAK, Tehran, Iran, Islamic 1Department of Pediatrics, Divison of Pediatric Bone Marrow Republic Of Transplantation Unit, 2Department of Pediatrics, Divison of Pediatric Hematology, 3Department of Pediatrics, Divison of Introduction: The Society to Support Children Suffering from Pediatric Neurology, 4Department of Pediatrics, Divison of Cancer, also known as MAHAK, was set up in 1991 as a non- Pediatric Gastroenterology-Hepatology and Nutrition, Hacettepe governmental and non-profit organization. In the past two University, Faculty of Medicine, Ankara, Turkey decades, the organization has attracted a vast public support and fulfilled a great part of its mission which is to support Introduction: Mitochondrial neurogastrointestinal encephalo- children with cancer, reduce the child mortality rate and create myopathy (MNGIE) is an autosomal recessive disorder of an appropriate environment that empowers families who have nucleotide metabolism due to TYMP gene mutations that children with cancer. Pediatric Stem Cell Transplant also is cause loss of activity of thymidine phosphorylase (TP). MNGIE used to treat many types of conditions affecting children and manifests clinically as a multisystemic disease and character- adolescent, including cancer and certain hematologic, immu- ized by severe gastrointestinal dysmotility, cachexia, ptosis, nologic and genetic disorders. The pediatric stem cell ophthalmoparesis or both, peripheral neuropathy and leuken- transplantation ward was inaugurated in Mahak hospital cephalopathy. Hematopoietic stem cell transplantation (HSCT) (Iran-Tehran) on April, 2012. is the only potentially curative treatment that can achieve a Pediatric stem cell transplant ward practice that performs sustained biochemical correction of the metabolic imbalances. aboutn30 transplants per year. All patients are kept in high- Until now only few patients who underwent HSCT have been efficiency particulate air (HEPA) – filtered, positive-air-pressure reported. – sealed rooms, and strict hand hygiene is practiced. A 14-bed Materials (or patients) and methods: A 13.5 year old male, transplant unit specially designed for the needs of patients with consanguineous parents, referred to our hospital with the undergoing a stem cell transplant. The first case was of a complaint of abdominal pain, nausea, vomiting, diarrhea and young girl suffering from ALL and transplanted from HLA- constipation episodes, weakness especially in his lower identical sibling. extremities and weight loss. In his family history, one brother Materials (or patients) and methods: We analyzed the had died at the age of 2.5 years with a diagnosis of outcome of 89 patients from a single institution who underwent mitochondrial disease. Physical examination revealed cachec- allogeneic & autologous stem cell transplantation from between tic appearance (body weight 21.5 kg; o3 p), decreased turgor 2012–2014. Eighty one of patients had peripheral blood stem and tonus, edema in the lower extremities, bilateral ptosis, cell as the stem cell source, seven of patients’ bone marrow and weakness of lower extremity, muscle areflexia of the lower in 1 patient cord blood used. The majority of patients are: extremities, hypoesthesia in stockings distribution, wide based ALL ¼ 21, Neuroblastoma ¼ 17, AML ¼ 9, Hodgkin’s dis ¼ 23 gait. Cerebral MRI revealed disseminated lesions of white Retinoblastoma ¼ 4, Ewing’s sarcoma ¼ 2, Rhabdomyosarcom matter. MNGIE syndrome was considered and the diagnosis ¼ 2, Wilm’s tumor ¼ 2, Hepatoblastoma ¼ 1, Aplastic Anemia was confirmed by the mutational analysis of the thymidine ¼ 2, Hemoglobinopathy ¼ 1, Germ cell tumor ¼ 3, phosphorylase gene (c1112T4C). Epedymoma ¼ 1, Osteopetrosis ¼ 1.The conditioning regimens Results: The patient underwent HSCT from his HLA matched used were mainly myeloablative in allogeneic transplantation. sibling. Before the initiation of conditioning regimen the Age of patients 7 month to 26 years with median age 10 years, patient was non ambulatory, had the other neurological M/F ¼ 56/33. Thirty-three patients transplanted Allo HSCT and findings mentioned above, could not feed orally due to 56 patients transplanted Auto HSCT. GVHD prophylaxis regimen gastrointestinal dysmotility and was dependent on total was cyclosporine þ Mtx in Allo HSCT. All patients engrafted. The parenteral nutrition. The characteristics and result of HSCT type of donor in allogeneic SCT includes 28 related sibling and 5 were shown on Table 1. 3 months after HSCT the patient unrelated allogeneic. experienced mild reduction of gastrointestinal complaints. Results: In allogeneic PBSCT patients’ median time to reach Significant improvement was observed in gastrointestinal absolute neutrophil count (ANC)40.5 109/L was 11 days, findings and the patient gained 1.8 kg, and became free of and the median time to platelet count420 109 /L was 13 nasogastric tube 6 months after HSCT. He was able to walk days vs 17and 21 days in Allo BM patients. In autologous and also climb up the stairs with help 9 months after HSCT. PBSCT median time to reach absolute neutrophil count40.5 Clinical assessment 1 year after HSCT revealed that, the patient 109/L was 12 days, and the median time to platelet had very rare episodes of gastrointestinal discomfort, gained

S599 2.8 kg and his motor function was improved further and photoapheresis and steroids to control a GVHD relapse. chimerism analysis show that 98% donor profile. Currently, he is alive and event free 9 months after allo TPH. Charactersitics and results transplantation Results: Results are in conclusions chapters

2 Conclusion: Severe abdominal pain associated with IADHS Conditioning regimen Busulfan (12.8 mg/kg) þ Fludarabine (150 mg/m ) and vomits is a clinical triad suggestive of VZV atypical GVHD prophylaxis Cyclosporine A þ methotrexate reactivation in immunocompromised adults preceding Source of stem cell Bone marrow exanthema. The same triad can be identified in paediatric 8 The number of nucleated cells 5.6x10 /kg patients. The IADHS etiology is unclear. CSF is usually acellular. Engraftment ( þ ) Neutrophil engraftment day Day þ 15 Severe disseminated VZV reactivation in SCT patient is a late Thrombocyte engraftment day Day þ 26 complication with dismal prognosis if treatment is not early Acute GVHD (-) Chronic GVHD (-) started. Atypical presentation requires a high degree of clinical VOD ( þ ) (mild) suspicion to establish both diagnosis and treatment with precocity. Conclusion: HSCT is the most suitable option of treatment for References: Severe abdominal pain and inappropiate anti- MNGIE which is an otherwise intractable condition, although diuretic hormone secretion preceding vericella-zoster virus clinical evidence is still very limited. The success of HSCT may reactivation 10 months after autologous stem cell transplanta- not be expected to be high due to various pre-transplant tion for acute myeloid laukaemia, Bone Marrow Transplantation morbidities in these patients. The choice of a conditioning 2005: 35: 525-27 regimen with reduced mitochondrial toxicity is important for Disclosure of Interest: None declared. safety in MNGIE. In this case, significant clinical improvement was achieved after HSCT, without severe toxicity and AB108 transplant related events. TCR alpha/beta depletion and treosulfan/melphalan Disclosure of Interest: None declared. conditioning in juvenile myelomonocytic leukemia: single center experience AB107 D. Shasheleva1,*, M. Maschan1, L. Shelikhova1, M. Maschan1, Atypical presentation of varicella-zoster reactivation in D. Balashov 1, J. Skvortsova1, E. Kurnikova1, A. Livshits1, children after stem cell transplantation G. Novichkova1, A. Maschan1 C. Fuentes Socorro1,*, B. Torres Guerola1, V. Mateu Beitia1, 1Hematopoietic stem cell transplantation, DMITRIY ROGACHEV M. D. M. Andre´s Moreno1, C. Montoya Tamayo1, CENTER FOR PEDIATRIC HEMATOLOGY, ONCOLOGY AND J. M. Ferna´ndez Navarro1 IMMUNOLOGY, Moscow, Russian Federation 1Hospital, Hospital Universitario Infantil La Fe, Valencia, Spain, Valencia, Spain Introduction: Allogeneic hematopoietic stem cell transplanta- tion is the only curative therapy for patients with juvenile Introduction: Varicella zoster virus (VZV) reactivation is a myelomonocytic leukemia (JMML). Relapse of malignant common late complication after SCT. Atypical clinical pictures disease and transplant-related mortality (TRM) account, of VZV reactivation are uncommon but well known in adults respectively, for 30-40% and 10-20% of treatment failures. but so far to our knowledge, reports in children are very We report herein on the results of HSCT in high-risk JMML scanty. We present here two pediatric cases with severe performed with Treosulfan/Melphalan-based conditioning and abdominal pain, hyponatremia and encephalitis preceding the TCRalpha/beta and CD19 depletion. vesicular exanthema appearance. Materials (or patients) and methods: Nine patients, Materials (or patients) and methods: Patient 1: 15 years old 6 male/3 female, with JMML recieved transplantation from boy suffering ALL proB in his second remission after late matched unrelated (n - 7) or haploidentical (n - 2) donors hematological relapse. He underwent a haploidentical SCT from between May 2012 and May 2014. Median age at transplanta- his father. Both, donor and recipient were IgG positive for VZV tion was 3,6(0,8-6,5) years. Six patients had mutated PTPN11 (2 pre transplant. He presented CMV reactivation and acute GVHD of them with monosomy 7), 2 pts - c-CBL, 1 pt - NRAS, 1pt - (skin and gut), which eventually evolved to overlap skin and liver monosomy 7. Conditioning regimen included treosulfan 14 g/ chronic GVHD. Eight months after transplant being on sirolimus m2/day, days -5, -4, -3, melphalan 140 mg/m2 day -2, fludarabin and imatinib and low doses of prednisone, he complained of 30 mg/m2/ day, days -6, -5, - 4, -3, -2, ATG, horse 25 mg/kg, days severe abdominal pain. Twelve hours later, he presented a -9,-10. Peripheral blood progenitor cells were used in 8 pts, comitial crisis and went transferred to his local hospital. He was bone marrow in 1 pt. Depletion of TCRalpha/beta T-cells and CD diagnosed of IADHS (NA 128 mEq/L) and was afterwards 19 cells was performed with the CliniMACS Plus device admitted in our center ICU due to acute hypoxemic respiratory according to the recommendations of the manufacturer. Eight failure. He required mechanical ventilation. His chest X-ray was patients recieved additional post-transplant immune suppres- compatible with viral pneumonitis; a MRI of his brain did not sion with tacrolimus till day þ 30 (n - 3), tacro þ short Mtx (n - 3) show any abnormal images. Seventy-two hours later a typical or short Mtx (n - 2). Six patients recieved prophylactic post- disseminated VZV exanthema appeared. PCR was positive for transplant chemotherapy with decitabine (5) or azacytidine (1). VZV DNA in a vesicular sample. He was put on IV acyclovir and Results: In eight (88%) patients primary neutrophil and unspecific immunoglobulin. Outcome was favorable. Currently platelet engraftment was achieved at a median of 20 days. he is alive, event free, 17 months after haplo TPH. One patient failed to engraft and coud not be salvaged with Patient 2: 10 years old boy suffering AML on his second second transplant. Acute graft-versus-host disease (GVHD) complete remission after a late combined relapse. He underwent grade Y2 developed in 2 (25%) pts, chronic GVHD (de novo) an allogeneic SCT from an identical MUD. He required steroid in 1 (12,5%) pt. Relapse of JMML was registered in 6 (75%) treatment to control aGVHD (skin and gut), d þ 40. On d þ 168, patients at a median of 4,3 (1,5 -13) months. All six relapsed he complained about severe abdominal pain with vomits and a patients recieved second transplant (1 - same donor, light fever. Imaging studies were negative and no sings of 5 -alternative). At last follow-up 4 patients remain in infection were identified. Severe hyponatremia with IADHS remission (2 - after original, 2 - after second transplantation). criteria was the only analytical finding. Three days later, once his Two patients died of disease progression. Cumulative inci- natremia was back to normal, he presented a comitial crisis. MRI dence of relapse and TRM are 78% (95%CI:45-100) and 0%. was normal and CSF was acellular with no biochemical Event free survival is 22% (95%CI:0-49) and overall survival abnormalities. On the forth day a typical vesicular exanthema 67% (95%CI: 36-97) at 2 years. appeared. PCR for VZV DNA was positive both in CSF and skin Conclusion: Although limited to few observations and biased lessions. The outcome was favorable on IV acyclovir and by overrepresented unfavorable genotypes (PTPN11 and/or unspecific immunoglobulin. He required extracorporeal monosomy 7), this analysis suggests that TCRalpha/bate

S600 depleted transplantation in combination with treosulfan/ was bone marrow in 192, mobilised peripheral blood stem cells melphalan/fludarabine conditioning do not provide optimal in 123, cord blood in 41 and combined bone marrow and anti-leukemic activity in JMML. peripheral stem cell in 7 patient. Transplant characteristics are Disclosure of Interest: None declared. shown in table 1. The Kaplan-Meier estimate of OS and EFS at one year were 74%±3, 67%±3, respectively with a median one year follow up. One year OS was significantly inferior for malign AB109 diseases compared to non-malign ones ( 60%±4vs83%±3, Busulfan pharmacokinetics in Turkish children who Po0.001). A graft from MSD was associated with a best OS, underwent hematopoetic stem cell transplantation although the difference with matched related donor did not E. O¨ zyu¨rek1,2,*, B. Karagun3, B. Antmen3, I. Sasmaz3,4, M. Serbest3, reach to statistical significance ( 87%±3 for MSD, 77%±6for G. Ucar3, Z. Guney2, E. Sahin2, M. Oktem5, T. Fisgin1,2 MRD, 61%±5 for MUD). One year OS was superior in patients 1Pediatrics, Bahcesehir University, School of Medicine, Istanbul, receiving bone marrow as a stem cell source compared to 2 Pediatric Bone Marrow Transplantation Unit, Samsun Medical- peripheral stem cell (80%±3vs63%±5, P ¼ 0.005) In a park Hospital, Samsun, 3Pediatric Bone Marrow Transplantation multivariate cox model, nonmalign disease and also graft from Unit, Acibadem Adana Hospital, 4Pediatric Hematology, Cukur- sibling donor was associated with better OS. ova University, School of Medicine, Adana, 5Duzen Laboratuvar Grubu, Ankara, Turkey Table 1 Characteristics of tranplantations

Introduction: Busulfan is a cytotoxic drug usually used as a n (%) part of conditioning regimen in hematopoetic stem cell transplantation. Individualised dosing of busulfan by measure- Number of total transplants 362 Allogeneic 344 ment of its pharmacokinetics, and adjusting its dose, reduced Autologous 18 its major side effect, namely venoocclusive disease. No of patients 347 Materials (or patients) and methods: We retrospectively Indications Thalassemia 79 ( 22.8) evaluated pharmacokinetics of intravenous busulfan in 79 Turkish Acute lymphoblastic leukemia 75 (21.6) children with a median age of 7 years (2-20 years) who under- Acute myeloid leukemia 37 (10.7) went hematopoetic stem cell transplantation for various hemato- Primary immune deficiency 47 (13.5) Bone marrow failure 39 (11.2) logical diseases. Intravenous busulfan was administered every Inborn error of metabolism 25 (7.2) 6 hours as a part of a conditioning regimen with pharmacokine- Lymphoma 19 (5.5) HLH 11 (3.2) tically adjusted dose to target an area under the concentration- MDS 9 (2.6) versus-time curve (AUC) range (900-1350 mMol*min). Solid tumor 6 (1.7) Results: Among them, 50 was transplanted for thalassemia, 16 Donor type MSD 160 for acute lymphoblastic leukemia, 8 for acute myeloid leukemia, MRD 49 and 5 for various metabolic and hematological disorders. The MUD 118 MMRD 17 median AUC of the initial dose was 912 mMol*min (342-1320 Stem cell source mMol*min). 38 patients achieved the target range AUC, 29 BM 192 patients required dose escalation, whereas 12 needed dose PBSC 123 CB 34 reduction. The mean initial busulfan dose was 0,9 mg/kg/dose CB þ BM 7 (0,7-1,2 mg/kg/dose), and the mean busulfan dose after BM þ PBSC 6 adjusment was 1 mg/kg/dose (0,8-2,4 mg/kg/dose). Conclusion: In conlusion, the results of busulfan pharmaco- HLH; hemophagocytic lymphohistiocytosis, MDS; myelodys- kinetics of Turkish children who underwent hematopoetic plastic syndrome,MSD;matched sibling donor, MRD; matched stem cell transplantation for various hematological diseases related donor, MUD; matched unrelated donor, MMRD; are comparable to previously performed studies. mismatched related donor, BM; bone marrow, PBSC; periph- Disclosure of Interest: None declared. eral blood stem cell, CB; cord blood. Conclusion: These data reflect the present status of HSCT in AB110 Turkey. Due to high incidence of consanguinity between A retrospective review of the outcome after hematopoetic parents, the number of patients who underwent transplanta- stem cell transplantation in children tion with the indication of inherited diseases were high. With incremental experience, the patient group has diversified G. Karasu1,*, S. Caki Kilic1, O. Ozboru1, F. Pekun1, A. Yesilipek1 1 towards more advanced patient status and to patients without Pediatric Stem Cell Transplantation Unit, Bahcesehir University a family donor and the outcomes are comparable to that Faculty of Medicine Goztepe Medicalpark Hospital, Istanbul, reported in other studies. Turkey Disclosure of Interest: None declared. Introduction: The aim of this study was to document hematopoietic stem cell transplantation activity and examine AB111 outcome in a newly established pediatric stem cell transplan- Abstract Withdrawn tation unit. Materials (or patients) and methods: The data for this analysis was taken for the timespan April 2011, the date when our transplantation unit was established, to November 2014 AB112 and retrospectively analyzed, concentrating on types of HSCT, Successful maintenance treatment with interleukin-2/ indications for transplantation, donor types, stem cell sources histamine dihydrochloride (IL-2/HDC) and after and causes of death following transplantation. Both overall haploidentical SCT for refractory relapse of AML J. Greil1,*, J. Kunz1, P. Lang2, M. Claus3, A. Kulozik1 (OS) and progression-free survivals (PFS) were estimated by 1 Kaplan-Meier analysis. Pediatric Hematology and Oncology, UNIVERSITY HOSPITAL HEIDELBERG, Heidelberg, 2Pediatric Hematology and Oncology, Results: In total, 347 patients received 362 transplantation 3 procedures in our center, 18 of which was autologous (auto) University Hospital Tuebingen, Tuebingen, Working Sciences, transplant.The most common indication for HSCT was hemo- Leibnitz-Institute, Dortmund, Germany globinopathy followed by leukemia. The most frequent stem cell donor was an identical sibling (n ¼ 160), followed by an Introduction: Allogeneic hematopoietic SCT is highly effective unrelated donor (n ¼ 118), a matched relative (n ¼ 49) and for cure of chemotherapy sensitive AML relapse. But cure rates finally a mismatched family donor (n ¼ 17). The stem cell source for chemotherapy refractory AML relapse are remaining low.

S601 Materials (or patients) and methods: A 15-months-old girl 3,29-25,21); 6,52 106/kg CD3 þ (0,031-19,06); 593,67 103/ was diagnosed with first extramedullary relapse of AML. The kg CD20 þ (0-1913,08); and 16.646,82 103/kg NK cells patient’s initial acute myeloid leukemia had shown M4 (6.462,53-24.281,43). Graft versus host disease (GVHd) prophy- morphology and a complex MLL-rearrangement. Relapse laxis included mycophenolate mofetil in 1 patient and treatment with liposomal daunorubicin and fludarabine was cyclosporine and metothrexate (days 1, 3 and 6) in 4. initiated, but the malignant skin lesions were refractory to Results: All patients achieved primary engraftment; however, chemotherapy. At the age of 18 months the girl was 2 patients (1 ES and 1 NBL) experienced secondary graft failure transplanted with CD3/CD19 depleted peripheral blood stem and were rescued with subsequent allogeneic stem cell cells from her haploidentical mother. Conditioning regime transplantation from a matched sibling donor (MSD). One consisted of fludarabine (40 mg/m2/d x 4), thiotepa (10 mg/kg), patient had severe (grade 4) steroid resistant acute GVHd melphalan (70 mg/m2/d x 2) and OKT3 (0.1 mg/kg/d x 28). responding to extracorporeal photopheresis. Three patients Results: The malignant skin lesions disappeared completely had disease progression and died of disease 5 to 12 months after development of GvHD I1 of the skin. On d þ 50 s.c. after haplo-SCT (23 to 35 months after primary diagnosis). The injections with IL-2/HDC were started. Within 27 months 2 patients with ES are currently alive and disease-free after 12 overall fifteen 21- day cycles with IL-2 (16,400 U/kg/d, s.c.) plus and 36 month of follow up after haplo-SCT (44 and 57 months HDC (0.01 mg/kg/d, s.c.) were applied. Only moderate side from primary diagnosis). effects, i.e. local pain and transient rash occurred. IL-2/HDC Conclusion: In our experience, T-cell depleted haplo-SCT is treatment induced an increase of NK-cells and an enhanced feasible in pediatric patients with systemic relapse of cytotoxic activity of the patient’s NK-cell against K562-cells. aggressive ST with no death directly related to the Twelve months after haploidentical SCT a second extramedul- procedure. Graft rejection occurred in 2 patients with lower lary relapse with a single skin lesion on the right thigh numbers of CD3 infused (o 0,2 106/kg); both were occurred. This skin lesion was resected with safe margins. rescued with a subsequent transplantation from a MSD, Treatment with sorafenib in a dosage of 200 mg/m2/d was and one of them is alive and disease-free 44 months started and this dosage was continued for overall 30 months. after a systemic relapse of ES. Only patients that underwent Sorafenib treatment was tolerated well. After slow tapering of haplo-SCT in CR2 are currently alive and desease free, sorafenib the patient is remaining in third continuous underscoring the role of disease status before transplantation remission for now more than 44 months. for treatment success. Conclusion: This case shows that post-transplant mainte- References: nance treatment with IL-2/HDC and sorafenib after haploi- . Leung W. Infusions of allogeneic natural killer cells as cancer dentical SCT is effective. Moreover it also indicates that therapy. Clin Cancer Res. 2014 Jul 1;20(13):3390-400. prospective studies with post-transplant maintenance treat- . Brehm C, Huenecke S, Pfirrmann V, Rossig C, Mackall CL, ment should be performed. Bollard CM, Gottschalk S, Schlegel PG, Klingebiel T, Bader P. Disclosure of Interest: None declared. Highlights of the third International Conference on Immu- notherapy in Pediatric Oncology. Pediatr Hematol Oncol. 2013 AB113 Aug;30(5):349-66 T-cell depleted haploidentical stem cell transplantation for Disclosure of Interest: None declared. systemic relapse of aggressive pediatric solid tumors J. L. Fuster-Soler1,2,*, J. Monserrat-Coll1,2, J. A. Campillo- AB114 Marquina1,2, M. Blanquer-Blanquer1,2, A. Minguela-Puras1,2, A successful Hematopoietic Stem Cell Transplantation in A. Sa´nchez-Salinas1,2, A. Fita1, J. F. Pascual-Ga´zquez1,2, Wiskott-Aldrich syndrome using a mieloablative regimen A. Galera-Min˜arro1,2, M. E. Llinares-Riestra1,2, L. Sisinni1,2,*, M. Torrent 1,2, N. Pardo1,2, E. Fernandez3, T. Toll4, M. Bermu´dez-Corte´s1,2, J. M. Moraleda1,2 O. De la Calle5, I. Badell 1,2 1Stem cell transplantation and cell therapy unit, HOSPITAL 1Pediatric BMT Unit, Hospital Santa Creu i Sant Pau, 2Pediatrics, CLI´NICO UNIVERSITARIO VIRGEN DE LA ARRIXACA, 2IMIB, Universitat Auto`noma de Barcelona, 3Farmacology, Hospital Universidad de Murcia, Murcia, Spain Santa Creu i Sant Pau, 4Pediatric Hematology/Oncology, H.Sant Juan de Deu, 5Inmunology, Hospital Santa Creu i Sant Pau, Introduction: Advanced diseases or systemic relapses of Barcelona, Spain aggressive childhood solid tumors (ST) such as Ewing sarcoma (ES) have very poor prognoses. Haploidentical hematopoietic Introduction: Hematopoietic stem cell transplantation (HSCT) stem cell transplantation (haplo-SCT) has become a feasible is the current accepted curative approach for patients with and safe procedure with potential efficacy in childhood Wiskott-Aldrich syndrome (WAS). The reduced-intensity con- malignancies. In haplo-SCT, alloreactive NK cells may induce ditioning regimens have still an increased risk of graft rejection potent antitumor effects, which can be predicted by KIR/KIR- compared to myeloablative approaches. ligand genotyping of donor/recipient. Materials (or patients) and methods: We describe a case Materials (or patients) and methods: From november 2011 report of a one year old male affected by classic WAS. The we offered haplo-SCT on a compassionate basis to 5 pediatric diagnosis was made for the presence of thrombocytopenia patients (age 6 to 15 years) with disseminated relapse of ES (2), with low mean platelet volume, immunodeficiency (B-cell and neuroblastoma (NBL, 2) and osteosarcoma (OST, 1). Three T-cell lymphopenia, very low IgM) and eczema. The molecular patients (2, ES and 1 OST) underwent haplo-SCT in second study demonstrated a ‘‘de-novo’’ mutation. The patient was remission (CR2) after surgical removal of lung metastases and submitted to an allogeneic HSCT from an unrelated donor with second line chemotherapy; the 2 remaining patients pro- HLA identity. A reduced intensity conditioning regimen based ceeded to the haplo-SCT with progressive systemic disease on fludarabine, melphalane and alemtuzumab was used. The (bone and/or marrow infiltration). Conditioning regimen GVHD prophylaxis was based on cyclosporine and mofetil consisted of busulphan (Bu), fludarabine (Flu), metilpredniso- mycophenolate. After 2 years, because of the loss of the lone (MP) and thiotepa (TT) in 4 patients and radioactive meta- engraftment, he received a second bone marrow transplanta- iodo-benzil-guanidinine followed by Flu, TT, melphalan and tion from an other HLA- total matched unrelated donor. A anti-thymocyte globuline in 1. In all except 1 patient, donor myeloablative conditioning regimen based on busulfan selection was based upon KIR and KIR-ligand genotyping of (adjusted dose), cyclophosphamide and rabbit ATG was used. both receptor and donor and on donor KIR expression by flow; Cyclosporine was given for the GVHD prophylaxis. donor selected was the mother in 3 patients and the father y 2. Results: Total chimerism was achieved after the first Ex vivo manipulation consisted of CD3/CD19 depletion in 2 transplantation and the laboratory analysis confirmed pro- patients, and TCRab/CD19 depletion in 3. The grafts contained gressive normalization of platelets level. After 1 year the a mean of 11,24 106/kg of recipient weight CD34 þ (range: chimerism was progressively lost with final rejection

S602 of graft in myeloid lineage. At the same time platelets started AB116 decreasing achieving pre-trasplant levels (o20,000/mL). The Effect of Levetiracetam on Electroencephalogram After the second transplant full and stable engraftment Abnormalities That is Induced by Busulfan in Children was achieved (in myeloid and lymphoid compartments) T. Cokyaman1, M. Elli2,*, O. F. Aydin1, C. Albayrak3, D. Albayrak3, and has been mantaining during the 24 months after T. Fisgin4, E. Ozyurek2,5 transplantation with normal and stable platelet level. We 1Department of Pediatrics, Divsion of Pediatric Neurology, observed complete immunological recovery at 2 years after Ondokuz Mayis University Faculty of Medicine, 2Department of transplant. Pediatrics, Division of Pediatric Hematology, Pediatric BMT/HCST Conclusion: The type of the conditioning regimen is an Unit, Samsun Medical Park Hospital, 3Department of Pediatrics, important issue in the care of a patient with Wiskott-Aldrich Division of Pediatric Hematology, Ondokuz Mayis University syndrome who will be submitted to HSCT. In our Faculty of Medicine, Samsun, 4Department of Pediatrics, Division report total engraftment was achieved after both trans- of Pediatric Hematology, Bahcesehir University, School of plantations nevertheless only with a myeloablative preparative Medicine, 5Department of Pediatrics, Division of Pediatric regimen stable engraftment was maintained showing a Hematology, Bahcesehir University School of Medicine, Istanbul, normalization of platelet level and complete immunological Turkey recovery. Disclosure of Interest: None declared. Introduction: Busulfan (BU) is a neurotoxic chemotherapeutic agent, commonly used in anticancer treatment and bone AB115 marrow/hematopoietic stem cell transplantation (BMT/HSCT) Transfusion of peripheral blood lymphocytes from conditioning regimen. This study was designed to evaluate the HLA-identical donors to establish a functional T-cell recent electroencephalogram (EEG) abnormalities after BU system: experience in 3 pediatric patients with primary treatment and effectiveness of levetiracetam (LEV) given T-cell deficiencies prophylactically. Materials (or patients) and methods: In this study, the M. Hoenig1,*, I. Furlan1, C. Schuetz1, W. Friedrich1, A. Schulz1 1 records of patients who received BU in their conditioning Pediatrics, University Medical Center, Ulm, Germany regimen at the BMT/HSCT unit between February 2009 and January 2014 were assessed retrospectively. The diagnosis, Introduction: We present 3 patients with primary T-cell demographic characteristics, clinical seizure frequency of deficiencies, who for different reasons received peripheral patients after BU treatment, recent EEG abnormalities and blood transfusions which led to effective and stable T-cell antiepileptic prophylaxis were evaluated. reconstitution. Results: Forty-nine patients were given BU in conditioning Materials (or patients) and methods: Data on three regimen. LEV was given to 63% and phenytoin (PHT) to 36% of consecutive patients with T-cell deficiency treated with patients as antiepileptic prophylaxis. Clinical seizures were peripheral blood leukocytes were collected retrospectively. observed in 6% of patients after BU treatment. Frequency of Results: The first patient was identified with complete seizure episodes did not differ between LEV and PHT DiGeorge Syndrome at the age of 2.5 months and received prohylaxis groups. When we compare mean age of patients, 2 transfusions of peripheral blood leukocytes from his HLA- whose EEG’s were normal and who developed EEG abnorm- identical brother, each containing a total of 10e8/kg CD3 þ alities, there was no significant difference. There was neither cells. This led to persistent donor T-cell reconstitution (CD3 þ statistically significant association of thalassemia major (TM) cell counts 800-1600/ml) without any serious infections within with EEG abnormalities following BU therapy, nor statistically a follow up of 12 years. All other cell lines remained of significant difference between the effects of LEV and PHT on autologous origin. these abnormalities. The second patient was diagnosed with ZAP-70 deficiency at Conclusion: EEG abnormalities emerging after BU and seizure birth. His HLA-identical brother with the same disorder had frequency were not correlated with patient’s age and disease previously been transplanted successfully from a MUD after specifically TM. LEV and PHT had comparable effects in conditioning. The patient received 2 transfusions of peripheral preventing seizures and EEG abnormalities following BU use in blood (CD3 þ 12x10e6/kg each) from this brother. In addition BMT/HSCT conditioning regimens. a CD34 þ selected, kryopreserved stem cell graft from the Disclosure of Interest: None declared. previous donor of his brother was transfused. No conditioning was given. The patient developed effective T-cell functions (CD3 þ cell counts 1000/ml). Naive donor T cells were never AB117 detectable and all non-T-cell populations remained of auto- A 10-year retrospective study of Hematopoietic Stem Cell logous origin. The patient has not experienced any serious Transplantation for cancer and benign hematologic infections within a follow up of 6 years. diseases: A single report from a Mexican Pediatric The third patient was diagnosed with JAK-3 deficiency at the Hematology and Oncology Center age of 5 months presenting with pneumonia, encephalitis O. Gonzalez-Ramella1,2,*, J. L. Vazquez-Urdiano3, M. M. Ortiz- and myocarditis caused by CMV. In spite of i.v. ganciclovir Sandoval2,4, A. L. Orozco-Alvarado 2,4, R. M. Navarro-Martin Del her viral load was not reduced. She received 1,6 ml of Campo2,4, G. Escobedo-Melendez3,4, F. Sanchez-Zubieta3,4, peripheral blood from her HLA-identical mother, who was R. Balderrama-Magallanes5 CMV IgG positive, with a T-cell content of 0.5x10e6/kg 1Instituto de Investigacion del Cancer infantil y de la CD3 þ . This led to rapid appearance of circulating donor T Adolescencia, Universidad de Guadalajara, 2Bone Marrow cells and subsequent clearance of the viral infection. In Transplantation Unit, 3Servicio de Hematologia y Oncologia association with a mild cutaneous GvHD, the patient devel- Pediatrica, Hospital Civil de Guadalajara "Juan I Menchaca", oped aplastic anemia 4 weeks after the transfusion. A T-cell 4Instituto de Investigacion del Cancer infantil y de la Adolescen- depleted maternal stem cell graft led to complete donor cia, 5Medicine student, Universidad de Guadalajara, Guadalajara, chimerism and full immunological reconstitution. She is Mexico thriving well with lower limb motor dysfunction after CMV encephalitis and has remained free of serious infections within Introduction: In children, hematopoietic stem cell a follow up period of 3,5 years. transplantation (HSCT) plays an important role in the Conclusion: Our experience demonstrates the potential of treatment of malignant and not malignant diseases. In our infusions of mature peripheral blood T cells from HLA-identical hospital, the first HSCT was performed in 2003. Currently, the donors to rapidly establish a long lasting functional T-cell pediatric Hematopoietic Stem Cell Unit (HSCU) has been system in primary T-cell deficiencies. working for 11 continuously years attending patients from Disclosure of Interest: None declared. western and north Mexico. The aim of this study was to

S603 evaluate the characteristics and the outcome of HSCT in our because of uncooperation of patients or difficulties Center. related to the age. The characteristics of the patients Materials (or patients) and methods: Between January 2003 are described in the following table. TBI has been adminis- to December 2013, 95 pediatric patients under the age of 20, tered by a 15 MV linear accelerator in twice a day with either a malignant or non-malignant diseases were fractions of 2 Gy for a total of 12 Gy, in supine position with transplanted in our Unit. This hospital is classified as a tertiary LL technique and with a lead protection to reduce pulmonary care Center and the patients attended have a low socio- dose to a total value of 9 Gy. Lead shields were also used also economic status. Clinical records were reviewed and data were to protect other sensitive organs to compensate dose increase retrospectively analyzed. and mantain 12 Gy total dose. Dose was monitoring during Results: A total of 95 pediatrics patients were trans- treatment with in vivo diode dosimetry. The children have planted. Fifty-three (55.8%) were male children; average age been sedated according to a protocol which provided for a was 7.6 years (range 1 to 20). Twenty-two patients (23.2%) pre- treatment with Ondansentron, Ranitidine, Desametasone were diagnosed with acute lymphoblastic leukemia, 24 and midazolam (1-3 mg total dose), administration of keta- (35.3%) with acute myeloid leukemia, 31 (32.7%) with mine (1-2 mg/kg i.v.), continuous cardiorespiratory monitoring benign hematologic disorder (including immunodeficiencies), and O2 in mask. An anaesthetist-resuscitator is always present 4 (4.3%) with solid tumors, 3 (3.2%) with myelodysplastic in the room. Furthermore, in the operating room there is a syndrome, 2 (2.1%) with lymphoma, and 9 (9.5%) with camera, which allows to monitor the children during the granulocytic chronic leukemia. Fourteen patients (14.7%) session. received an autologous transplant, a full matched related Results: donor (MRD) was used in 49 (51.5%) whereas 32 (33.6%) a full or partially matched unrelated donor (MUD) was used, (4/6 to 6/6 30 cord blood units and 1 haploindentical). N. Diagnosis Age RT Position Sedation Different conditioning regimens were used including myelo- and non-myelobaltive. Total body irradiation was used 1 ALL 12 y (Down Syndrome) Supine Midazolam/Ketamine/O2 in all patient with an ALL. Average successful graft was 2 ALL 6 y Supine Midazolam/Ketamine/O2 3 ALL 4 y Supine Midazolam/Ketamine/O2 achieved in 21.6 days (range 8 to 71). Some grade of acute 4 ALL 3 y Supine Midazolam/Ketamine/O2 GVHD occurred in 33 patients (34.7%) whereas chronic GVHD was seen only in 3 (3.1%). Overall survival rate was 55% with 60 follow up time. No one of the patients has shown neither remarkable Conclusion: HSCT is feasible in low- and mid-income countries collateral effects which could be referred to the anesthesia, like ours. MUD program was initiated in 2008 in our Hospital, nor problems which have impeded the regular administration most of them using a cord blood unit. We used for all MRD of the TBI or condiotining. The median duration of radio- bone marrow as a stem cell source in contrast to other centers therapy session is 10 minutes and the children is sedated for in Mexico; that is reflected in our very low incidence of cGVHD. 15-20 minutes. We didn’t observed any problems with drugs Both neutrophil and platelet engraftment time were longer used for sedation. The dose delivered during each treatment compared to other HSCT Centers, this might be explained due was within 2% of the prescribed value. to the high number of cord blood units that were transplanted A patient is deceased because of an infection at the day þ 5 as well as the use of bone marrow instead to peripheral blood. from the TMO; in the other 3 cases, it has been registred a Relapse and infections are still the major cause of death after regular engraftment of the graft HSCT. Transplant related mortality has been decreased over Conclusion: Recently it is increasingly the use of preparatory the years, due principally to the multidisciplinary team regimens TBI-free, however the whole body irradiation expertise. We are working together with the majority of remains an important part in the conditioning of allogeneic pediatric HCSU in Mexico in order to report the real HSCT bone marrow transplants especially when executed for ALL. situation in our country. However, not all radiotherapy units are able to perform TBI- Disclosure of Interest: None declared. sessions under anesthesia. It is frequent that there are unsolvable logistical difficulties when patients are uncoopera- tive or very young children. AB118 When necessary, the total body irradiation during sedation of Feasibility and effectiveness of the whole body these young patients is a feasible, safe and effective irradiation during sedation in pediatric uncooperative procedure. patients References: S. Tropia1, G. Granata2, F. Magliolo2, B. Abbate3, V. Caputo3,G. Disclosure of Interest: None declared. Evangelista4, F. Sciume`4, F. Di Marco1, C. Mosa1, A. Trizzino1,O. Ziino1,* 1ARNAS Civico e Di Cristina Palermo Italy, Paediatric Haemathol- AB119 ogy and Oncology Department, 2ARNAS Civico e Di Cristina Results of Hematopoietic Stem Cell transplantation for Palermo Italy, Anesthesia and Rianimation Service, 3ARNAS Civico High Risk Pediatric Leukemia: The experience in a e Di Cristina Palermo Italy, Medical Physics Department, 4ARNAS Single-Centre in Mexico Civico e Di Cristina Palermo Italy, Radiotherapy Department, R. M. Navarro-Martin Del Campo1,2,*, A. L. Orozco-Alvarado1,2, Palermo, Italy M. M. Ortiz-Sandoval1,2,F.A.Sa´nchez-Zubieta2,3, S. Gallegos-Castorena3, O. Gonza´lez-Ramella1,2 Introduction: Although there are evidences of the 1Unidad de Trasplantes de Progenitores Hematopoyeticos, effectiveness of the TBI-free preparatory regimens, the total Hospital Civil de Guadalajara "Juan I Menchaca", 2Instituto de body irradiation is still widely used in the conditioning Investigacion del Cancer en la infancia y la Adolescencia, of the allogenic bone marrow transplants, especially Universidad de Guadalajara, 3Servicio de Hematologia y in case of ALL. However, this procedure can present difficulties Oncologia Pediatrica, Hospital Civil de Guadalajara "Juan I in very young or uncooperative children and it must be Menchaca", Guadalajara, Mexico executed during sedation. In the table below are reported our clinical records of the TBI procedures, performed during Introduction: Hematopoietic stem cell transplantation (HSCT) narcosis. offers an opportunity to cure children with new diagnosed or Materials (or patients) and methods: At the Pediatric relapsed high risk leukemia. Prognosis in these patients is Haematology and Oncology Department of the ARNAS usually dismal with conventional chemotherapy. The purpose Civico and Di Cristina Hospital of Palermo, in the last of this communication is to describe the results in Pediatric years we performed 4 TBI procedures during narcosis, High Risk Leukemia in our Centre.

S604 Materials (or patients) and methods: Between, March 2003 transplantation with unrelated 10/10 HLA matched peripheral to May 2014, fifty nine pediatric patients with high risk stem cell. Conditioning was non myeloablative consisting of leukemia aged between 1 to 20 years underwent a HSCT. melphalan, fludarabine and alemtuzumab or ATG in all Status disease were: ALL 1st CR:10, ALL 2nd CR: 13, ALL 3rd CR: patients. Neutrophil engraftment was on þ 34, þ 32 and 4, AML: 22, CGL non responsive to imatinib:8, MMJL:1 þ 17 days respectively. No GVHD was seen. All patients are Secondary LLA:1. Conditioning regimens were myeloablative alive and in remission after 52, 19 and 13 months after in all cases. Graft Versus Host disease (GVHD) prophylaxis transplantation. consisted in cyclosporine plus methotrexate. Results: There is a paucity of data regarding the treatment of Results: Stem cell sources included: Bone Marrow (BM) in 71%, relapsed or refractory LCH. The role of allogeneic HSCT in LCH Peripheral Stimulated Blood (PSB) in 12% and Cord blood (CB) has not been directly studied. Case reports and small case in 17%. Nine patients with AML received autologous series suggest that HSCT may be effective in preventing transplantation while the rest 50 received an allogenic HSCT. relapse among patients in complete remission at the time of Medium Infused dose of CD34 cells/Kg for Bone Marrow group HSCT. Relapses or toxicity of conditioning are important risk were 3^6, in CB group 0.21^6 and in PSB group 5^6. factors for successful transplantation. All of our patients had Engraftment was achieved in 53 patients, 6 patients had graft non myeloablative conditioning regimens. We had no toxicity failure, one of them transplanted from CB. Neutrophil of the procedure. We had no adverse event associated with engraftment was achieved in average at day þ 17 in BM the source of stem cell other than relatively delayed and PSB patients while in UCB at day þ 28. Platelet recovery engraftment with cord blood. was achieved in average at day þ 20 in BM and PSB Conclusion: Allogeneic HSCT with non myeloablative con- patients while in UCB at day þ 34. Acute GVHD was diagnosed ditioning resulted in the apparent cure of all of our patients in 45% of patients. The EICH grade was I-II in 60% of after failing chemotherapy. cases and Grade IV in 40%. Mild Chronic GVHD was diagnosed Disclosure of Interest: None declared. in 10 patients. At the mean time 61% of patients are disease free with a follow up average of 39 months. Remarkably AB121 all but one autologous transplanted AML patients are alive. Post-transplantation Immune Reconstitution Syndrome in Causes of death were Leukemic progression in 52% of cases a Patient with DOCK8 Deficiency and Molluscum followed by aGVHD in 22%, CMV infection after day 100 Contagiosum in 9%, graft failure in 9%, sepsis in 4% and Veno oclussive 1,* 2 3 4 3 disease in 4%. S. Unal , S. Ersoy Evans , I. Tezcan , M. S. Boyraz , D. Cagdas ,D. Uckan Cetinkaya1 Conclusion: We found that HSCT is a viable option for patients 1 2 with high risk pediatric leukemia, reaching 61% of cure in our Hacettepe University, Division of Pediatric Hematology, Hacet- tepe University, Department of Dermatology, 3Hacettepe Uni- series, although mean cause of death post transplantation still 4 being leukemic relapse. These results are closed to reports in versity, Division Pediatric Immunology, Hacettepe University, other Centers. In our population most cases of chronic GvHD Department of Pediatrics, Ankara, Turkey were mild to moderate, we believe that it is mainly due to use BM and CB as a source of hematopoietic progenitors. Introduction: Immune reconstitution sydrome (IRIS) has Disclosure of Interest: None declared. previously been described among patients with tuberculosis after initiation of anti-tuberculosis treatment and among patients with HIV associated HPV or molluscum contagiosum AB120 infections after initiation of highly active anti-retroviral therapy Hematopoietic Stem Cell Transplantation In Pediatric (HAART) and is characterized with the transient worsening of Langerhans Cell Histiocytosis: Experience of Ege University symptoms related to recovering immune system. Herein, we From Turkey describe a patient with DOCK8 deficiency and AML who S. Gozmen1,*, S. Aksoylar1, N. Çetingu¨l2, M. Kantar2, developed IRIS in the molluscum lesions subsequent to HSCT Z. O¨ nder Sivis¸2, S. Kansoy1 at engraftment stage. 1Pediatric BMT Unit, 2Pediatric Oncology, Ege University, Izmir, Materials (or patients) and methods: An eight-year-old boy Turkey was admitted for HSCT from MSD. The previous history revealed a diagnosis of DOCK8 deficiency with recurrent Introduction: Langerhans cell histiocytosis (LCH) is a rare infections, biopsy proven anogenital and facial molluscum histiocytic disorder. The prognosis of patients with disease contagiosum related lesions, multiple food allergies and Hyper progression after a first course of chemotherapy is very poor IgE. The diagnosis of DOCK8 deficiency was supported with and hematopoietic stem cell transplantation (HSCT) may be an molecular analyses. He was on IVIG treatment when devel- option in such cases. HSCT has been proposed for LCH in oped acute myeloid leukemia at the age of seven-years. AML- various clinical situations, including recurrent multifocal bone BFM 2004 protocol was initiated, however after a course of lesions, adult lung involvement, and refractory LCH in children. brief remission, relapse occurred and FLAG-Ida protocol was Here, we present successful transplantation of three refractory given and the patient underwent HSCT afterwards. Condition- pediatric LCH patients after HSCT. ing regimen of melfalan, fludarabine and ATG was given and Materials (or patients) and methods: Patients were 7.5 methotrexate and cyclosporine A were given for GvHD months, 8 months and 9 months old at the time of diagnosis prophylaxis. Bone marrow harvested stem cells from matched of LCH. First two patients were male and the third patient was sister donor was given after erythrocyte depletion (CD34 dose: fenale. The diagnosis of LCH was based on the Histiocyte 2.13x106/kg). Society criteria. Organs involved by LCH at diagnosis were Results: Neutrophil and thrombocyte engraftments were liver, spleen, bone marrow and skin in the first two patients. achieved by day þ 17 and 23, respectively. Patient was Third patient had skin involvement. All patients had frontline observed to have exacerbation of the molluscum lesions treatment according to LCH-IV protocole with vinblastine and including erythematous enlargement of the verrucous lesions steroid. While first two patients had poor response to frontline on day þ 12 and gradually worsened before almost totaly treatment after six weeks, third patient had good response to healed on day þ 64. During the worsening period local frontline treatment but relapsed after five months of cessation ointments with antibiotic and cleansing of the lesions were of therapy. In addition, all patients had methotrexate, applied. cyclosporine, cytarabine, cladribine and cloforabine after. The Conclusion: The immunological boost after HSCT in a patient duration of LCH prior to HSCT was 13 months, 22 months and with DOCK8 deficiency and AML caused inflammatory reaction 18 months respectively. First two patients had trans- to the previously existing molluscum contagiosum virus, platation with unrelated cord blood (6/6 and 5/6 HLA matched preceeding to the healing of the lesions.The phenomenon of with 6,5x107/kg and 3,8x107/kg). Third patient had parodoxical IRIS has especially been reported among patients

S605 with HIV infection who were initiated HAART and developed Experimental stem cell transplantation transient worsening of either HPV or molluscum contagiosum lesions. Upto our knowledge, this is the first report of IRIS developed subsequent to HSCT in a patient with severe cellular immunity defects and molluscum contagiosum lesions, AB123 who developed IRIS. The physicians should be aware of the Allogeneic bone marrow and mesenchymal stem cells transient status and actually a healing process of this entity in therapy for dystrophic epidermolysis bullosa M. KARAKUKCU1, E. YILMAZ1, S. KURTOGLU2, M. A. OZDEMIR1, order to avoid unnecessary interventions which may increase 1 1,* the toxicities. T. PATIROGLU , E. UNAL 1Department of Pediatrics, Division of Pediatric Hematology References: Link-Gelles R, Moultrie H, Sawry S, Murdoch D, 2 Van Rie A. Tuberculosis Immune Reconstitution Inflammatory Oncology, Department of Pediatrics, Division of Neonatology, Syndrome in Children Initiating Antiretroviral Therapy for HIV Erciyes University, Faculty of Medicine, Kayseri, Turkey Infection: A Systematic Literature Review. Pediatr Infect Dis J. 2014;33:499-503. Introduction: Epidermolysis bullosa (EB) represents a group of Disclosure of Interest: None declared. inherited blistering skin diseases; some forms of the disease are associated with considerable morbidity and increased mortality. The severe forms of the disease are characterized by mutilating AB122 scarring, blisters covering large proportions of the body surface. The Assessment of the Physical Performance, Symptoms, Later in the disease course, mitten deformities, joint contrac- Psychological Status and Quality of Life Before and After tures, oesophageal strictures, corneal erosions, chronic cuta- Hematopoietic Stem Cell Transplantation for Children neous infections, and aggressive squamous cell carcinoma can V. Yıldız1,*,T.Du¨ger1, Z. Avci2,D.Uc¸kan Çetinkaya3 be seen. In this presentation, we presented patient who was 1Health Sciences Faculty, Department of Physical Therapy treated by allogeneic bone marrow and mesenchymal stem cell and Rehabilitation, Hacettepe University, 2Bone Marrow transplantation for epidermolysis bullosa. Transplantation, Dıs¸kapı Yıldırım Beyazıt Hospital, Children Health Materials (or patients) and methods: A 42 day-old male infant and Disease Hematology Oncology, 3Children Hospital Bone was presented to our newborn clinic with vesiculobullous lesions Marrow Transplantation Department, Hacettepe University, on more than his lower legs and forearms including the entire Ankara, Turkey body. The patient diagnosed with dystrophic EB and was performed hematopoietic stem cell transplantation (HSCT) from Introduction: While hematopoietic stem cell transplantation human leukocyte antigen fully matched sibling donor. The bone (HSCT) produces a high cure rate for hematological disease, it is marrow stem cell graft was infused by the intravenous route after also an aggressive therapy that creates physical, social, psycho- the patient was treated with a non-myeloablative conditioning logical and emotional stress for patients. The aim of this study was regimen of cyclophosphamide 50 mg/kg intravenously on day 1, investigate that the effects of HSCT for children on physical rabbit antithymocyte globulin 10 mg/kg intravenously on days 3- performance, symptoms, psychological status and quality of life. 5 and fludarabine 40 mg/m2 intravenously on days 2-5. After Materials (or patients) and methods: Eleven children [girl plasma and red blood cell depletion, the infused nucleated cell n ¼ 3 (%27.3), boy n ¼ 8 (%72.7)], planned to HSCT at Dıs¸kapı and CD34 doses were 13,31 108 cells/kg, 11,27 106 cells/kg, Yıldırım Beyazıt Children Health and Disease Hematology respectively. In addition to the hematopoetic stem cell, Oncology Hospital, were included in this study. They have mesenchymal stem cell derived from the donor (15.9x 106 different hematologic diseases; malignant or non-malignant. nucleated cells/kg) was also transfused by the intravenous route The evaluation of the children was performed before and on the zero day of HSCT. The mesenchymal stem cells were discharge HSCT by the same physiotherapist. Medical records reinfused to the patient on the 13th day of HSCT. We observed were collected: height, weight, body mass index (BMI), type of markedly healing of the skin lesions after the infusions of the transplantation, number of months after diagnosis, occurrence mesenchymal stem cells. Micofenolate mofetil and cyclosporine of complication and GVHD during transplantation. Physical were used for graft versus host disease (GVHD) prophylaxis. performance were assessed by 6 Minute Walk Test (6 MWT), Unfortunately, the patient did not achieve neutrophil, and Time Up and Down Stairs Test (TUDS) and Time Up and Go platelet engraftment; and the chimerism was evaluated 1%, 0% Test (TUG). Muscle strength was assessed by handgrip in 28th day and 49th after transplantation, respectively. GVHD dynamometer, 30 second Sit to Stand Test (SST) and Time prophylaxis was discontinued because chimerism was evaluated Needed to Stand up from Bed Rest Test (TSBT). Pain and 1%. The patient’s skin lesions decreased after transplantation but fatigue level assessed by Wong-Baker Faces Scale, and WeeFIM not completely recovered. The patient’s controls continue in the scale was used as a measure of activity of daily life. Childrens’ hematology outpatient clinic. Depression Inventory (CDI) was used as a measure of Results: The transplantations of hematopetic stem cell with depression level. The PedsQL 3.0 Cancer Module was used MSC at the same time showed beneficial results by clinical as a measure of childrens’ quality of life. And, caregivers’ observation. But non-myeloablative regimen with immunosu- quality of life assessed by Nottingham Health Status (NHS). pression resulted with no-egraftment. Results: Mean age of the children was 6.72±2.96 (min ¼ 3.5, Conclusion: Concominant use of hematopoietic and max ¼ 11) and mean duration after their diagnosis was mesenchymal stem cells transplantation may be a promising 10.90±10.53 months. The results of this study revealed treatment option for patients with epidermolysis bullosa. The decreases of all the parameters that we investigated between non-myeloablative regimen may be a possible reason for graft time points. We found statistically significant decreases in failure. Further studies are necessary to evaluate the optimal TUDS test score (z ¼ -2.524, P ¼ 0.012), TUG test score (z ¼ - HSCT modalities for patients with EB. 2.821, P ¼ 0.005) and SST test score (z ¼ -2.390, P ¼ 0.017). Disclosure of Interest: None declared. Childrens’ quality of life which reported by their parents decreased between time points and it was istatistically AB124 significant (-2.497, P ¼ 0.013). Furthermore, caregivers’ quality of life between time points decreased, at it was statistically Autologous stem cell transplantation for primary bone lymphoma significant (z ¼ -2.395, P ¼ 0.017). X. Tang1,*, Y. Song1,P.De1, Z. Yan1 Conclusion: Physical performance, muscle strength, quality of 1 life of children and parents are affected by aggressive The First Affiliated Hospital of Soochow University, Jiangsu therapies during HSCT. Therefore, there needs to be imple- Institute of Hematology, Suzhou, China mented exercise programmes during HSCT. Disclosure of Interest: None declared. Introduction: Primary bone lymphoma is a rare disease and many of its aspects remain unknown. A number of studies

S606 3. Bacher U,Klyuchnikov E,Carreras J,et al. Conditioning Intensity in Allogeneic Hematopoietic Cell Transplantation(al- loHCT) for Diffuse Large B-Cell Lymphoma(DLBCL).Blood (ASH Annual meting.Abstracts).2011;118.Abstract 501. 4. Fitoussi O, Belhadj K, Mounier N, et al. Survival impact of rituximab combined with ACVBP and upfront consolidation autotransplantation in high-risk diffuse large B-cell lymphoma for GELA.Haematologica,2011,96(8):1136-1143. 5. Ramadan KM, Shenkier T, Sehn LH, Gascoyne RD, Connores JM. A clinicopathological retrospective study of 131 patients with primary bone lymphoma:a population-based study of successively treated cohorts from the British Columbia Cancer Agency. Ann Oncol. 2007;18(1):129–135. Disclosure of Interest: None declared. have been reported from Western countries, but only a few cases were reported from China. The definitions of PBL and the response criteria for PBL were heterogeneous and ambiguous in some of the reports, treatment modalities were also heterogeneous. Combining radiation and multiple courses of Acute leukaemia chemotherapy has been the standard treatment for PBL. However, the roles of hematopoietic stem cell transplantation for PBL remain unclear. Materials (or patients) and methods: In our study, three AB125 patients were admitted to our hospital because of progressive Is the autologous stem cell trasplantation in acute pain in the lesion. The diagnostic of PBL based on PET-CT myelogenous leukemia a good option? Results of a single (bone destruction with abnormal 18 F-FDG uptake) and institution experience immunohistochemical results. Two cases were diagnosed as A. Balerdi1,*, A. Iglesias1, I. Olazabal1, L. Elicegui1, M. Duen˜as1, diffuse large B cell lymphoma (IIE, B group), another was M. Puente1, E. Landeta1, I. Amarika1, E. Amutio1, malignant B cell lymphoma (B cell with T, IV B group). Then J. C. Garcia - Ruiz1 they were treated with 5B6 cycles of chemotherapy with 1Hospital Universitario Cruces, Barakaldo, Spain R-CHOP and achieved complete remission which were confirmed by PET-CT. Then we collected their autologous Introduction: Postremission therapy in acute myelogenous peripheral stem cells (median MNC, 3.64 108/kg; median leukemia (AML) include schedules of dose-intensified cytarabine CD34 þ , 7.45 106/kg) after MAG mobilization chemotherapy. (HDARA C), autologous or allogeneic stem cell trasplantation. Three patients received autologous stem cell transplantation However, the best consolidation treatment remains a topic of with BEAM conditioning regimen. discussion. The last advances in cytogenetic and molecular Results: The median time of neutrophil recovery was 10 days. biology have served to better disease stratification and permit There were no serious transplant related complications. During risk adapted treatment approaches, which have improved the a median follow-up of 2 years, all 3 patients maintain outcome of the disease. Is there a role for autologous stem cell continuous complete remission and go back to work. trasplantation (ASCT) for paients with AML in this situation? We Conclusion: In conclusion, our study demonstrated that present our experience in the treatment of AML with ASCT. chemotherapy with R-CHOP followed by autologous hemato- Materials (or patients) and methods: From 1999 to 2012, 32 poietic stem cell transplantation is safe and effective in adult patients in first complete remission (16 men, 16 women) treating primary bone malignant lymphoma. Follow-up studies received ASCT. The median age was 43 years at diagnosis are necessary to evaluate its long-term effect and to determine (range, 24–66 years). The preparative regimen for ASCT whether it can be used as a first-line therapy. consisted of intravenous busulfan and cyclophosphamide References: 1. Sharathkumar Bhagavathi, MD; Kai Fu, MD, PhD. (BUCY2). For the analysis of the results patients have been Primary Bone Lymphoma.Arch Pathol Lab Med—Vol 133, divided into 3 prognostic subgroups based on karyotype, November 2009. molecular analysis and the need of reinduction to reach 2. Maruyama D, Watanabe T, Beppu Y, et al. Primary bone complete remission. Patients with promyelocityc leukemia lymphoma: a new and detailed characterization of 28 patients have been excluded. in a single-institution study. Japanese journal of clinical Results: In this retropective revision the OS and DFS were oncology,2007,37(3):216-223. estimated by the Kaplan and Meyer method. The overall

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S607 survival (OS) median is 8,895 years and the disease free bone marrow relapse. Showed morphological and molecular survival (DFS) median is 6,790 years. 72,72% of patients in the remission after 2 cycles of chemotherapy with clopharabine unfavourable risk subgroup have relapsed, versus 31,25% and and Ara- C. Allogenic stem cell transplant from matched 40% in intermediate and favourable risk subgroups respec- sibling cord performed. No significant complications after tively. The OS and DFS 5 years is 80% and 60% in favourable transplant. Currently alive in second complete remission 8 group, 56,25% and 68,75% in intermediate group and 54,54% years after diagnosis. Retrospective review of samples at and 36,36% in unfavorable group. The number of consolida- diagnosis showed translocation t(7;12) (q36;p13). tion cycles or the number of HDARA C before the transplanta- Results: See conclusions. tion had no apparent impact on outcome. Conclusion: Although our first patient died in first CR, her final Conclusion: Although the role of ASCT remains controvesial, event was related to transplant related morbidity. Our second in our experience ASCT can provide long term OS and DFS for patient, who shares the same mutation is alive 8 years after patients with AML in first remission and favorable or diagnosis. The t(7;12) and a ectopic expression on HLXB9 is intermediae risk factors. Those with unfavorable karyotype associated with poor outcome in infant AML, therefore an have a high indicence of relapse and might benefite from early diagnosis and intensive treatment including SCT is more aggresive treatments, like allotrasplant. While it is still probably essential for an accurate management. not clear what the best preparative regimen prior ASCT is, we References: High incidence of t (7;12) (q36;p13) in infant AML have used BUCY2 with aceptable results. but not in infant ALL, with a dismal outcome and ectopic Given these results, we think it would be necesary to compare expression of HLXB9. Anne R.M Von Berg et al. Genes, ASCT with HDARA C in randomized prospective trials, focusing chromosomes & cancer 45:731-730 (2006). on patients with favorable and intermediate risk factors. The dual rol of HLXB) in leukemia. Stuart Ferguson et al. Disclosure of Interest: None declared. Pediatric blood and cancer 2011; 56:349-352. Disclosure of Interest: None declared. AB126 t(7;12) (q36;p13): A poor prognosis recurrent translocation AB127 in pediatric AML FLAG-Ida, the possible breakthrough in the treatment of B. Torres Guerola1,*, G. Iacoboni Garcı´a-Calvo2, C. Fuentes refractory acute myeloid leukemia in the context of Socorro1, E. Such Taboada3, M. D. M. Andre´s Moreno1,J.M. hematopoietic stem cell transplantation: Single center Ferna´ndez navarro1 experience 1Pediatric bone marrow transplant, Hospital Infantil La Fe, B. Georgievski1,*, A. Pivkova Veljanovska1, S. Genadieva Stavric1, 2Cytogenetic Laboratory, 3Cytogenetics laboratory, Hospital La Z. Stojanoski1, L. Cadievski1, L. Cevreska1 Fe, Valencia, Spain 1University Clinic for Hematology, Skopje, Macedonia, The Former Yugoslav Republic Of Introduction: The t(7;12) (q36;p13) is a recurrent translocation involving the ETV6/TEL gene (12p13) and a heterogeneous Introduction: Acute myeloid leukemia (AML). The Disease that breakpoint at 7q36. Von Bergh et al showed this translocation we know so much of, but so little that we can actually do. The in 30% of patients with infant acute myeloid leukemia (AML). A modern scientific medicine has entered the micro-universe of fusion transcript between HLXB9 and ETV6 in AML t (7;12) is the malignant cell, describing signaling pathways, but still no ocasionally found. Recently data show poor outcome of magic cure is defined. So, what shall we do with refractory infants with AML and HLXB9/ETV6 rearrangement which is AML, or better to say how to treat patients with this form of the fusion resulting from t (7;12) (q36;p13), fusion transcript AML, giving them chance of better survival? between exon 1 of HLXB9 gene localized in 7q36 and exon 3 Materials (or patients) and methods: Our main inclusion of ETV6. We report the clinical course of 2 patients with AML criteria for evaluation were patients with refractory AML, and with HLXB9/TEL rearrangement. confirmed with reevaluation of bone marrow after standard Materials (or patients) and methods: We have investigated induction therapy (7 þ 3) where we have administered FLAG- t(7;12) (q36;p13)in our pediatric AML series since 2002. Ida regimen (fludarabine 30 mg/m2, AraC 2 g/m2 for 5 days, 80patients evaluated. Out of 80 only two carried the t(7;12) idarubicin 10 mg/m2 for 3 days, and G-CSF 5 micro g/kg from (q36;p13). day 0 until neutrophil recovery) and achieved complete Patient 1:8month old female diagnosed of AML M1. Karyotype remission as preparation for hematopoietic stem cell transplan- with G banding:46,XX, der(7) t(7;12)(q36;p13), t(7;12)(q21;q24) tation (HSCT) as mandatory further treatment of the disease. We [20].The FISH probe of TEL-AML1/HLXB9 was positive in 70% of managed to achieve complete remission in 22 patients with the analyzed nucleus, confirming a t(7;12). The cerebrospinal refractory AML (46%), and all of them underwent HSCT. fluid showed blasts with the same immunophenotype of the Results: We have evaluated 22 patients from the period of leukemia cells. The infant achieved morphological remission 2001 to 2014, 15 males and 7 females. Average age of the after the first block of induction chemotherapy.Bone marrow patients was 36.6 years (17-53). 13 patients received one aspirate (BMA) performed prior to second induction course course of FLAG-Ida, achieved CR and proceeded directly to showed positive MRD 0.5%. She received second course of HSCT, and 9 received two courses, 7 for achieving CR, and 2 as induction and first block of consolidation. BMA post induc- consolidation. We performed autologous HSCT at 13 patients, tion2: MRD 1%. BMA post consolidation 1: morphological and and allogeneic in 9 patients. In the autologous setting, molecular remission. She received second course of consolida- mobilization of peripheral blood stem cells was performed tion and was transplanted. Allogenic unrelated stem cell with etoposide þ G-CSF in 5 patients, and G-CSF only in 8 transplant (SCT) 8/8 performed following conditioning with patients. The average number of collected mononuclear cells busulfan, thiotepa and cyclophosphamide. Major complica- was 2.9 108/KgTT (1.5-6.5). Median time to HSCT was 6.6 tions after SCT including venooclusive disease, skin graft months (4-10). In most of the patients we used myeloablative versus host disease, Cytomegalovirus infection and respiratory conditioning (MAC) with Bu þ Cy (13 patients) and BEAM (8 distress that led to multiorganic failure. She died 11 moths patients). Median time to engraftment was 13.1 days (9-22). No after diagnosis. BMA post SCT showed complete remission significant increase in supportive treatment with blood (CR) with full donor chimerism. products was registered in these patients, and the transplant Patient 2:6 year old male diagnosed of AML M0. Karyotype(G- related mortality was 0. banding): 48XY, þ 8, þ 19 [19]. He achieved complete remis- In the posttransplant period, relapse was confirmed in 9 sion after two cycles of induction. Received 2 cycles of patients. Median time to relapse was 9 months after HSCT. All consolidation and autologus stem cell transplant following patients died despite another attempt with chemotherapy, conditioning with busulfan, cyclophosphamide and etoposide. mainly of infectious complications and heart failure. 13 Two years after the end of his treatment, he presented isolated patients are still alive. The disease free survival (DFS) in our

S608 group of patients is 74 months (22-148). The longest period of 4 patients had graft failure after transplant. Among the others, CR posttransplant is 148 months in patient with allogeneic 23 (31%) resulted in relapse median 30±28 months after HSCT, and registered GvHD. In the autologous setting all transplantation. Isolated extramedullary relapse and bone relapsed patients were mobilized with G-CSF only. The fastest marrow relapse occurred in six and fourteen patients, relapse is in patient transplanted in active disease (7 months). respectively. The 2 –year and 5 –year cumulative incidence Conclusion: We have concluded that FLAG-Ida regimen is of OS was %71±5 and %56±6, respectively. appropriate and suitable salvage chemotherapy protocol for If the patient could not achieve complete remission just before patients with refractory AML especially when it is used in the transplantation, the rate of relapse increased (P ¼ 0.03). Also in context of preparation for HSCT. Performing autologous or the group of patients who underwent transplantation in first allogeneic HSCT in these patients, combined with good remisson, relapse rate decreased than the others (P ¼ 0.04). supportive treatment, may provide prolonged survival rates The following factors did not significantly influence relapse that surely represents a significant benefit for patients with rate after transplantation: type of disease, presence of this form of disease that otherwise is categorized as an entity extramedullary disease or hyperleukocytosis at the time of poor prognostic features. diagnosis, donor type, HLA matching, stem cell source, TBI- Disclosure of Interest: None declared. based regimen. Conclusion: This study showed that remission status at the AB128 time of transplantation is an important factor which influence Relapse of Pediatric Leukemia Following Hematopoietic the rate of relapse after transplant. Stem Cell Transplantation: a Single Center Experience Disclosure of Interest: None declared. B. Akıncı1,*, S. Aksoylar1,S.Go¨zmen1,Z.O¨ nder1, S. Kamer1, S. Kansoy1 AB129 1Ege University,˙ Izmir, Turkey Hematopoietic Stem Cell Transplantation (HSCT) for intermediate-risk Acute Myeloid Leukemia (AML): our Introduction: Hematopoetic stem cell transplantation (HSCT) single center experience is one of the most curative treatment for hematological C. Montes-Gaisan1,*, A. Bermu´dez1,G.Pe´rez1, A. Cuesta1,B.Lo´pez malignancies, helping to achieve long term remission. 1, C. Richard1, E. Conde1 Although significant progress has been made in HSCT, 1Hematology, Hospital Universitario Marque´s de Valdecilla, posttransplant relapse remains one of the most important Santander, Spain cause of treatment failure and major cause of mortality. In this study, we aimed to describe overall incidence of relapse, Introduction: Acute Myeloid Leukemia (AML) requires post- delineate risk factors that may have affected the incidence of remission therapy: consolidations with high dose chemother- relapse and investigate clinical outcomes of leukemia patients apy, intensification with Autologous Stem Cell Transplantation after transplantation. (Auto-SCT) or immune effect after Allogeneic Stem Cell Materials (or patients) and methods: Between February 1999 Transplantation (Allo-SCT). The choice depends on the and June 2013, 78 patients with leukemia underwent HSCT at estimated relapse incidence and the morbimortality expected Ege University Hospital. Informations of these procedures were with the procedure. In this way, low-risk AML can avoid the collected retrospectively from the patients medical records. SCT but high-risk AML needs an Allo-TPH to be cured. Our Data analyses were primarily conducted using SPSS software. objective is to analyze our long-term survival outcome in the The mean age at transplant was 9.9 years (1.3-18.9) and the intermediate-risk AML, which therapeutic options are more ratio of males to females was 1.6:1. Forty-two patients were controversial. transplanted for acute lymphoblastic leukemia and 36 for acute Materials (or patients) and methods: 97 adults, diagnosed myelogenous leukemia. Sixty-nine patients were in first with non-promyelocytic AML and classified as intermediate- complete remission (CR) while 29 patients were beyond 2nd risk according to the NCCN guidelines, underwent SCT in our CR at the time of transplantation. Nine patients underwent center from 1990 to 2010. Auto-SCT was performed in 38 transplant with active or refractory leukemia. patients (median age: 52 years) and Allo-SCT in 59 patients Results: Ninety-two percent of all transplants was allogeneic (median age: 43 years). In the descriptive analysis between and the others were autologous. Donors were matched both groups (Auto-SCT vs Allo-SCT): a) Relapse risk factors: We sibling, matched related, matched unrelated and mismatched found significant differences in etiology (14% secondary vs donor in 47 (65%), 7 (10%), 17 (24%), 1 (1%) patients 29%), extramedullary disease (6% vs 17%), response to respectively. Stem cell source was bone marrow in 30 patients, Induction (5% refractory vs 17%), number of previous peripheral blood in 40 patients and cord blood in 5 patients. treatment lines (11% two or more vs 32%) and pre-SCT Total body irradiation was used as a part of the conditioning disease status (100%RC, 92% in 1st, vs 93%, 81% in 1st). regimen in 20 ( 47%) acute lymphoblastic leukemia patients. However, hyperleucocytosis at diagnosis or aberrant

[AB129]

S609 immunophenotype were well balanced, 20 and 80% respec- tively. b) Transplant related Mortality Risk factors: We found significant differences in year of SCT (81% before 2005 vs 32%), conditioning regimen (75% oral BuCy vs 63% ev BuFlu/ BuCy) and presence of acute Graft-Versus-Host Disease (0% vs 56%, 9% severe forms). Results: Estimated OS at 5 years: 53% in Auto-SCT (median of follow-up: 60 months) and 51% in Allo-SCT (median of follow- up: 39 months). Autologous SCT: 16 patients(42%) relapsed (all of them in the first 2 years post-SCT) and 22 patients(58%) died: 14 because of disease, 3 because of transplant related mortality and 5 because of secondary neoplasms (4 of them were hematologic ones and appeared with a median follow-up of 90 months and without having received radiotherapy before). 16 patients are alived and in CR at the date of the last follow-up, 87% with ECOG 0. Allogeneic SCT: 12 patients(20%) relapsed and 24 patients(41%) died: 12 because of transplant related mortality, 10 because of disease and 2 because other reasons. 35 patients are alived and in CR at the date of the last follow-up, 40% with ECOG Z1. Conclusion: In our series we haven´t found significant differences in OS for intermediate-risk LMA which underwent Autologous or Allogeneic SCT. Auto-SCT seems to be a curative option to be consider in non-fit elderly patients, with an almost inexistent morbimortality in recent years. References: Mro´zek K, Heerema NA, Bloomfield CD. Cytogenetics in acute leukemia. Blood Rev 2004;18: 115-36. Mro´ zek K, Dohner H, Bloomfield CD. Influence of new molecular prognostic markers in patients with karyotypically normal acute myeloid leukemia: recent advances. Curr Opin 375 mg/m2 were administered. The patient’s symptomatic Hematol 2007;14:106-14. anemia persisted and she continued to receive bi-weekly Disclosure of Interest: None declared. RBC transfusions until pancytopenia was noted on CBC 6 months after HSCT. Bone marrow examination showed the AB130 presence of %30 myeloblasts. Flow-cytometric analysis revealed Acute myeloid leukemia of donor cell origin after that the blasts were positive for CD13, CD33, CD11b and CD15. allogeneic stem cell transplantation in a patient with acute Interphase FISH carried out on bone marrrow aspirate by using myeloid leukemia the CEP X,Y. DNA probe kit (Abbott Vysis) showed an XY signal pattern in 196/200 interphase nuclei cells indicating sustained F. Hindilerden1,*, B. Kantarcioglu2, S. Guvenc1, H. S. Goksoy1,M. 3 4 1 4 full donor chimerism (Image). Acute myeloid leukemia of donor Buyukdogan , T. Ozcelık , R. Diz Kucukkaya , M. Arat origin was diagnosed. Remission reinduction chemotherapy 1Istanbul Bilim University, Department of Internal Medicine Division 2 including mitoxantrone, etoposide and cytarabine was initiated. of Hematology, Istanbul Okmeydanı Training and Research Patient succumbed to Klebsiella septicaemia on the 19th day of Hospital Department of Internal Medicine Division of Hematology, remission reinduction. 3Gayrettepe Florence Nightingale Hospital Department of Medical 4 Conclusion: In an EBMT survey covering 10489 HSCT patients Genetics, S¸is¸li Florence Nightingale Hospital Adult Hematopoietic between 1982 and 2003, 14 cases of acute donor cell leukemia Stem Cell Transplantation Unit, Istanbul, Turkey were identified. In sex mis-matched bone marrow transplant recipients, it is simple to diagnose this very rare condition by Introduction: Acute leukemia relapse after HSCT often occurs demostration of the opposite sex in the leukemic cells by due to return of an original disease clone. Yet, rarely acute conventional cytogenetics or FI˙SH. Further studies regarding leukemia can develop de novo in engrafted cells of donor origin. the incidence, potential pathogenic factors, therapetic options Materials (or patients) and methods: A 51-year-old woman st and outcome of patients from donor cell leukemia are needed diagnosed with acute myeloid leukemia obtained 1 hema- to a beter understanding of this rare entity. tological remission following 1 course of 7 þ 3 and 1 course of Disclosure of Interest: None declared. FLAG-I˙DA. She underwent myeloablative peripheral allogeneic HCST from her HLA-identical and mismatched major blood group mismatched brother. BU/CY was the conditioning AB131 regimen and cyclosporine and methothrexate were used for CD34-enriched, T-cell repleted PBSCTs from unrelated GVHD prophylaxis. The donor was healthy at time of mismatch donors as alternative stem cell source in high- peripheral stemcell harvest with normal CBCs. Thrombocyte risk pediatric acute lymphoblastic leukemia: A single and leukocyte engraftment were achieved on day (D) þ 11 center experience and D þ 16, respectively. J.-S. Ku¨hl1,*, S. Voigt1, G. Strau1, C. Eckert1, A. von Stackelberg1, Results: On D þ 30 after HCST, full donor chimerism (46 XY) was W. Ebell1 shown by FI˙SH. RBC transfusion was required for symptomatic 1Pediatric Oncology/Hematology/SCT, CHARITE BERLIN, Berlin, anemia which continued in the presence of high titers of anti Germany red blood cell antibodies. Five courses of plasmapheresis were performed. Yet, symptomatic anemia persisted 3 months after Introduction: For children with high-risk ALL lacking a HSCT. Serology was negative for parvovirus and CMV DNA matched donor, the optimal alternative donor and stem cell remained negative. Bone marrow biospy on D þ 100 showed source, i.e. haploidentical related vs. cord blood vs. mismatch hematological remission and marked supression of erhytropoi- unrelated, has yet to be determined. We retrospectively esis. These findings were compatible with pure red cell analyzed patients with pediatric ALL who were transplanted aplasia. Full donor chimerism (46 XY) was reconfirmed from a mismatch unrelated donor in comparison to ALL MUD- by FI˙SH. CSa was stopped and 4 weekly courses of Rituximab transplants within the ALL SZT-BFM 2003 trial.

S610 Materials (or patients) and methods: From 2004-2013, a total Results: Mucositis was the dominant manifestation of regi- of 16 ALL pts. (age: 2-21 years) underwent MMUD-PBSCT (HLA- men-related toxicity (RRT), however, grade4II was noted in 08/10-identical: n ¼ 14, HLAo08/10-identical: n ¼ 2). ALL-type: 13 only three cases (20%). Other observed forms of organ RRT did precursor B-cell (including 2 proB), 2 T-lineage, 1 biphenotypic not exceed grade I. The engraftment of hematopoiesis with leukemia; 4 pts. were in 4CR2, 1 pt. in partial 2nd remission (PR2). complete donor chimerism was achieved in all patients. In 13 12 pts. received the former ALL SZT-BFM 2003 protocols for patients (87%) dose reduction and full withdrawal of CyA were MMDs (TBI/Flu/VP16/ATG-F in N ¼ 10). PBSCs were CD34 þ within a median of 46 (range, 28-77) and 113 (range, 92-195) selected by immune magnetic sorting using the CliniMacss days after SCT, respectively. Acute GVHD was observed in two device resulting in median CD34 þ cell numbers of 9.0x10E6/kg cases (13%); the chronic form was not noted. Two patients (range 1.8-22.9) followed by a T-cell repletion with 1.0x10E6/kg (13%) relapsed with ALL at 3 and 16 months after CD3 þ cells (range 0.6-1.8) except one. GVHD prophylaxis transplantation. During the above follow-up, all 15 recipients consisted of ATG-Fresenius (N ¼ 15) plus cyclosporine A. were alive, with a probability of 2-year disease-free survival of Results: 9 pts. are alive and well with an estimated 5- yr-OS of 79% (95% CI 54-100%). 62.5±12.1% (minimum follow-up 16 months). Death resulted Conclusion: Although the results were obtained with a small from relapses (N ¼ 2) or TRM (N ¼ 5: 1 cGVHD, 1 neurotoxicity, pilot study group it may be assumed that such approach may be 3 infections). Engraftment was 100% with a median time for considered as an alternative to SCT after traditional myeloablative ANC 4500/ml of 17 days. A boost was given in 2 pts. for initial or reduced conditionings with standard GVHD prophylaxis. poor marrow function. Acute GVHD II was observed in 3 pts. Supported by the project LF-2014-001 of Palacky University in with no aGVHD 4II. However, one aGVHD progressed to an Olomouc. extensive cGVHD resulting in death. In comparison to own ALL Disclosure of Interest: None declared. MUD transplants, there were no significant differences observed with respect to OS (MMUD 56% vs. MUD 74%), AB133 TRM (31% vs. 14%), relapses in CR (7% vs. 20%), and aGVHD Z Assessing the efficacy of allogeneic hematopoietic stem II (19% vs. 32%), respectively. cells transplantation (allo-HSCT) by analyzing survival end Conclusion: As far as low numbers allow to conclude, CD34- points in defined groups of acute myeloid leukemia enriched, T-cell repleted PBSCs from HLA-8/10- or even 07/10- patients; a retrospective, multicenter PALG study identical unrelated donors offer a feasible transplant option for 1,* 2 3 4 high-risk ALL in children lacking a matched donor. Moreover, S. Grosicki , J. Holowiecki , K. Kuliczkowski , A. B. Skotnicki , A. Hellmann5, K. Sulek6, A. Dmoszynska7, J. Kloczko8, OS and relapse rates seem to compare favourably to published 9 10 11 data from haploidentical related transplants. Therefore, W. W. Jedrzejczak , B. Zdziarska , K. Warzocha , A. Wierzbowska12, M. Komarnicki13, A. Pluta12, S. Giebel2 manipulated grafts from unrelated donors with limited 1 mismatches may be prospectively evaluated in comparison Department of Hematology, SPZOZ ZSM w Chorzowie, Chorzow, 2Department of BMT, Comprehensive Cancer Center, Gliwice, to other alternative stem cell sources. 3 Disclosure of Interest: None declared. Department of Hematology, Medical University, Wroclaw, 4Department of Hematology, Jagiellonian University, Cracow, 5Department of Hematology, Medical University, Gdansk, AB132 6Department of Hematology, WIM, Warsaw, 7Samodzielna Allografting after reduced intensity conditioning with Pracownia Transplantologii Klinicznej, Medical University, Lublin, thymoglobulin followed by early withdrawal of 8Department of Hematology, Medical University, Bialystok, prophylactic immunosuppression could be an effective 9Department of Hematology, Medical University, Warsaw, approach to the patients with acute lymphoblastic 10Department of Hematology, Medical University, Szczecin, leukemia 11Department of Hematology, Institute of Hematology and L. Raida1,*, Z. Rusinakova1, R. Szotkowska1, A. Kuba1, E. Faber1, Blood Transfusion, Warsaw, 12Department of Hematology, P. Rohon1, S. Tomas1, K. Indrak1, M. Jarosova1, M. Divoka1, Medical University, Lodz, 13Department of Hematology, Medical Z. Pikalova1, I. Skoumalova1, T. Papajik1 University, Poznan, Poland 1Dpt. of Hemato-Oncology, University Hospital and Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Introduction: The importance of allo-HSCT treatment for Republic survival outcomes in AML patients currently remains unclear. The study aimed to compare measures of clinical treatment for Introduction: Allogeneic stem cell transplantation (SCT) is an AML patients in CR1 (the first complete remission) with or important treatment modality in patients with acute lympho- without being subjected to allo-HSCT. These consisted of blastic leukemia (ALL) and reduced intensity conditionings leukemia-free survival (LFS), overall survival (OS), cumulative (RICs) can be used in the significant proportion of them. incidence of relapse (CIR) and non-relapse mortality disease However, lower cytoreductive potential of RIC results in (NRM). insufficient suppression of the residual leukemic clone. In Materials (or patients) and methods: Subjects were 622 case of standard graft-versus-host disease (GVHD) prophylaxis, patients, median age of 44, forming part of the prospective, graft-versus-leukemia (GVL) reaction may be suppressed for a randomized and multicenter clinical PALG trials during 1999- relatively long time and in combination with the lower 2008. The Mantel-Byar approach was used to assess allo-HSCT reduction of the residual leukemic clone following RIC, this on survival endpoints, accounting for a changing transplant may lead to a higher post-transplant recurrence of ALL. status. Materials (or patients) and methods: 15 patients with high- Results: Undergoing allo-HSCT significantly improved the LFS risk ALL in complete remission and a median age of 34 (range, and OS for the entire group of AML patients in CR1, along with 22-58) years underwent allogeneic SCT after RIC combining the DAC induction subgroup and for the group with fludarabine (150 mg/m2) and melphalan (140 mg/m2) with unfavorable cytogenetics aged 41-60. thymoglobulin (4.5 mg/kg). 8 of them (53%) were allografted The CIR demonstrated, that allo-HSCT reduced the risk of from HLA mismatched donors and mycophenolate mofetil was relapse for AML patients in CR1 and those with an unfavorable added to post-transplant GVHD prophylaxis with cyclosporin A cytogenetic risk. In addition, the NRM analysis showed that, (CyA) in these recipients. ‘‘In vivo’’ T-cell depletion reducing the allo-HSCT significantly reduced the risk of death unrelated to risk of GVHD was compensated by early initiation of reduction relapse for the entire group of AML patients in CR1 aged 41-60. (planned on days þ 28-35) and withdrawal (planned on days Conclusion: The allo-HSCT treatment particularly benefitted þ 90-110) of prophylactic immunosuppression aimed at survival for the AML in CR1 group having an unfavorable maintaining GVL effect. The median post-transplant follow- cytogenetic prognosis. up was 19 months (range, 7-67 months). Disclosure of Interest: None declared.

S611 AB134 outcome of children with refractory or relapsed leukemia after Factors determining outcome of hematopoietic cell allo-HSCT. transplantation in patients with acute myeloid leukemia at Materials (or patients) and methods: We retrospectively King Faisal Specialist Hospital and Research Center, reviewed the medical records of 21 children (three girls and 18 Riyadh, Saudi Arabia boys) who underwent allo-HSCT for refractory or relapsed S. Al-Sweedan1,*, A. Belgaumi1, M. Ayas1, A. AL-AHMARI1, acute lymphoblastic leukemia (ALL) (n:11) or acute myelo- K. Alkayed1, H. El-Solh1, A. Al-Jefri 1, K. Siddiqui1, R. Jafri1, blastic leukemia (AML) (n:10). M. Viqaruddin 1, N. Farhan 1, A. Mohamed1, A. Al-Seraihi 1 Results: The median age of 21 children was 7 years (range: 1PEDIATRIC HEMATOLOGY/ONCOLOGY/BMT, KFSH&RC, Riyadh, 2-15 years) at the time of transplantation. The donor Saudi Arabia was an HLA-matched related siblings in 15 children and haploidentical mother or father in six children. Second Introduction: Allogeneic hematopoietic cell transplantation complete remission (CR2) prior to transplantation was has significantly improved outcome for patients with Acute obtained in 11 children (8 ALL, 3 AML) while 10 children Myeloid Leukemia (AML), in particular in those who are in first patients (3 ALL, 7 AML) were unresponsive to salvage complete remission (CR1) and in recipients of matched related chemotherapy. Subsequently, clofarabine or fludarabine donors. containing cytoreductive therapy followed by myeloablative Materials (or patients) and methods: Medical records of 73 regimen for refractory children and standart myeloablative patients with AML who underwent HCT at our institution regimen for CR2 children were given. The source of stem cells was peripheral blood in 14 of 21 children. Stem cell grafts between 2005 and 2011were reviewed after getting approval 6 6 of the project from our institution review board. Data contained a median of 5,6x10 /kg (range 1.8-29.9x10 /kg) extracted included those related to clinical characteristics of CD34 þ cells/kg infused children. Myeloid engraftment at a the patients and their outcome. Outcome metrics included median of 13 days (range: 10-24 days) and platelet overall (OS) and event free survival (EFS), cumulative incidence engraftment at a median of 16 days (range: 10-36 days) were of GVHD, count recovery and post-transplant complications. noted. All children had complete chimerism at day 28 Results: Forty six patients were male (63.0%). Median age at and this has persisted until 3 months. There were no HCT was 7.9 years (range 0.64- 14.22), median time to HCT transplant related mortality. Eleven children developed grade after diagnosis was 6.5 months. Eight patients were below the 2-3 skin, liver or intestinal GVHD. Of the 21 children, 15 (7 ALL, age of one year (11%). All patients were in CR at the time of 8 AML) relapsed at a median of 6,5 months (range:3,1-47,5 HCT, 54 patients were in CR1, 18 in CR2 and 1 in CR3 at the months) after allo-HSCT while six are still alive in CR2 for a time of HCT; 51 patients received HCT from HLA-identical median 18,7 months (range:4.1-22.5 months). The relapsed related donors and 22 from other donors (1 or 2 antigens rate in CR2 children (90%) was significantly higher than in mismatch cord blood (CB) or one antigen mismatch sibling or refractory children (50%) (p:0.04). Children who relapsed after related bone marrow(BM)). Stem cell source was BM in 55 HSCT were treated with intensive chemotherapy combined (75.3%), CB in 17(23.28%) and peripheral blood in 1(1.36%). All with donor lymphocyte infusion (DLI) (n:8), only chemotherapy but one patient received myeloablative conditioning. The OS (n:4) or second allo-HSCT (n:3). Of these 15 relapsed children, in our group of patients was 51.8% and the EFS was 48%. three children with CR2 (30%) and one refractory child (20%) Median follow-up time for the cohort was 50.667±2.4 months were alive with intensive chemotherapy þ DLI (n:2) or second (95%CI: 45.9-55.4). 39 patients were alive with a median allo-HSCT (n:2). follow-up time of 50.1 months (Min: 1.8, Max: 111.8). 16 After a median follow up of 15 months from allo-HSCT, 10 of 21 patients survived for more than 5 years (Min: 65.2, Max: 111.8) children (5 ALL, 5 AML) were alive. At 2 years, the probabilities of months. The cumulative incidence of acute GVHD was 6.8±2.9 overall survival and event free survival (EFS) and the relapsed and of chronic GVHD was 9.9±3.6. Median time to ANC and rate in the entire group were 41%, 35% and 64%, respectively. platelet recovery was 16 days (range 9-37) and 29 days (range Conclusion: Our data shows that cytoreductive therapy 15-180) respectively. Three patients acquired CMV infection followed by allo-HSCT can induce sustained complete remis- after transplant. No one developed VOD, Hemorrhagic cystitis sion in about a half of the children with refractory leukemia. or other major complications. There was a significant impact of Intensive chemotherapy plus DLI or re-transplantation may be patient’s age at diagnosis on the overall survival (Infantile: considered in post-tranplant relapsing cases. 100%, others: 45.6%±6.4%, P ¼ 0.016) and event free survival Disclosure of Interest: None declared. (Infantile: 100%, others: 40.6%±7.1%, P ¼ 0.013). Percentage of blasts at transplant, patients or donors gender, donor source and HLA disparity did not significantly affect OS or EFS. Conclusion: Our results show a favorable outcome to HCT for AML patients comparable to published data. Our analysis also Aplastic anaemia showed that infantile leukemia was associated with superior outcome. Disclosure of Interest: None declared. AB136 Successful liver transplantation for Wilson’s disease as a AB135 rare cause of hepatic failure after SCT in a child with Allogeneic stem cell transplantation in children with Fanconi aplastic anemia relapsed and refractory leukemia C. Albayrak1,*, D. Albayrak1, A. G. Kalaycı2, A. Bıc¸akcı3, M. Elli4 Z. Kaya1,*, M. Karaku¨kc¸u¨2,E.U¨nal2,U¨. Koc¸ak1, I. Yenicesu1, 1Pediatric Hematology Department, 2Pediatric Gastroenterology, T. Gu¨rsel1 Ondokuz Mayıs University Medical Faculty, Samsun, 3General 1Pediatric Bone Marrow Transplantation Divisions of the Surgery,˙ Ino¨nu¨ University, Malatya, 4Pediatric Oncology, Ondokuz Pediatric Hematology Departments, Medical School of Gazi Mayıs University Medical Faculty, Samsun, Turkey University, Ankara, 2Pediatric Bone Marrow Transplantation Divisions of the Pediatric Hematology Departments, Medical Introduction: After allogeneic stem cell transplantation liver School of Erciyes University, Kayseri, Turkey toxicity may be due to several reasons, such as drug toxicity, graft versus host disease, hepatic veno-oclusive disease. Introduction: Survival rates of children following allogeneic However, other rare causes should be in mind. We present a hematopoetic stem cell transplantation (allo-HSCT) for acute child developing liver failure after allo-SCT and then diagnosed leukemia have markedly increased over the last decades, but Wilson’s disease. The patient was successfully treated with refractory or relapsed leukemia remain the major cause of repeated plasmapheresis for acute hepatic failure and then treatment failure. The aim of this study was to evaluate the transplanted cadaveric liver.

S612 Materials (or patients) and methods: 13-year-old male patient was diagnosed as Fanconi aplastic anemia and stem cell transplantation was performed from HLA full matched sister. Reduced conditioning regimen was consisted of fludarabine plus cyclophosphamide. Liver enzymes and bilirubin levels were moderately elevated after engraftment. Hepatitis markers were negative. We thought the hepatic toxicity to be due to conditioning regimen. Abdominal acid did not developed. Total bilirubin levels were 10-15 mg / dl during the first three months and remained relatively stable around. Direct bilirubin levels began to rise in the fourth month of the patient, coagulation tests were became abnormal. Percutaneous liver biopsy was taken. In the preliminary report of biopsy, toxic hepatitis was considered. All drugs including cyclosporine A were stopped except trimethoprim sulphamethoxazole. Bilirubin values rose during this period. Total bilirubin was 35.7 mg/dL, and direct bilirubin 26.7 mg/dL, ammonia 194.6 mcg/dl, and ceruloplasmin 47.1 mg/dl. Patients were considered going to hepatic coma and started to receive therapeutic plasmapheresis. The total seven times plasmapheresis were made. The first five plasmapheresis were made with fresh frozen plasma. Allergic infection (n ¼ 5), infection (n ¼ 3, incl. ADV-pneumonia), aGvHD reactions occurred in the last plasmapheresis. One week later, and cGvHD (n ¼ 2) and tongue cancer (n ¼ 1). two plasmapheresis were made with albumin. Conclusion: Our results suggest that HSCT for pts. with FA is a Results: Liver biopsy was reported as toxic hepatitis and valid therapeutic option. HSCT characterized by rather high copper accumulation. The copper level of 24- hour urine was incidence of GVHD and infections. Second HSCT is associated high levels. The patient was diagnosed as Wilson’s disease. with high risk of refractory course of infection and GvHD. D-penicillamine and zinc therapy was started. Cadaveric liver Cancer is a cause of late mortality. transplantation was performed one month later. Two years Disclosure of Interest: None declared. after stem cell transplantation the patient is disease free, with complete donor chimerism in bone marrow and stable hepatic AB138 graft function with immunosuppressive therapy. Peripheral Blood Stem Cell Transplantation From Conclusion: As a result, in hepatic toxicity after allo-SCT, Matched Sibling Donor In Children With Severe Aplastic Wilson’s disease should be in mind. The patients with Anemia – Single Center Experience autosomal recessive diseases such as Fanconi aplastic anemia 1,2,* 1,2 2,3 2,3 2,3 may be have another autosomal recessive disease. For this M. E. I. W ,A.G.AN ,A.G.AA ,S.NA ,E.SA , M. N M2,3,M.S1,2 reason, a detailed family medical history should be taken. 1 2 3 Disclosure of Interest: None declared. Pediatrics, Faculty of Medicine, Ain Shams University, Clinical Pathology, Faculty of Medicine, Cairo, Egypt

AB137 Introduction: Aplastic Anemia (AA) is a life threatening Long-time follow-up of patients with Fanconi anemia: two hematological disease. Cure can be achieved in a large centers experience proportion of patients with prompt appropriate treatment. E. Skorobogatova1,*, D. Balashov2, K. Kirgizov1,2, Y. Skvortsova2, According to protocol of Pediatric Hematology Team of BMT M. Persiantseva2, P. Trakhtman2, G. Novichkova2, Unit, Ain Shams University Hospital, children diagnosed with A. Roumyantsev2, A. Maschan2 severe and very severe AA with matched sibling donors proceed 1The Russian Children’s Research Hospital, 2Dmitry Rogachev to peripheral blood stem cell transplantation (PBSCT) as first line Federal Research Center of Pediatric Hematology, Oncology and therapy, whereas children without a donor receive IST. Immunology, Moscow, Russian Federation Materials (or patients) and methods: We report the outcome of 10 children with severe aplastic anemia (AA) who received Introduction: Fanconi anemia (FA) is a rare disease requiring allogeneic transplants from fully matched sibling donors HSCT. Long-time follow-up is very important for future prognosis. (MSD) between April 2011 and June 2014. The median Materials (or patients) and methods: Twenty nine children patients’ age was 7 years with a range of (3.5-13 y). Six were with FA (11 males, 18 females), median age 9,5 (3,7-15,4) years, females and 4 males. All were transplanted using a condition- received HSCT from 17 (58,6%) matched siblings and 12 ing regimen of Cytoxan 200 mg /kg and ATG Fresenius 60 mg/ (41,4%) MUD between November 1994 and April 2014. Graft kg. GVHD prophylaxis included cyclosporine and a short sources: BM (n ¼ 18), PBSC (n ¼ 7), BM þ UCB (n ¼ 3), UCB course of methotrexate (MTX). Median time for patients to (n ¼ 1). One sibling HSCT was 9/10, two MUD were 10/10. Two pass to transplant was 6 months with range of 3 -18 months. pts. were transplanted after secondary MDS development. All patients received PBSCT in a dose of 7 106 CD34 þ cells/ Conditioning regimen: MUD – Bu 4 mg/kg, Cph 20 mg/kg, Flu kg bodyweight, collected after donor mobilization with G-CSF 150 mg/sq.m., ATG; MRD – Bu 4 mg/kg, Flu 150 mg/sq.m., ATG. 15 mg/kg for 4-5 days. GvHD prophylaxis was different – CsA based for MRD HSCT Results: Neutrophil engraftment occurred after a median time and Tacro based for MUD HSCT. of 13 days, with a range of 12-18 days.The median time for Results: All patients engrafted. Rejection rate was 13,8% (n ¼ 4) platelet engraftment was 17 days with a range of 16-26 days. (on 1, 2, 6, 12 months after HSCT). Second HSCT was performed The longer duration to platelet engraftment was observed in 2 in all cases. 2 patients survived after the second HSCT with good patients with major and bidirectional ABO blood group engraftment, 2 patients died after 2nd HSCT (causes of death – incompatibility. Complete donor chimerism (498%) was GvHD, ADV-pneumonia). Direct toxicity of conditioning regimen achieved in all patients. Acute GVHD grade 1, limited to skin, was minimal. No signs of GvHD were observed in 16 pts. (55,2%), occurred in half of patients and responded to a short course of grade I-II aGVHD - 8 pts. (27,6%), grade III-IV – 5 pts. (17,2%). methylprednisone. Chronic GVHD (ocular) occurred in one Limited cGvHD was diagnosed in 3 pts., extensive cGvHD – in 2 patient 9 month post transplant, while starting gradual pts. Median follow up - 31,9 months (3,8-246) the estimated withdrawal of cyclosporine. The median duration of follow probability of 5 years OS was 67,4%, EFS – 62,7%. Two pts. up was 2 years with a range of 1–4.5 years. Early CMV developed oral cavity cancer. The causes of death were GvHD þ activation (day þ 8) occurred in one patient and day( þ 35) in

S613 another patient, both responded well to gancyclovir therapy. leukemogenesis. Hypotheses are generated regarding various Two patients died due to acute toxicity in the form of influences on the pathogenesis of leukemia, such as the transfer disseminated adenovirus infection (day þ 10), and intracranial of dormant oncogenes, stimulation of a receptive donor-cell hemorrhage associated with cardiopulmonary arrest (day clone, or the transmission of viruses. In addition, the host þ 12). Eight patients are currently alive with near normal microenvironment interacting with donor hematopoietic stem quality of life. The overall survival was 80%. Early immune cells may also play a role. This aspect is highlighted in the reconstitution studies using flowcytometry for detection of following case where an identical B cell clone was found in CD19, CD16, CD3, CD4, CD8and CD59, done at 3 and 6 months donor and recipient but the temporal occurrence of the Diseuse revealed satisfactory results. and its precursor stages seemed to be affected in a different Conclusion: We conclude that MSD-PBSCT can cure a large manner. number of patients with severe AA when performed at an Materials (or patients) and methods: We report on a female uncomplicated disease course, even with a delay of 46months. patient (pt) who was diagnosed with a secondary acute myeloid Cytoxan/ATG is a good conditioning regimen, associated with leukemia (AML) 10/2001 at the age of 57 years. After remission very low acute and chronic GVHD- even with the use of PBSC induction an unrelated allogeneic stem cell transplantation grafts- and with satisfactory early immune reconstitution. (allo-SCT) after reduced intensity conditioning was performed in Disclosure of Interest: None declared. 05/2002. The 47-year-old male donor was HLA-identical (10/10). Prophylaxis against graft versus host disease consisted of AB139 cyclosporine which was stopped on day 147. Diamond-Blackfan anemia VS acquired pure red cell Results: Eleven years after transplantation (06/2013), the aplasia: clinical and RPS19 gene mutation patient complained of weight loss due to acute gastritis with fatigue and back pain. No other B symptoms were noted. The S. Jiang1,*, H. jiang2 1 2 clinical examination was unremarkable. The immunophenotyp- Department of hematology, Shanghai Children’s Hospital, ing (IP) of the bone marrow (BM) and the peripheral blood (PB) Shanghai Jiao Tong University, shanghai, China showed the presence of a monoclonal B-lymphocyte popula- tion (MBLP) of 2.61 Gpt/l with the IP CD19 þ CD20 þ CD5 þ Introduction: o investigate clinical similarities and differences, CD10 þ /-CD23 þ CD38 þ (het)FMC7-sIg þ (lambda). The chi- as well as ribosomal protein S19 (RPS19) gene mutations merism by STR analysis remained at 100%. The diagnostic between patients with acquired pure red cell aplasia (PRCA) work-up revealed an unbalanced rearrangement between the and congenital PRCA (Diamond-Blackfan anemia, DBA). long arms of chromosome 4 and 17 in the karyotype analysis in Materials (or patients) and methods: 20 children with PRCA 14 of 20 analyzed metaphases. Due to the sex chromosome from multiple centers were enrolled, including 14 DBA and 6 constellation, the cells of the aberrant clone could be identified acquired PRCA. The age of disease onset, peripheral blood counts, as donor cells (45, XY,der(4)t(7;17)(q35;q11),-17). In FISH, the and responses to treatment were compared between these two TP53 deletion (17p deletion) was confirmed. Three months later groups. The RPS19 gene sequences of these children were (11/2013), further features of overt lymphoproliferative neopla- determined using the Sanger dideoxy chain termination method. sia were still absent but IP confirmed the presence of MBLP with Results: A total of 42.8% of the DBA patients exhibited 3.6 Gpt/l in the PB. In summary, a donor-related monoclonal combined congenital malformation. Compared with the B-lymphocytosis (MBL) was diagnosed. In 09/2014 the patient acquired PRCA patients, DBA patients displayed an early age had no features of chronic lymphocytic leukemia (CLL). The IP of disease onset (Po0.05) and a lower number of reticulocytes still showed a MBL population of 3.8 Gpt/l. After a request to the (Po0.05), whereas their peripheral white blood cell (WBC), donor registry the transplant center was informed that the hemoglobin (HGB), and platelet (PLT) counts did not show any former unrelated stem cell donor has developed B-CLL (Binet difference. The efficacy of corticosteroids treatment for DBA stage A) in 11/2010, 8 years after blood stem cell donation. This was 78.6%, while that for acquired PRCA was 100% (P40.05). CLL had the same IP as the MBL of the recipient and TP53 RPS19 point mutations were detected in 7.1% of the DBA deletion was also localized to chromosome band 17p13. In patients, but not in any of the acquired PRCA patients. addition, the recipient’s IGHV status showed an IgHV3-7 * 02 Conclusion: Patients with DBA have lower reticulocyte counts mutation in 8% and was identical with the IGHV mutation status than acquired PRCA patients. Unlike DBA, acquired PRCA of the donor. During the time course the former stem cell donor might not be associated with RPS19 abnormality. was in need of treatment after progression of the B-CLL. Finally, References: 10. Danilova N, Sakamoto KM, Lin S. Ribosomal allo-SCT was performed in 04/2014 in the donor. In contrast, 15 protein S19 deficiency in zebra fish leads to developmental months after the diagnosis of MBL there is still no transforma- abnormalities and defective erythropoiesis through activation tion into B-CLL detectable in our patient (recipient). of p53 protein family. Blood. 2008; 112(13): 5228-5237. Conclusion: This is the first report of proven donor-derived Disclosure of Interest: None declared. MBL in a recipient after allo-SCT, whereas the donor developed clinically relevant B-CLL years before. The case suggests that the host micro-environment may have influenced the various patterns of disease progression despite of the identical clone Chronic leukaemia of origin. Disclosure of Interest: None declared.

AB140 AB141 Donor-derived monoclonal B-lymphocytosis after Outcome of allogeneic stem cell transplantation for allogeneic peripheral blood stem cell transplantation patients with primary myelofibrosis: a single-center A. Klink1,*, J. Fischer2, V. Schmidt1, H. G. Sayer1,3, I. Hilgendorf1, experience A. Hochhaus1, M. von Lilienfeld-Toal1 F. Vural1,*, N. Soyer1, F. Celik2, M. Comert1, F. Sahin1, G. Saydam1, 1Department of Internal Medicine II, Hematology and Oncology, M. Tombuloglu1 Jena University Hospital, Jena, 2University Hospital, Institute for 1Hematology, 2Internal Medicine, Ege University,˙ Izmir, Turkey Transplantation Diagnostics and Cell Therapeutics, Heinrich- Heine-University Du¨sseldorf, Medical Faculty, Duesseldorf, Introduction: The only known curative therapy for primary 3Department of Internal Medicine IV, Hematology and Oncology, myelofibrosis (PMF) is allogeneic hematopoietic stem cell Helios Klinikum, Erfurt, Germany transplantation (allo-HSCT). Although reduced intensity conditioning (RIC) allo-HSCT has been increasingly used to Introduction: Donor cell derived leukemia is a rare complica- treat elderly patient, myeloablative conditioning has been tion but it may give interesting insights into in-vivo generally used to treat younger patient.

S614 Table 1. Clinical characteristics and follow up course of our PMF patients

Case no DIPSS skor Conditioning Acute GVHD Onset Chronic Outcome Relapse regimen Grade Localization GVHD type Current status Cause of death

1 High Cy-Bu -70 -2 - - -- Alive - 2 High Melphalan Liver Limited 73 Alive - 3* High Cy-Bu -- - - 1151 Died on day 749 Relapse 4* High Flu-bu-ATG - - Died on day 542 Relapse 5 High Flu-Ara-C- Cy - - - 116 Died on day 258 relapse 6 High Cy-Bu - - - - Alive - 7 High Cy-Bu 12 2 Liver Limited - Alive - 8 High Cy-Bu - - - - Alive - 9 High Cy-Bu - - - - Alive - 10 High Cy-Bu 40 4 Skin - - Alive - 11 High Cy-Bu 12 4 and 3 Gut and Liver - - Died on day 68 GVHD

Materials (or patients) and methods: A total of 10 patients (5 types as primary and secondary HLH. All the types are male and 5 female) with PMF underwent 11 alloHSCTs characterized by clinical signs and symptoms of extreme between 2005 and 2014. We evaluated the clinical character- inflammation. Replacing the patient’s immune system with istics and follow up course of our patients who undergone that of a healthy donor by hematopoetic stem cell transplan- allo-HSCT at a single transplant center retrospectively. tation (HSCT) is the only curative option for patients with Results: The median age of evaluated patients was 60.5 years (22- primary HLH as well as for patients with secondary HLH who 62 years). The median time from diagnosis to transplantation was fail to respond to therapy. Long-term survival reports of 31 months (4- 53 months). Transplanted stem cell sources were 1 HSCT varies from 50% to 70%, depending on the type of from unrelated and 10 from related donors. Myeloablative donor and the intensity of the conditioning regimen. conditioning regimen consist of intravenous busulfan at Transplant related mortality (TRM) is the major problem for 12.8 mg/kg and cyclophosphamide at 120 mg/kg was applied to HSCT. 8 patients and RIC regimen (fludarabine or melphalan containing In this study, we report the results of HSCT in 16 consecutive regimens) applied to 3 patients. Methotrexate and cyclosporine patients with HLH transplanted at our center, Akdeniz were used for graft-versus-host disease (GVHD) prophylaxis. The University Pediatric Hematopoietic Stem Cell Transplantation median number of infused CD34 þ cells was 6.8 (3.2-10.4) 106 Unit. cells/kg. Neutrophil and thrombocyte engraftment were achieved Materials (or patients) and methods: Between September in median þ 22 days (12- 31 days) and þ 19.5 days (13- 56 days) 2006 and December 2014, 16 patients underwent HSCT from respectively.AcuteGVHDwasseen4of10patientsand2ofthem either a related (fully matched related donor [MRD], n ¼ 3) or were grade 3- 4 GVHD. Chronic GVHD was seen 2 of 10 patients. an unrelated donor (fully matched unrelated donor [MUD], Two of 10 patients had relapsed after transplantation and both of n ¼ 3, one antigen-mismatched unrelated donor [MMUD], them had died. One of 10 patients had died because of grade 4 n ¼ 10), were enrolled in this study. All these patients met the aGVHD. Median follow-up post- HSCT for surviving patients was diagnostic criteria for HLH. 1680 days (91-3269 days). Relapse rate and non- relapse mortality Except one patient all others were treated initially with chemo- rate were 20% and 10% respectively. Median progression free based on the use of etoposide, dexametha- survival and median overall survival were 1668.5 days (68- 3269 sone and cyclosporine-A (CSA). Disease status at time of HSCT days) and 2634 days (391- 4641) respectively. was known as remission for 15 patients, only one patient had Conclusion: Our results suggest that allogeneic HSCT provide active disease. a curative treatment for PMF patients. Myeloablative regimens The year 2010 was a corner-stone for this study. After this date is seems to be effective and safe for especially younger PMF we use fludarabine in conditioning regimen and CSA-MTX patients. combination in GvHD prophylaxis. Disclosure of Interest: None declared. Results: Of the 16 HSCT recorded in 8 years period, 8 (%50) were performed after 2010.Consanguinty was reported in 8 patients (%50). Neurological involvement at diagnosis was found in 9 patients (%56). The median time interval from diagnosis to HSCT was 24,8 months (range, 7-96). A Haemoglobinopathy and inborn errors of conditioning regimen based on fludarabinewas adopted in 8 metabolism HSCT (%50). CSA alone (7 patients) or in combination with methyl-prednisolone (one patient), or in combination with methotrexate (8 patients) was used for GvHD prophylaxis. The median number of nucleated cells infused was 9x108/kg AB142 (range, 2.6-14.4) for bone marrow recipients, and 5.1x107/kg Hematopoietic stem cell transplantation for (range, 0.2-15) for cord blood recipients, while the median hemophagocytic lymphohistiocytosis: a single center number of CD34 cells was 13.5x106/kg (range 5.2-22.1) for experience peripheral blood stem cell recipients. In 13 patients neutrophil K. Yalc¸ın1,A.Ku¨pesiz1,*, F. Tayfun2, N. Eker1, V. Uygun3, engraftment was obtained at a median time of 18.2 days M. Akcan4, G. Karasu3, V. Hazar5, M. A. Yes¸ilipek3 (range 13-33) and platelet engraftment was obtained at a 1Pediatric Hematopoietic Stem Cell Transplantation Unit, Akdeniz median time of 36 days (range 14-86). In all of the patients, 5 University School of Medicine, Antalya, 2Pediatric Hematology- had veno-occlusive disease (VOD) and four of them underwent Oncology, Diyarbakır Children Hospital, Diyarbakır, 3Pediatric HSCT before 2010. Eleven of 16 (%69) children are alive and in Hematopoietic Stem Cell Transplantation Unit, Bahc¸es¸ehir complete remission. University School of Medicine,˙ Istanbul, 4Pediatric Hematology- Conclusion: We confirm that HSCT represents a curative Oncology, Adnan Menderes University School of Medicine, Aydın, treatment for HLH. The major problem about HSCT is still TRM. 5Pediatric Hematopoietic Stem Cell Transplantation Unit, Medipol We observed a trend, although not statistically significant, University School of Medicine,˙ Istanbul, Turkey towards an improvement in TRM after 2010. We think that this trend is associated with fludarabine based conditioning Introduction: Hemophagocytic lymphohistiocytosis (HLH) is a regimen. syndrome of pathologic immune activation. There are two Disclosure of Interest: None declared.

S615 AB143 Results: Median follow up was 17 months (range 1–101 Outcomes of allogeneic unrelated hematopoietic stem cell months). Overall survival - 71%. 12 patients are alive. The transplantation with reduced intensity conditioning in median time to neutrophil recovery 40,5x109/L was children with inherited metabolic disorders 19 days (9-25). All but 1 patients were engrafted with A. S. Borovkova1,*, N. Stancheva1, A. Ratz1, P. Kozhokar1, achievement of full donor chimerism on D þ 30. The S. Razumova1, E. Semenova1, O. Paina1, L. Zubarovskaya1, median alpha-L-iduranidase (AIDU) activity in leucocytes in B. Afanasyev1 HS patients was 61,3 nM/mg/18 h on D þ 30 and 77,6 nM/mg/ 1R.M.Gorbacheva Memorial Institute of Children Oncology, 18 h on D þ 100 (normal AIDU activity-61,0-175,5 nM/mg/18 h). Hematology and Transplantation, FIRST PAVLOV STATE MEDICAL In 4 cases we observed decrease of donor chimerism UNIVERSITY, St.Petersburg, Russian Federation from day þ 60 to day þ 180. In 3 patients donor lympho- cytes infusions were administered due to decreasing Introduction: Inherited metabolic diseases (IMD) lead to donor chimerism. In 1 case full donor chimerism was significant morbidity and death in childhood in the majority achieved. Three patients still have mixed chimerism (40-69%) of affected patients. Allogeneic haematopoietic stem cell without signs of disease progression. The incidence of graft transplantation (alloHSCT) is a possible treatment option for failure and transplant related mortality was 6.9% and 19% several metabolic disorders. Myeloablative regimens (MAC) are respectively. One patient with osteopetrosis developed associated with a lot of treatment related toxicity that limits its pulmonary venoocclusive disease. One patient had severe usage in pts with multisystem involvement. Late effects of mucositis (stage III according WHO classification). Grade III-IV MAC such as infertility and growth retardation impair long- aGvHD had 4 patients, chronic GVHD - 3. Patient with term quality of life. Reduced intensity conditioning regimens metachromatic leucodystrophy died on D þ 37 from (RIC) are more attractive in this group of patients because they progression of disease. One patient died from TRALI-syndrome are associated with lower rates of severe toxicity. The aim of on day þ 45. Two patients died from grade IV aGVHD this study was to evaluate efficacy of allo-HSCT with RIC in of gut. children with IMD. Conclusion: In our study we demonstrate that RIC in children Materials (or patients) and methods: From 2006 to 2014 16 with IMD is rather effective, well tolerated and is the promising children with IMD (Hurler syndrome (HS) - 8, Krabbe disease - alternative to MAC in this group of patients. Further study is 2, metachromatic leucodystrophy - 1, X-linked adrenoleuco- needed to evaluate long-term complications of RIC. dystrophy -1, malignant osteopetrosis – 2, adenosine-deami- Disclosure of Interest: None declared. nase-deficiency – 1, Farber disease type II - 1) undergone alloHSCT. Matched (n ¼ 10) or partially matched (8-9/10) (n ¼ 6) unrelated HSCT were performed. Peripheral blood AB144 stem cells (PBSC) (n ¼ 10) and bone marrow (n ¼ 6) were used Allogeneic MSD-Bone Marrow Tranplantation in Globoid as source of hematopoietic stem cells. All patients received Cell Leukodystrophy (Krabbe‘s disease) in a Small Infant conditioning regimen consisted of Fludarabine 150 mg/m2, C. Salvador1,*, R. Crazzolara1, B. Meister1, G. Kropshofer1 Melphalan 140 mg/m2 and ATG 60 mg/kg. CsA plus short 1Pediatrics I, Medical University Innsbruck, Innsbruck, Austria course of methotrexate or MMF were used for GVHD prophylaxis. In 3 cases we used CD34 þ -selection or CD3/ Introduction: Globoid cell leukodystrophy is a genetically CD19 þ -depletion of PBSC for prevention of GVHD. Detailed transferred lysosomal storage disorder of childhood. It is patient and transplantation characteristics are presented in rapidly progressive by a deficiency in galactocerebrosidase, table 1. which leads to accumulation of cytotoxic lipids damaging

[AB143]

S616 oligodendrocytes and Schwann cells and leading to demye- Mycophenolate Mefotil.He had a second HSCT from same lination of central and peripheral nervous system. One sibling donor bone marrow with a dose of 9.0x108/kg.Condi- possible therapy for this disease is umbilical cord blood tioning was with BU 16 mg/kg /CY total 200 mg/kg(but CY transplantation before showing symptoms, although this 100 mg/kg days þ 3 and þ 4).Had a long period of aplasia treatment is not curative and it only decreases progression with 22 units of irradiated red cells concentrates and 34 units of symptoms in some patients. of irradiated platelet concentrates. Materials (or patients) and methods: We here report a Results: He had rejection after the first and second HSCT but currently six months old girl, which was diagnosed with regenerated self at þ 42 and þ 63 days respectively with no globoid cell leukodystrophy at the age of three weeks. This GVHD. Developed iron over load that responded to oral iron early diagnose was possible due to an older sibling who died chelator. Pre -transplant stable haemoglobin was 7 g/dl but of this disorder at the age of 8 months. Our patient had an despite the rejection after 1 year post HSCT patient developed HLA-identical sibling donor available and we started con- a high stable persistence of fetal haemoglobin(29.5%),HBA2 ditioning regimen, including Fludarabine, Treosulfan and 2% and HbS 68.5%.Current stable Hematocrit is 30%,Hemo- Thiotepa, at the age of four weeks. On day -1 the patient globin 9.8 g/dl, Total White blood cell count 9200/ul and developed seizures, which were not able to interrupt with platelet 182,000/ul.Patient is clinically stable with very few common medication. She was transferred to our pediatric insignificant crises. intensive care unit, was intubated and finally seizures stopped Conclusion: A sickle cell anaemia patient who had two after 72 hours. MRI imaging showed marked edema oft the HSCT and rejected both grafts but selectively developed brain and lesions of basal ganglia. Bone marrow was high persistence fetal haemoglobin. He is clinically stable transfused on day 0, GvHD prophylaxis was accomplished with an improved haemoglobin concentration and other with Metothrexate, ATG and Cyclosporine. haematological parameters compared to the pre- transplant Results: The patient was extubated five days after start of state. seizures. Complications after transplantation were marked oral Disclosure of Interest: None declared. mucositis and skin toxicity due to conditioning regmien as well as GvHD grade I-II oft he skin. The neurological situation AB146 presented initially after extubation with ophisthotonus and Ethnopharmacological survey of herbal medicines used shrill crying, improved however after start of Baclofen for the treatment of sickle cell anaemia in Chhattisgarh, orally. The patient showed engraftment on day 23 with full India donor chimerism. She did not show any further seizures S. Patra1,*, B. Anju2, P. Patra3 under medication with Levetiracetam. MRI imaging shows 1 atrophy of central nervous system, but the child develops Department of Botany, Govt. N.P.G. college of Sciences Raipur, India, Raipur, 2Department of Botany, Bhilai Mahila Mahavidya- enjoyably. 3 Conclusion: We here describe a MSD-bone marrow laya, Bhilai, Department of Biochemistry, Pt. JNM medical transplantation in a small infant with globoid cell leukody- College, Raipur, India strophy, which was diagnosed at the age of three weeks due to a index case in the family. One day before bone marrow Introduction: Sickle cell Disease is an autosomal recessive transfusion the child developed a status epilepticus, leading to genetic disorder with a wide variety of complications. admission at intensive care unit. It is not yet clear, if these Sickle cell anemia has no widely available cure, hydroxy- seizures and the resulting neurological situation can be urea is the only FDA-approved medication that focused explained by the underlying disease or by the treatment on augmenting fetal haemoglobin production, providing toxicity or by both. the exogenous RBCs through repeated blood transfusions Taken together we show, that allogeneic MSD-bone marrow is for patients not responsive to hydroxyurea therapy, or a good alternative for transplantation in globoid cell replacing the deficient bone marrow through trans- leukodystrophia. plantation. All of the above treatments have unfavorable side References: Maria L. Escolar et al, N Engl. J Med, 2005 effects, such as myelosuppression of blood cells from Staba et al., N Engl. J Med, 2004 continuous use of hydroxyurea, iron overload from repeated Disclosure of Interest: None declared. blood transfusions, or immunosuppressive treatments required to sustain a bone marrow transplant. As there are several limitations on current drug therapies, alternative Ideal AB145 therapies with anti-sickling activity and similar degree of High persistence fetal Haemoglobin after primary efficacy without the troublesome side effects have been rejection of Allogeneic Hematopoietic Stem Cell researched extensively. Increased incidence of the adverse Transplantation for a Sickle cell disease patient: A case effects and economic burden in the usage of modern system report of medicine may be a reason for the interest in traditional N. G. Bazuaye1,* medicine. 1Department of Haematology, University of Benin Teaching Materials (or patients) and methods: Traditional healers Hospital, Benin, Nigeria have utilized other and similar herbs to combine into similar formulations to treat sickle, with clinical success. In order to Introduction: Sickle cell Disease (SCD)patients with high determine the extent of use of herbal medicines, we persistence fetal haemoglobin have been reported to have a conducted a cross-sectional survey in Rahud village of Balod better clinical outcome. Allogeneic HSCT is the only curative district from Chhattisgarh state. During a 6-month period, therapy but not without complications of rejection villagers, middle man and local people were directly asked to especially with reduced intensity conditioning(RIC) than full know about the plants used by the villagers, middle man and myeloablative therapy. We report the only patient in our local people. Further, information about the different herbal transplant centre who had rejection after 2 consecutive formulation used by the local people for treating sickle cell HSCT but developed a stable persistence of fetal anaemia was documented. Haemoglobin. Results: During this ethnobotanical survey, 44 herbal and food Materials (or patients) and methods: Master MA is 12 years plants used by tribal communities of Chhattisgarh for treating old student diagnosed in infancy with SCD. He was on sickle cell anaemia were identified. The vernacular name, hydroxyurea for 3 months only before the first HSCT. He useful parts, ecological distribution and medicinal uses of the had a first HSCT in 2012 from his matched identical sibling identified plants were reviewed. bone marrow with a stem cell dose of 5.7x108/kg body Conclusion: The use of herbal drugs has seen a rapid weight. Conditioning was with FLU160mg/m2 /BU 14 mg/kg increase as they provide wide biological activity, have good and immune suppression ATG(Horse),Cyclosporin A and safety margins and are available at affordable cost.

S617 Although some of the herbs and their formulations used in the Results: The 2 years overall survival (OS) was 70% treatment of sickle cell anemia are known for its antioxident and the progression free survival (PFS) was 60%. While, and anti-sickling activities, the exact bioactive compound(s) and remission status less than complete remission (CR) determine their structure(s) and their toxicity is to be delineated. emerged to be a poor prognostic factor for OS in univariate Disclosure of Interest: None declared. analysis, high b2m levels and comorbidity indices revealed to be independent poor risk factors for both OS and PFS. The present study demonstrated that even if the patient is in CR before ASCT if he has high b2m, the 2 years OS decreases from 100% to 49% (Figure 1). Moreover, Lymphoma lymphopenia for the first time was demonstrated to predict PFS in ASCT in NHL patients. Conclusion: Our findings suggest that b2m at transplantation predict the outcome after ASCT in NHL and further investiga- AB147 tion with larger sample sizes is warranted. Beta-2 Microglobulin Predicts The Outcome After Disclosure of Interest: None declared. Autologous Stem Cell Transplantation in Non-Hodgkin Lymphoma E. Birtas Atesoglu1,*, A. Hacihanefioglu1, Z. Gulbas2 AB148 1Department of Hematology, Kocaeli University, 2Bone Marrow Allogeneic Stem-Cell Transplantation for advanced Transplantation Center, Anadolu Medical Center Hospital, mycosis fungoides/Sezary syndrome: Single-center Kocaeli, Turkey experience E. Atilla1,*, D. Sahin2, P. Ataca1, S. Civriz Bozdag1, S. K. Toprak1, Introduction: Autologous stem cell transplantation (ASCT) is M. Kurt Yuksel1, P. Topcuoglu1, B. N. Akay3, H. Sanli3, G. Gurman1, an established therapeutic modality in the treatment of M. Ozcan1 lymphomas, especially in the relapse setting. In the 1ANKARA UNIVERSITY DEPARTMENT OF HEMATOLOGY, 2ANKARA present study, we aimed to define pretransplantation factors UNIVERSITY DEPARTMENT OF INTERNAL MEDICINE, 3ANKARA including Beta-2 microglobulin (b2m) that influence outcomes UNIVERSITY DEPARTMENT OF DERMATOLOGY, ANKARA, Turkey following ASCT in patients with Non-Hodgkin lymphoma (NHL). Introduction: Several studies have suggested that allogeneic Materials (or patients) and methods: We analyzed retro- hematopoietic stem cell transplantation (HSCT) may improve spectively 78 NHL patients who had undergone ASCT from survival in advanced MF and SS. We aim to present the August 2010 to January 2013.Detailed information concerning outcome of the patients with MF or SS who underwent HSCT. patients presentation at diagnosis as well as the disease status, Materials (or patients) and methods: We retrospectively comorbidity indeces and other variables of known prognostic evaluated a total of 8 patients, 5 with advanced stage MF and importance were evaluated before the initiation of condition- 3 with SS who underwent peripheral HSCT rescue between ing chemotherapy for AHSCT. Oct 2012 and Oct 2014. All patients have received at least

[AB147]

S618 [AB148]

Table 1. Patient characteristics, transplant characteristics and outcomes

Age/ Diseaseonsetto ClinicalStage Disease Large Number Conditioning Regimen (MA/ Acute Chronic Outcome Sex/ HSCT status cell of prior RIC)/ GVHD prophylaxis GVHD/ GVHD/ Disease (years) at HSCT Trans- treatments Grade location formation

53/M/ 11 T4N2M1B0 Active - 4 RIC (Flu þ Cy þ TBI þ ATG) þ /2 NA Ex,fungal infection,23th MF IVB disease CsA þ MMF dayposttransplant 50/M/ 2 T4N2M0B2 Active þ 8 RIC (Flu þ Cy þ TBI) þ /3 Limited Relapsedafter 10 months, MF IVA1 disease CsA þ MMF ECP 61/M/ 4 T3N1M0B0 Active þ 4 RIC (Flu þ Cy þ TBI þ ATG) þ /2 Extensive VGPR (8months) MF IIB disease CsA þ MMF 55/M/ 3 T4N2M0B1 Active - 3 RIC (Flu þ Cy þ TBI þ ATG) 0 - Relapsedafter 4months SS IIIB disease CsA þ MMF 56/F/ 3 T4N1M0B0 Active þ 5MA(Cyþ TBI) CsA þ MMF þ /2 - Relapsed 4 months MF IIIA disease 50/F/ 1 T4N2M0B2 Active - 6 RIC (Flu þ Cy þ TBI) NA - Ex, fungalinfection, 16th SS IVA1 disease CsA þ MMF þ PREDNOL dayposttransplant 49/M/ 4 T4N0M0B0 Active - 7 RIC (Flu þ Cy þ TBI) 0 - Ex, acinetobactersepsis MF IIIA disease CsA þ MTX 16th dayposttransplant 49/M/ 3 T4N2M0B1 Active - 3 MA (Cy þ TBI) CsA þ MMF 0 Limited CR (24 months) SS IIIB disease three lines of therapy prior to HSCT (Table 1). Two out of 8 relapsed/refractory Hodgkin and non-Hodgkin lymphoma patients were transplanted from unrelated HLA mismatched patients who required allogeneic HCT. donors and other 6 patients had fully matched siblings. All of Materials (or patients) and methods: We retrospectively the patients had refractory disease prior to HSCT, 6 of 8 analyzed 46 relapsed/refractory lymphoma patients (25 NHL; patients received reduced intensity conditioning regimen 21 HL) who underwent allo HCT rescue treatment Ankara (Fludarabine-Cyclophosphamide-TBI þ /-ATG) while 2 patients University Department of Hematology BMT Unit. Chi-square test received myeloablative conditioning regimens including Cy- was used in comparison between two groups due to engraft- TBI. Peripheral blood was the stem cell source in all the ment kinetics, post-transplant complications, post-transplant patients. For graft-versus host disease prophylaxis, patients responses. Po.05 was considered statistically significant. received cyclosporine plus methotrexate or mycophenolate Results: The median age of the patients in NHL group was mofetil except one patient in whom prednisolone was used 40,2±14,2 and 28,8±7,9 in HL group. Peripheral blood was due to renal toxicity. the preferred stem cell source in both NHL and HL patients Results: The median age was 59 years (range 49-61). The (24/25, 17/21). Sixteen of twenty five non Hodgkin lymphoma median time from diagnosis to HSCT was 3 years (range 1-11 patients received myeloablative conditioning regimen years). The Sorror HCT-CI scores prior to HSCT were listed as: in whereas reduced intensity conditioning was the prefered 2/8 score of 0, 3/8 score of 1, 3/8 score of 2. The patients conditioning regimen in HL group. Also, transplantation from achieved engraftment at a median of 12 days after HSCT (range unrelated donor was also significantly higher than the NHL 0-22). Two of the patients died of the early transplant-related group. Fifteen patients had primary refractory disease in both mortality due to fungal or Acinetobacter infection. Two groups. Median follow-up period from transplantation was 26 complete remission, 2 very good partial remission (VGPR), and months (range 4-98) in 46 patients.Complete remission was 2 minimal responses were seen in surviving 6 patients. A VGPR observed in 5/21(23%) patients in NHL group while 9/21(42%) converted into CR after GVHD development. Acute and cGvHD patients in HL group.Engraftment could not be achieved in 5 developed in 57.1% of the patients (4/7) and in 3 out of 5 NHL patients and 3 Hodgkin lymphoma patients due to death patients surviving longer than 100 days after the transplanta- in aplasia period.Median time for neutrophil engraftment was tion. The follow-up duration was between 16 days and 769 14 days and platelet engraftment was 15 days for the Hodgkin days. The possibility of two-year progression free and overall lymphoma wheras non-Hodgkin lymphoma neutrophil and survival was 37.5%±1.7% and 62.5%±1.7%, respectively. platelet engraftment median were both 16 days. Acute graft Conclusion: Our results in a small patient group suggest that versus host disease(GVHD) was detected in 13/25(52%) allo-HSCT is an effective treatment in patients with advanced patients in NHL group and 10/21(47%) patients in HL group. stage refractory MF/SS. One year overall survival in NHL group was detected as 40% in Disclosure of Interest: None declared. NHL group while 55% in HL group. Conclusion: Patients with HL received reduced intensity AB149 conditioning regimen more frequently and transplantation Allogeneic Stem-Cell Transplantation for Hodgkin and was mostly from unrelated donors. In our retrospective study, non-Hodgkin Lymphoma: Single-center experience the allogeneic stem cell transplantation outcomes were found 1,* 2 1 1 1 to be similar with the previous reports. E. Atilla , T. Aydin , M. Merter , P. Ataca , S. Civriz Bozdag , Disclosure of Interest: None declared. S. K. Toprak1, M. Kurt Yuksel1, P. Topcuoglu1, O. Arslan1, M. Ozcan1, T. Demirer1, O. Ilhan1, H. Akan1, M. Beksac1, N. Konuk1, G. Gurman1 AB150 1ANKARA UNIVERSITY DEPARTMENT OF HEMATOLOGY, 2ANKARA Previous autograft does not increase the early TRM after UNIVERSITY DEPARTMENT OF INTERNAL MEDICINE, ANKARA, allogeneic HSCT for patients with relapsed/refractory Turkey lymphoma E. Klyuchnikov1,*, A. van Randenbourgh1, H. Lellek1, D. Janson1, Introduction: Although long term survival rates can be S. Ro¨mer1, S. Heidenreich1, C. Wolschke1, N. Kro¨ger1 achieved with chemoradiotherapy approaches in lymphoma, 1Department for stem cell transplantation, University Hospital, one third of the patients can relapse during the follow up. Hamburg, Germany Allogeneic hematopoietic stem cell transplantation (HCT) remains to be the only curative treatment approach for Introduction: The role of allo-HSCT in patients with relapsed/ relapsed/refractory lymphoma patients.Here, we present our refractory lymphoma still remains controversial. It provides a

S619 tumor-free graft and possible graft-versus-lymphoma effect as number of platelet transfusions was significantly lower in well as increased toxicity in comparison to autologous HSCT. lipegfilgrastim group (11 vs 19 units; P ¼ 0.03). We also Whether the use of allo-HSCT might be associated with observed higher infection rate in the lipegfilgrastim group increased transplant-related mortality and might have a (80% vs 45%; Po0.01), but with a milder grade (1/2 according negative impact on survival is unclear. to CTCAE v.4) compared to filgrastim. The median length of Materials (or patients) and methods: Here, we analyzed hospitalization was significantly shorter in patients receiving outcomes for 43 patients (male, n ¼ 36, 84%) with median age lipegfilgrastim compared to filgrastim (26 vs 40 days; of 57 years (range 22-72) who underwent an allo-HSCT with P ¼ 0.000). (n ¼ 15) or without (n ¼ 28) previous autografting. The ‘‘non- Conclusion: Lipegfilgrastim is safe and effective as filgrastim auto’’ patients experienced a trend to have higher Karnofsky in hematopoietic reconstitution and significantly reduces the index (75% vs.67%), more chemorefractory disease (57% vs. number of platelet transfusions and the length of hospitaliza- 33%) and to receive more reduced-intensity allografts (57% vs. tion in patients with lymphoproliferative malignancies after 40%). The median time between auto- and allo-HSCTs was 11 auto-SCT. months (range 3-202). Disclosure of Interest: None declared. Results: After median follow-up of 17 months (range 2-44), a total of 17 patients were alive. The 2-year probability of OS and DFS were 43% (95% CI: 20-66%) and 32% (95% CI: 7- AB152 57%), respectively. The TRM at 100 days was 8% in ‘‘non-auto’’ Long-term clearance of blood and bone marrow and 7% in ‘‘auto’’ group. The cumulative incidences of TRM involvement after graft failure and autologous and relapse at 1 year after allo-HSCT were: ‘‘non-auto’’: reconstitution following reduced-intensity allogeneic stem 13% and 57%; ‘‘auto-group’’: 13% and 46%, respectively cell transplantation in a patient with Se´zary syndrome: a (P40.05). case report Conclusion: The previous auto-HSCT was not associated with G. Saporiti1,*, D. Vincenti1, E. Berti2, E. Tagliaferri1, C. Annaloro1, increased early TRM after allo-HSCT and should not be a B. Motta1, A. Cortelezzi3, F. Onida3 reason to avoid an allo-HSCT also in elderly patients. A trend to 1Hematology, 2Dermatology Unit, Fondazione IRCCS Ca’ Granda decreased relapse incidence in the "auto" group remains to be Ospedale Maggiore Policlinico, 3Hematology, Fondazione IRCCS further investigated. Ca’ Granda Ospedale Maggiore Policlinico - University of Milan, Disclosure of Interest: None declared. Milan, Italy

Introduction: Sezary syndrome (SS), a rare cutaneous T cell AB151 lymphoma, is characterized by the presence of erythroderma Efficacy and safety of lipegfilgrastim in patients with with pruritus, lymphadenopathy and atypical circulating lymphoproliferative malignancies after autologous stem lymphocytes. Its clinical course is generally aggressive, with cell transplantation only 25% of patients surviving beyond 5 years from diagnosis. E. Stefanko1,*, J. Rybka1, I. Deren´-Wagemann1, M. Biedron´1, Allogeneic hematopoietic stem cell transplantation (allo-HSCT) D. Urbaniak-Kujda1, J. Dybka1, R. Poreba˛ 2, K. Kuliczkowski1, is currently the only treatment option with curative intent in T. Wro´bel1 patients with SS. 1Department of Hematology, Blood Neoplasms and Bone Materials (or patients) and methods: On June 2008 a 53 Marrow Transplantation, 2Department of Internal, Occupational years-old gentleman was diagnosed with MF and treated with Diseases and Hypertension, Medical University, Wroc"aw, Poland phototherapy until April 2011, when lymphocytosis in peripheral blood occurred and a Sezary clone was detected Introduction: Recombinant granulocyte-colony stimulating by flow cytometry (CD4 þ ,CD7 þ ,CD26-) both in peripheral factors (G-CSFs) are commonly used to accelerate bone blood and in bone marrow. A total body CT scan was negative marrow recovery and reduce the duration and incidence of for lymphadenophaties and visceral involvement. Therefore chemotherapy-induced neutropenia after autologous stem ECP was performed from April 2011 to April 2013 achieving cell transplantation (auto-SCT). Lipegfilgrastim is a novel glyco- only partial response. From September 2012 to January 2013, pegylated G-CSF effective in reducing neutropenia in patients while a search for a HLA-matched unrelated donor was started with breast cancer receiving myelosuppressive cytostatics. for a possible allogeneic allo-HSCT treatment strategy, low There is no data concerning efficacy of lipegfilgrastim in dose cyclophosphamide per os was also introduced with no patients after autoSCT. The objective of the study was to clear benefit on the disease control. Therefore, on June 14th evaluate the safety and impact of lipegfilgrastim administra- 2013, following a non-myeloablative conditioning regimen tion on haematopoietic reconstitution and supportive care in including pentostatin 4 mg/msq (day -10 and -3), TBI 200 cGy patients with lymphoproliferative malignancies after auto-SCT. (day -1) and ATG 2,5 mg/kg for 3 days (day-4,-3,-2), the patient Materials (or patients) and methods: We analyzed prospec- underwent allo-HSCT from a 9/10 HLA unrelated male donor. tively 30 patients (12 women and 18 men; median age 50±13 GvHD prophylaxis included cyclosporine and mycophenolate years) including 13 with multiple myeloma (MM), 5 with mofetil. Hodgkin’s lymphoma (HL) and 12 with non-Hodgkin’s Results: During the early post transplant phase the patient lymohoma (NHL). The median number of transplanted showed progressive neutropenia complicated by an Acineto- CD34 þ cells was 3,96±1,56 106/kg body weight. Patients bacter iwoffi bacteremia on day þ 14. Chimerism analysis received a single 6 mg dose of lipegfilgrastim on day þ 1 after performed on CD8 þ peripheral lymphocytes in the same day transplantation. The results were compared with historical unveiled 100% of recipient DNA. On the following days, control (14 women and 19 men; median age 49±6 years) instead of showing the expected hematopoietic recovery, including 13 patients with MM, 8 with HL and 12 with NHL. leukocyte count continued to decrease; a bone marrow This group was treated with filgrastim (n ¼ 33) at a dose 5 mg/ examination performed on day þ 24 failed to show the kg/d on day þ 1 posttransplant and continued to an absolute presence of any donor engraftment at chimerism analysis, neutrophil count (ANC) 41,5x109/L. while showing a 23% of lymphocyte infiltrate with aberrant Results: The median time to ANC40,5 109/L reconstitution phenotype at flow cytometry (CD4 þ ,CD7 þ ,CD26-). On day in lipegfilgrastim and filgrastim groups was 13 and 11 days þ 28 the patient was diagnosed with graft failure and respectively (P ¼ 0.460). There was also no differences in the immunosuppressive therapy was discontinued. Autologous median time to platelet engraftment420 109/L between neutrophils (4500/mmc) and platelet recovery (450000/ two study groups (15 vs 16 days; P ¼ 0.70). The median mmc) were observed on day þ 45 on day þ 32, respectively. consumption of filgrastim vials a 0,3 mg was 19. No differences The diffuse skin lesions remained stable during the whole in the duration of febrile neutropenia were seen between transplant period. Following discharge, the patient underwent lipegfilgrastim and filgrastim-treated patients. The median regular outpatient visits and blood/marrow examination,

S620 showing normal CBC counts with persisting lymphocytopenia The engraftment of CB is said to be lower than that of bone (o500/mmc) in blood and no Sezary cells by flow cytometry in marrow transplantation. However our study shows the bone marrow. On May 2014, due to cutaneous disease engraftment of CB is not bad and transplantation of single progression, TSEB was started (36 Gy) achieving a very good CB is feasible in patients with even heavily treated. partial remission. At the last follow up visit on November 14th Disclosure of Interest: None declared. 2014, the patient was still in blood and marrow CR with minimal skin disease. Conclusion: Because allogeneic graft failure in patients with AB154 haematological malignancies is normally associated with High dose chemotherapy with autologous stem cell disease recurrence or progression, especially in the setting of transplantation for relapsed Hodgkin lymphoma: Single reduced intensity conditioning, this case appears to be tertiary care centre experience thought-provoking with regard to its correct interpretation, J. Zekri1,2,*, M. Kelta2, E. Abdelghany2,3, J. Ur Rehman2, Z. Khan4, suggesting possible immunological mechanisms as the M. Aslam2, M. Bayoumy2, R. Al-Saadi5, R. Jaleesah2, D. Reyad2,6 responsible for both graft rejection and the long-term 1College of Medicine, Al-Faisal University, 2Oncology, King Faisal clearance of blood and marrow disease involvement. Specialist Hospital (Jeddah), Jeddah, Saudi Arabia, 3Oncology, Disclosure of Interest: None declared. National cancer institute, Cairo University, Cairo, Egypt, 4Radiology, 5Research centre, King Faisal Specialist Hospital (Jeddah), 6Al-Faisal University, Jeddah, Saudi Arabia AB153 The engraftment of cord blood transplantation for Introduction: One large randomized trial showed that lymphoma in high risk high dose chemotherapy (HDC) and autologous stem cell H. Kodo1,* 1 transplantation (ASCT) provided extended Event Free Hematology, Metropolitan Tama Medical Center, Tokyo, Japan Survival (EFS) for patients with relapsed Hodgkin’s lymphoma (HL). As other prospective randomized studies have proved Introduction: The engraftment is the first obstacle of cord difficult to accrue, reports from retrospective series can add blood transplantation. valuable information. This study reports our single centre We transplant the cord blood transplantation in patients with experience. lymphoma and studied the influences of conditioning regi- Materials (or patients) and methods: All patientsZ15 years mens, HLA, the characteristics of cord blood on the old with relapsed and refractory HL who underwent HDC & engraftment. ASCT from June 2001 until December 2013 were included. Materials (or patients) and methods: 35 patients with non- Data was collected retrospectively by the investigators. Hodgikin’s lymphoma (high risk 31, standard risk 4) were Relevant imaging was reviewed and reported by an indepen- transplanted with single cord blood. 8 patients were dent radiologist. transplanted because of relapse after allogeneic cord blood Results: 54 patients were identified and reviewed. Median age transplantation(1) or autologous peripheral blood stem cells was 22 (15-49) years. 26 were males and 28 were females. 48 (7). Regarding conditioning regimen 23 of reduced intensity of (89%) patients underwent HDC & ASCT after achieving conditioning (RIC) and 12 of myeloablative conditioning radiological response to salvage chemotherapy. The rate of (MAC). GVHD prophylaxis were mainly Cy-A/MMF for RIC complete radiological response to salvage chemotherapy was regimen or Cy-A/MTX for MAC regiemn. Patients with first and 13% and improved to 44% within 3 months after ASCT. second CR were divided into standard risk and the rest were all After median follow up of 25 months, 31 (57%) are still alive into high risk. with no evidence of relapse or progression. Median EFS Results: The mean of nucleated cell count (NCC) 2.15 x 10^7, was 24 months (95% CI: 8.7-39.3), 3 years EFS was 56% CD34-positive cells 0.74 x 10^5 were transplanted. Regarding and median overall survival was not reached. Stage at first the compatibility of HLA 9/35 cases were less than 2 loci, 26/35 relapse, extranodal disease, bulky mediastinal disease at were not. All patients were statistical analyzed by Kaplan Meier relapse did not significantly impact EFS on uni-variant and Method in JMP. multi-variant analyses. There was a trend for longer EFS (30 vs. Results: A patient with NK/T lymphoma after autologous 23 months; P ¼ 0.18) in patients with longer time from end of PBSCT and relapse was died of multi-organ failure on day 2. first line treatment until relapse (412 vs. r12 months) The rest of all patients were survived more than 28 days after respectively. One hundred day transplant related mortality transplantation and 28 patients engrafted by day 28 and 6 was was 5.5%. not. The rate of engraftment was 81%. Conclusion: HDC & ASCT for relapsed HL is safe. Our findings Regarding the characteristics of cord blood there was no are consistent with published phase III results. Longer follow difference according to NCC. CD34-positive cells and CD34/ up is warranted. NCC ratio also made statistically difference on engraftment. Disclosure of Interest: None declared. They shows quick and good engraftment. HLA disparity more than 2 loci did not any influences on the engraftment of cord blood. Other factors such as conditioning regimen (MAC vs AB155 RIC), GVHD prophylaxis did not make difference. Disease status Rituximab, thiotepa, busulfan, and cyclophosphamide did not influence on engraftment. No other factors including (R-TBC) conditioning for pre-transplantation PET-positive CB characteristics and patient status make a significant primary refractory or relapsed Hodgkin lymphoma difference. L. Gopcsa1,*, P. Remenyi1, A. Barta1, A. Batai1, Z. Csukly1, Conclusion: Only one was observed as early death and the J. Fabian1, L. Lengyel1, G. Mikala1, M. Reti1, E. Torbagyi1, rest of all were survived more than 28 days in spite of most T. Masszi1 patients in high risk. That is there is no factors which influence 1St Istvan and St Laszlo Hospital, Budapest, Hungary on early death This is probably due to our conditioning regimens is enough to prevent rejection and not so strong Introduction: In relapsed Hodgkin lymphoma (HL), negative that damage patients. PET before the use of BEAM conditioning for autologous Many reports say usually NCC is one of the important factors haematopoietic stem cell transplantation (AHSCT) is associated to get engraftment. Our study do not. Probably NCC include with favourable long-term outcome. BEAM remains the mononuclear cells which is lymphocytes, monocytes and standard high dose chemotherapy (HDC) combination for HL nucleated red blood cell and cd34-positive cells. Because the despite its suboptimal results in patients with refractory most important cell fraction is CD34-positive cells, it is very tumours or poor prognostic features, with long-term reasonable that the ratio of CD34-positive cell/NCC and CD34 event-free survival rates of 50% to 15%. PET scanning appears cells are good index of engraftment. to be an accurate tool for assessing prognosis in patients

S621 who are eligible for AHSCT. Given poor outcomes with median interval from diagnosis to transplantation was 39 BEAM, intensification of treatment with 3-alkylating agent months (range, 8 to 147 months). At the time of trans- combination such as R-TBC (rituximab, thiotepa, busulfan, plantation, 6 patients were in complete remission, 3 in cyclophosphamide) has been developed. The TBC partial remission and 6 in progression/relaps of lymphoma. conditioning is efficaciously used for central nervous system All patiens underwent autologous transplantation, lymphomas. We adapted this regimen for eligible HL patients 3 (20%)patients had received brentuximab vedotin as a bridge with pre-transplantation PET active tumours. In addition, to allogeneic transplant. The conditioning regimen for incorporating brentuximab vedotin (BV) in the treatment transplantation consisted of fludarabine 120 mg/m2 plus strategy may improve the therapeutic efficacy of HDC and melphalan 140 mg/m2 and cyklosporin A plus methotrexate AHSCT. as GVHD prophylaxis. Materials (or patients) and methods: In this retro- Results: Overall survival (OS) and disease free survival (DFS) spective study, we reviewed the outcomes of all consecutive were 58,3% and 36,6% at 1 year. One-year transplant-releated patients treated with AHSCT using R-TBC conditioning mortality was 32% (15% at day 100). Acute GVHD developed in with or without postgraft BV for primary refractory and 46% (III-IV 23%), chronic GVHD developed in 38%. relapsed HL. Conclusion: The role of allogeneic SCT in HL is still a subject of Results: Seven patients (5 men and 2 women) were treated controversy. Allogeneic stem-cell transplantation can improve with the median age of 23 years (range:20-36). All cases the outcome for patients with Hodgkin’s disease if transplan- were HL nodular sclerosis including 6 patients with primary tation is performed early after relapse. refractory disease and 1 patient with partially chemo- Disclosure of Interest: None declared. sensitive relapse. Salvage therapy before AHSCT was complex, with median number of 2 regimen including 2 AB157 patients who received BV. Unfortunately, all patients had Autologous Stem Cell Transplantation in Elderly Patients PET-positive active tumours at AHSCT. The median time from (4 ¼ 60 years) with Myeloma or Lymphoma: Single Center diagnosis to AHSCT was 21 months (range:11-48). R-TBC Experience regimen started on day -10, with rituximab (500 mg/m2). On 1,* 1 1 1 1 days -9 to -7, patients received thiotepa (250 mg/m2/die), on M. Gunduz , E. Atilla , P. Ataca , G. Pekcan , S. Civriz Bozdag , S. K. Toprak1, M. Kurt Yuksel1, P. Topcuoglu1, O. Arslan1, days -6 to -4, busulfan (0,8 mg/kg i.v. every 6 hours/die), and 1 1 1 1 1 on days -3 and -2, cyclophosphamide (60 mg/kg/die) was M. Ozcan , T. Demirer , O. Ilhan , H. Akan , M. Beksac , N. Konuk1, G. Gurman1 given. The median day to neutrophil engraftment and platelet 1 recovery were 9 (range:8-10) and 10 (range:9-16), respectively. ANKARA UNIVERSITY DEPARTMENT OF HEMATOLOGY, ANKARA, Toxicities were significant. All patients had neutropenic fever, Turkey severe mucositis, GI-toxicities. There was 3 cases of engraft- ment syndrome. One patient had a documented CMV- Introduction: Significant therapeutic advances have recently pneumonitis. Four of 7 patients developed toxicoderma. The been made in the care of older patients in myeloma and 100- day transplant-related mortality rate was 0%. Following lymphoma. Although there are many elderly patients who are AHSCT, three patients received 4 cycles of BV for maintenance. eligible for high dose therapy (HDT) with autologous stem cell At this early follow-up (median 12 months, range 3-14 transplantation (ASC) rescue, the evidence is inadequate. months), all patients are alive in PET negative complete Therefore we aimed to analyze the efficacy and safety of HDT metabolic remission (CMR). with ASC rescue in our elderly myeloma and lymphoma Conclusion: In this current analysis, we report our early patients. institutional experience of HDC/AHSCT with R-TBC condition- Materials (or patients) and methods: Between Jan 2003 and ing for primary refractory and relapsed HL. Our patient Oct 2014, 133 patients (84 M; 49 F) admitted to Ankara University population had been heavily pretreated. R-TBC is a safe and Department of Hematology were included in the study. Their efficacious conditioning regimen even in pretransplant PET diagnosis were as following; 116 plasma cell dyscrasia (114 positive cases. Interestingly, all patients achieved CMR after multiple myeloma, 1 plasmositoma and 1 primary amyloidosis) R-TBC/AHSCT. More follow-up is needed to determine the and 17 non-Hodgkin Lymphoma (10 Diffuse large B cell, 6 long-term outcome. It is possible, that R-TBC with BV Mantle cell and 1 follicular). We retrospectively evaluated the maintenance may further improve outcome compared to early toxicity and outcome of the HDT. standard BEAM conditioning in patients with refractory Results: Median age of the group was 63 years (60-74 ys) disease with pretransplant PET-positivity. In addition, larger while median time from diagnosis to the transplantation was cohort of patients is needed to evaluate the role of R-TBC 11.4 months (3.1-188.7 months). Most of the patients had low conditioning following AHSCT with post-autograft BV in Sorror Comorbidity Index (CI) (68.4%). In comparison of the refractory or relapsed HL. toxicities including oral mucositis, diarrhea, renal failure, liver Disclosure of Interest: None declared. and cardiac toxicity and the incidences of overall infections, we found only oral mucositis and diarrhea that were more frequent in patients with myeloma than lymphoma (P ¼ 0.01 AB156 and 0.03, respectively). When similar comparisons were Allogeneic stem-cell transplantation for Hodgkin repeated according to age (o65 ys vs Z65 ys), Sorror CI lymphoma single-center experience (low vs intermediate and high), the intensity of conditioning L. Mohamamdova1,*, P. Jindra1, K. Steinerova1 regimen for melphalane (standard vs low dose), the rates of 1University hospital Pilsen, Hematooncology, Pilsen, Czech toxicities and infection did not change. In addition, engraft- Republic ment kinetics were not affected by the diagnosis and age. However, faster neutrophil recovery was observed in low and Introduction: Hodgkin lymphoma (HL) is one of the most intermediate Sorror CI compared to high index (median 11 curable lymphoma. Current treatment modalities can cure up days vs 12 days, P ¼ 0.03). Early-transplant related mortality to 80% of patients. However, 20-30% will experience relapse, (TRM) was seen only in 5 patients. The diagnosis, age and and 5-10% will not respond to primary chemotherapy at all. Sorror CI were not found to have any impacts on both TRM The prognosis for relapsed or refractory HL is poor. Allogeneic and overall survival. hematopoietic stem-cell transplantation (HSCT) may improve Conclusion: Although the oral and gastrointestinal mucositis the outcome. were more frequent in myeloma patients, TRM and OS did not Materials (or patients) and methods: This study examines differ. Therefore, HDT with ASC rescue was demonstrated to the outcome of 15 patients who underwent nonablative be a safe option in the treatment of myeloma and lymphoma allogeneic HSCT for HL over a 10-year period at a single patient in advanced age in our cohort. center. 15 patients (median age 37 years, range 23 to 56). The Disclosure of Interest: None declared.

S622 AB158 non-Hodgkin lymphoma(NHL), and 10% to 15% of pediatric Hematopoietic stem cell transplantation for T-cell NHL. In the report of ’Retrospective study of children with lymphoma: the experience of the Tunisian national center malignant lymphoma in Korea’, which analyzed total 517 of hematopoietic stem cell transplantation children who were diagnosed as lymphoma from 1991 to R. EL FATMI 1,*, N. BEN ABDELJELIL1, L. TORJEMANE1, I. HAMDI1, 2000, 11.2% of the patients were diagnosed as Hodgkin A. LAKHAL1, S. LADEB1, T. BEN OTHMAN1 lymphoma while others were NHL. ALCL comprises about 10% 1Tunisian national center of hematopoietic stem cell of NHL in children in the report, this fraction is known to be transplantation, Tunis, Tunisia similar in the western countries. The optimal treatment of ALCL is controversial whether the intensive short pulsed Introduction: Peripheral T cell lymphoma (PTCL) is a therapy or the prolonged intermittent therapy is better. Until heterogeneous group of non -Hodgkin lymphoma with now, both ways have similar prognosis of 60B75% in disease generally poor outcome with conventional chemotherapy. free survival. Materials (or patients) and methods: We retrospectively Materials (or patients) and methods: We retrospectively analyzed data from 26 patients transplanted for peripheral T reviewed the children younger than 16 years old who were cell lymphoma (PTCL), from January 2008 to September 2014. newly diagnosed and treated as ALCL at five institutions in Results: Seventeen patients underwent autologous transplanta- Korea from 1994 to 2013. We collected the data of clinical tion (AUSCT) and 9 others underwent allogeneic transplantation characteristics, modality of the treatments, and treatment from matched sibling donor (ALSCT).The median age was 45 outcomes. years (range; 20-62), 17 patients were male( 65%).The median Results: Total number of the patients is seventy six, time from diagnosis to transplantation was 13,7 months (range fifty(65.8%) were male while female patients account 34.2%. 3,9 -51,7). The most frequent histological subtype was PTCL-NOS Their median age at diagnosis was 10.6 years(range, 0.3-16.0 (n ¼ 11, 42%), followed by anaplastic large cell lymphoma ALK- years). Advanced stage ALCLs were more common. stage III (n ¼ 6), NK/T-cell lymphoma (n ¼ 3), hepatosplenic T cell and IV were sixty patients, 78.9%. ALK mutation was studied lymphoma (n ¼ 3), angioimmunoblastic T-cell lymphoma only in sixty five patients. Among studied patients, fifty (n ¼ 2) and enteropathy-associated T-cell lymphoma (n ¼ 1). three(81.5%) resulted in positive. B symptoms were shown in At diagnosis, 19 patients (73%) were classified as high 50% of the patiehts. Lymph node was the most common initial intermediate or high risk according to AA-IPI. involvement site(84.2%). Mediastinum, bone, visceral, lung, At transplantation, 12 patients (46%) were in complete and bone marrow involvement were common. Five(6.6%) remission (CR1 ¼ 7 and CR2 ¼ 5), 11 (42%) patients were in patients received high dose chemotherapy(HDC) followed by partial remission (PR1 ¼ 7 and PR2 ¼ 4) and 3 (12%) patients autologous hematooietic stem cell transplantation(HSCT) as have progressive disease. primary treatment. Their age at diagnosis were below 10 In the AUSCT group, all the patients received BEAM (carmus- years(median age, 6.8 years), and all of the five patients were tine, etoposide, cytarabine and melphalan) as a conditioning advanced stages. There was no relapse among them, while regimen. In the ALSCT group, patients received total body only one patient died due to treatment related mortality. irradiation (TBI) associated to etoposide in the 3 cases and to Median follow up duration of the patients is 67.3 months. The fludarabine in 3 cases. Busilvex-cyclophosphamide was used in 5-year overall survival(OS) rate of the patients who underwent 1 patient and was associated to fludarabine in 2 patients. HSCT as primary treatment was slightly lower than the other All patients achieved engraftment. In the AUSCT group, among seventy one patients(80.0±17.9% vs 88.2±4.0%, P ¼ 0.6). the 6 patients transplanted in CR, 4 maintained their remission However 5-year relapse free survival(RFS) rate was and 2 relapsed. Of the 8 patients transplanted in PR, 3 achieved higher(100.0% vs 80.9±5.3%, P ¼ 0.39). Thirteen patients CR, 1 maintained PR and 4 progressed. The disease continued to experienced relapse(17.1%), the median time of relapse from progress in the 3 patients transplanted in progression. In the initial diagnosis was 48.8 months. Nine of the thirteen relapsed ALSCT group, among the 6 patients transplanted in CR, 5 patients(69.2%) underwent HDC followed by HSCT(auto-HSCT maintained their remission and 1 relapsed. Of the 3 patients 7, allo-HSCT 2). Patients suffering from relapsed ALCL still transplanted in PR, 2 achieved CR and 1 progressed. showed 76.2±12.1% of survival rate. Among relapsed After a median follow up of 7, 7 months (range 1,5-78,3 patients, the 5-year OS rate of the nine patients who transplant months) for all patients and 18,5 months (range 3,9-76,6 after relapse was significantly better than four patients who months) for surviving patients, the 3-year overall survival (OS) are without transplant(100.0% vs 25.0±21.7%, P ¼ 0.003). and progression free survival (PFS), were (50%/46%), (41%/ Conclusion: The optimal treatment strategy for pediatric 35%) and (66%/66%), respectively in all the patients, the ALCL in Korea is not yet defined, but overall chemotherapy AUSCT group and the ALSCT group. There were no cases of response for pediatric ALCL shows favorable prognosis. HSCT TRM in the AUSCT group and 1 patient (11%) died from GVHD in high risk ALCL patients is feasible primary treatment to in the ALSCT group. decrease relapse rate, and can improve survival rate for Conclusion: Despite the poor prognosis of PTCLs, AUSCT and relapsed ALCL. However, acute toxicities and late effects ALSCT offer a favorable survival outcome and an acceptable related to transplantation remain to be the problem. Along toxicity, especially for patients in complete remission. with the HDC followed by autologous/allogeneic HSCT, Disclosure of Interest: None declared. immunotherapy with anti-CD30 antibodies, Brentuximab, is expected to significantly improve the prognosis of ALCL patients in the future. AB159 Disclosure of Interest: None declared. Experience of High Dose Chemotherapy and Hematopoietic Stem Cell Transplantation of Anaplastic Large Cell Lymphoma in Korea AB160 S.-M. Hahn1,*, H.-S. Kim1, J.-W. Han1, K.-N. Koh2, H.-J. IM2, Safety of biosimilar filgrastim in patients with lymphoma J.-J. Seo2, H.-J. Kang3, K.-D. Park3, H.-Y. Shin3, J.-W. Lee4, undergoing neutropenia-inducing chemotherapy: a N.-G. Chung4, B. Cho4, C.-J. Lyu1 sub-analysis of the NEXT study 1Pediatric Hematology and Oncology, Severance Hospital, Yonsei S. Lepreˆtre1,*, F. Maloisel2, D. Kamioner3, C. Berthou4, H. Albrand5 University Health System, 2Pediatric Hematology and Oncology, 1Henri Becquerel Hospital, Rouen, 2Sainte-Anne clinic, Strasbourg, Asan Medical Center, 3Pediatric Hematology and Oncology, 3Hoˆpital Prive´ de l’Ouest Parisien, Trappes, 4Hoˆpital Morvan, Seoul National University Hospital, 4Pediatric Hematology and Brest, 5Hospira France, Meudon La Foret, France Oncology, Catholic Medical Center, Seoul, Korea, Republic Of Introduction: The safety and efficacy of biosimilar filgrastim Introduction: Anaplastic large cell lymphoma(ALCL) is a (Nivestimt, Hospira Ltd), a granulocyte-colony stimulating rare tumor, which accounts for approximately 3% of adult factor licensed for the treatment of chemotherapy (CT)-

S623 induced febrile neutropenia (FN), were assessed in the NEXT dose dependent manner especially in the T cell tumors. We study. Here we present the results for patients receiving CT for used the ATG as the part of the conditioning regimen in the all lymphoma. patients and to evaluate the long-term anti-leukemia effect, Materials (or patients) and methods: The NEXT study was a the safety and complication in the patients with relapsed or prospective, observational, non-interventional, longitudinal, refractory PTCL national, multicentre study conducted in France which Materials (or patients) and methods: 7 patients( male 3, recruited 2114 adult patients undergoing cytotoxic CT and female 4) were enrolled into this study. Median patient receiving biosimilar filgrastim as primary or secondary age at the time of transplantation was 30 years (range, prophylaxis (ppx), or curatively. The primary objective was to 27–53 years). At the time of transplant, no patient evaluate the safety of biosimilar filgrastim. Secondary objec- reached first or subsequent complete response (CR) with tives were to evaluate the efficacy of biosimilar filgrastim, to conventional therapy or the salvage therapy. The median describe characteristics of patients treated with, and patterns courses number of prior therapies was eight (rang, four to of use of, biosimilar filgrastim. Data recorded included pt fifteen). characteristics, biosimilar filgrastim treatment-related data, Donors were 10/10 HLA matched related (4), 10/10 and treatment emergent AEs. Pts were monitored for 1–6 CT matched unrelated (2) and 8/10 matched unrelated (1). cycles at three visits: inclusion, during treatment, and The median number of CD34 þ cells within the allografts following CT. was 8.9/kg body weight (BW) (range,5.85–24.85/kg BW). Results: Of 2114 pts enrolled in the NEXT study, 2102 pts The conditioning regimen in this study consisted of were included in the overall analysis. Of these, 406 pts Rabbit antithymocyte globulin (ATG 2.5 mg/kg 4 days), were diagnosed with lymphoma (Hodgkin’s [n ¼ 55], non- total-body irradiation (10 Gy in five fractions), cyclo- Hodgkins [n ¼ 351]). On inclusion, the mean age±standard phosphamide (60 mg/kg 2 days) and teniposide (15 mg/ deviation (SD) of lymphoma pts was 63.3±16.3 years (62% kg 2 days) in all patients. Graft-versus-host disease (GVHD) male). 90.7% of pts had a performance status of 0 or 1, and prophylaxis was cyclosporine based, usually in combination 47.3% were at Ann Arbour stage 4. The most common CT with methotrexate. Quantitative chimerism analyzes were protocol was ‘R-CHOP’. The majority of pts (98.8%) received performed using short-tandem-repeat-based polymerase biosimilar filgrastim as ppx (88.6% primary ppx, 11.4% chain reaction techniques at regular intervals for every 4 secondary ppx). weeks after transplantation in bone marrow at the first six Of the group receiving biosimilar filgrastim with curative months. intent: 60% had an infection; mean treatment duration±SD Results: All patients achieved a sustained complete remission was 5.2±1.5 days; 40.0% received a dose of 30 MIU; and after allogeneic HSCT. They remained alive without evidence biosimilar filgrastim was administered subcutaneously in all. In of disease. The median duration of PFS reached to 41(7-95) those receiving biosimilar filgrastim as ppx, the median time to months. The longest survivor was a female patient who initiation of biosimilar filgrastim was 6 days after last dose of relapsed post autologous HSCT. Three patients experiened CT; mean treatment duration±SD was 6.6±4.0 days; 72.5% with acute GvHD grades II and one patient gradually envolved received a dose of 30 MIU; and biosimilar filgrastim was into cGVHD. The high levels of CMV DNA were detected in six administered subcutaneously in all. patients in the first three months after transplant. One of them At least 1 AE was experienced by 21.5% of pts. The most gradually envolved into viral haemorrhagic cystitis. One common AEs (45.0% of pts) were bone/muscular disorders patient suffered from bacterial pneumonia. But there was also (16.0%) and muscle pain (15.5%). Anti-infective ppx was no patient dying from cytomegalovirus infection or the other reported in 42.9% of pts. In the group receiving biosimilar infectious complications. There was no case of venous filgrastim as ppx, 7.1% (95% confidence interval [CI]: 4.9, 10.1) occlusive disease. experienced FN (primary ppx: 6.4% [95% CI: 4.3, 9.63]; Conclusion: ATG may be improve the outcome of secondary ppx: 12.2% [95% CI 4.86, 26.02]). Infection occurred allogeneic hematopoietic stem cell transplantation in 4.3% (95% CI: 2.66, 6.8) of those on ppx. in patients with relapsed or refractory peripheral T-cell In this analysis, 7.1% of pts were hospitalised for FN and/or lymphoma. infection. The mean duration of hospitalisation±SD for FN References: 1.Rudiger T, Weisenburger DD, Anderson JR and/or infection after the first CT cycle was 11.4±17.4 days. et al. Peripheral T-cell lymphoma (excluding anaplastic 2.8% of pts had a CT dose reduction, and 5.9% had a delay in large-cell lymphoma): results from the Non-Hodgkin’s administration of CT due to FN and/or infection. Lymphoma Classification Project. Ann Oncol 2002; Conclusion: Biosimilar filgrastim was effective and well- 13:140–149. tolerated in pts undergoing CT for lymphoma and is an 2. Grullich G, Ziegler C, Finke J et al. Rabbit Anti T-Lymphocyte alternative therapeutic option for pts with CT-induced Globulin Induces Apoptosis in Peripheral Blood Mononuclear neutropenia. Cell Compartments and Leukemia Cells, While Hematopoietic Disclosure of Interest: S. Lepreˆtre: None declared, F. Maloisel Stem Cells Are Apoptosis Resistant. Biol Blood Marrow Funding from: Hospira, Sandoz, Pfizer, D. Kamioner: None Transplant, 2009, 15:173-182. declared, C. Berthou: None declared, H. Albrand Employee of: 3. Huixia Liu, chun wang, Youwen Qin,et al. Polyclonal Rabbit Hospira SAS. Antithymocyte Globulin induces apoptosis and has Cytotoxic Effects on human Leukemic Cells. Clinical Lymphoma, Myeloma AB161 & Leukemia 2012,12: 345-54 Anti-thymocyte globulin(ATG) could improve the outcome Disclosure of Interest: None declared. of allogeneic hematopoietic stem cell transplantation in patients with relapsed or refractory peripheral T-cell AB162 lymphoma High-dose etoposide(VP16) in mobilization for patients Y. Jun1,*, W. chun2 with refractory lymphoma 1hematology, Shanghai Jiaotong University affiliated Shanghai Y. Cai1,*, C. Wang2, J. Jiang2 First People’s Hospital, shangha, 2hematology, shanghai, China 1hematology, 2hematolgy, Shanghai General Hospital, Shanghai, China Introduction: Overall survival (OS) at 5 years in PTCLs is not higher than 25–30% with the exception of ALK þ ALCL cases Introduction: The patients with aggressive lymphoma who Patients with relapsed or refractory peripheral T-cell lymphoms have a poor prognosis and unlikely to be cured with are generally considered incurable with conventional thera- conventional chemotherapy. We used high-dose etoposide pies. The early experiment result in our hospital showed that in mobilization followed auto-SCT for refractory lymphoma ATG inhibited the proliferation of lymphoid tumor cells in a and shared our experience.

S624 Multiple myeloma

AB163 Outcome In Multiple Myeloma Patient After Autologous Stem Cell Transplantation, response to transplant B. Acosta-Maldonado1,*, M. Alfaro1, V. Valencia-Carlo1, L. M. Valero-Saldan˜a1, S. Rivas-Vera1 1Hematology, Instituto Nacional de Cancerologia, Mexico city, Mexico

Introduction: High-dose therapy followed by autologous stem cell transplantation (ASCT) has been the standard frontline consolidative therapy for patients with newly diagnosed multiple myeloma (MM). The complete remission (CR) rate increased from 25.9% (7/27) before ASCT to 70.4% (19/27) following ASCT (Po0.01). The probability of OS for 5 years was 52.2% for the patients in the ASCT group and 33.1% for those in the non-ASCT group (P40.05). We analyzed the results of 63 patients with multiple myeloma who underwent autologous stem cell transplantation (ASCT). Materials (or patients) and methods: We evaluated 63 consecutive individuals with Multiple Myeloma, referred and treated at Instituto Nacional de Cancerologı´a, between January 2003 to December 2013. High-dose melphalan (200 mg/m2) was used for conditioning regimen. The characteristic for response to transplant were analyzed by Pearson’s test. Kaplan–Meier method was used to estimate OS probabilities, with differences compared by the log- test. Results: It was analyzed the results of 63 patients (30 males and 33 females) with multiple myeloma who underwent ASCT. The median age of patients was 50 years (range, 31–66 years). 26 Materials (or patients) and methods: 40 patients (median patients (41.3%) had at least one comorbidity. Diagnosis was age 33 (13-61) years) with refractory Non-Hodgkin’s lymphoma performed in our hospital in 55.6% of cases, the rest was (NHL, n ¼ 32) or Hodgkin’s lymphoma (HD, n ¼ 8) received referred in complete response to ASCT from other health high-dose etoposide (VP16 10-15 mg kg-1 d-1 2d) in institutions. The majority of patients were in Durie Salmon Stage mobilization in our center. Remission status prior to mobiliza- IA and IIA (31.7% and 39.7%, respectively). Patients with Stage I tion was PD (n ¼ 40). The use of such granulocyte colony- (International Staging System) were 57.1%. Before transplant, stimulating factor (G-CSF, 5-10 mg/ kg-1 d-1) mobilized per- patients received treatment using thalidomide þ dexametasone ipheral blood stem cells (PBSC) after high-dose etoposide until (n ¼ 47), VAD (vincristine, adriamycin and dexamethasone, the end of leukapheresis. Peripheral blood Stem cell was n ¼ 6), other (n ¼ 10). 33 patients were treated with combined collected and frozen in 80 1 refrigerator. All these patients treatment modality (chemotherapy plus radiotherapy). Overall received auto peripheral blood stem cell transplantation (auto- response after ASCT was 84.1%; complete response 36 (57.1%), PBSCT). Conditioning regimen was BEAM (n ¼ 19, 47.5%) or very good partial response:11 (17.5%), and partial response: 6 CBV (n ¼ 21, 52.5%). (9.5%). Thirty patients (47.6%) were underwent to double ASCT Results: Twenty-eight patients (70%) were assessable for (Tandem) while only 33 patients a single transplant is response after high-dose etoposide at a median follow-up of performed. Tandem transplant not show improving response 39days (range 17-172 days), 12 patients (30%) had no or survival. Bivariate analysis showed that ISS, less than 12 response. Median follow-up of 28(4-66)months, Sixteen month from diagnostic to the ASCT were statistical significant patients(40%) reached CR after auto-PBSCT. 15 for response to treatment (Po0.05). Durie-Salmon, combined of the 28 patients (53.6%) who had response to high-dose treatment modality, fever and neutropenia and low seric etoposide reached CR, 4 patients (14.3%) reached PR, 9 albumin were important for overall survival (Po0.05). Ten years patients (32.1%) succumb to progression of disease. 1 of the overall survival from diagnosis was 59%. Overall survival from 12 patients (8.3%) who had no response to high-dose ASCT to death was 67% to 9 years. etoposide reached CR, 1 patients (8.3%) reached PR, 10 Conclusion: The transplantation goal is to reinforce the patients (83.4%) succumb to progression of disease. response achieved by induction therapy and improve PFS The estimated 1-year OS and EFS were 69% and 56.7% and OS. In this review, an early ASCT showed long survival. respectively, 2-years OS and EFS were 63% and 52% One limitation of this study is the lack of adequate information respectively. The prognosis of the patients who in relation initial staging, valuation of prior response to HCT had no response to etoposide was poor. The estimated and follow up of referred patients. The emergence of novel 1-year OS and EFS were 25% and 1[nn1] 6.7% respectively. agent-based therapy combined with ASCT has revolutionized Two group of comparison differences have statistics MM therapy by improving the CR rates and OS, raising significance (Po0.01). questions concerning the role of hematopoietic stem cell Conclusion: High-dose etoposide could be used in transplantation in this setting. However, In our population the refractory lymphoma as rescue therapy in mobilization. use novel agents it still limited because economic issue. So It can increase the EFS and OS of patients who had currently ASCT remains as the standard of care in newly response. The hematopoietic stem cells collection diagnosed multiple myeloma. and hematopoietic reconstitution are not affected by References: L Kumar etal. High-dose chemotherapy with etoposide. autologous stem cell transplantation for multiple myeloma:- Disclosure of Interest: None declared. what predicts the outcome? Experience from a developing country. Bone Marrow Transplantation (2009) 43, 481–489. Disclosure of Interest: None declared.

S625 AB164 Introduction: Autologous stem cell transplantation (ASCT) are Abstract Withdrawn standard for young patients (pts) with multiple myeloma (MM). The aim of the study was to investigate the blood coagulation status in MM pts during ASCT. AB165 Materials (or patients) and methods: 10 pts: 6 males, 4 females Early versus delayed autologous stem cell transplantation at the age of 27–65 years (median 56) were included in the study. for multiple myeloma in patients receiving induction After induction therapy and successful peripheral blood stem cell therapy with bortezomib mobilization 6 of them achieved CR, 3 – VGPR and 1 – PR. 2 I. Osipov1, E. Darskaya1,*, E. Babenko1, V. Vavilov1, B. Afanasyev1 Melphalan (Mel) in dose 100 mg/m was used as a conditioning 1Raisa Gorbacheva Memorial Institute of Children Oncology, regimen during 2 days. ASCT was conductedonthe2-ddayafter Hematology and Transplantation. First Pavlov State Medical Mel infusion. A central venous catheter (CVC) was inserted on the University of St. Petersburg, St.-Petersburg, Russian Federation day before Mel administration and heparin-sulphate continuous infusion (500 IU/hour) as thromboprophylaxis was started till Introduction: Despite the use of new drug (proteasome engraftment. Hemostasis analysis was performed 5 times: before inhibitors, immunomodulatory drugs), autologous hemato- (point 0) and 24 hours after heparin administration (point 1), on the next day after Mel infusion (point 2), once during the period poietic stem cell transplantation (ASCT) is the standard 9 treatment for young patients with multiple myeloma (MM). when WBC count was less, then 1 10 /l (point 3) and by the Materials (or patients) and methods: We retrospectively time of engraftment (point 4). Condition of hemostasis was reviewed the outcome of patients with newly diagnosed MM. assessed by the results of activated partial thromboplasin time 60 patients were included in the analysis from January 2008 (APTT, normal rate: 25–38 sec) and thrombin generation test until October 2014. All patients received induction treatment (TGT), namely by using endogenous thrombin potential (ETP, with bortezomib (VD, CVD, PAD regimen chemotherapy) and normal rate: 760–1450 nM * min) in every point. first ASCT within 12 months from the start of treatment and for Hypercoagulation was considered in cases when APTTo25 sec 2 months after collection PSCT (early ASCT, n ¼ 35) or at a later and ETP41450 nM * min. Hypocoagulation was estimated by date (delayed ASCT, n ¼ 25). Median time to ASCT was 7.3 data APTT438 sec and ETPo760 nM * min. months vs. 15.4 months in early vs. delayed ASCT. The median Time series data were analyzed using the SAS TIMESERIES age the patients was 51,1 vs. 49,6 years. 71% of patients in the procedure. Statistical analysis was performed using SAS 9.3 (SAS early ASCT (n ¼ 25) and 80% (n ¼ 20) in the group with Institute Inc. Cary, NC). Significance level of alpha was set at 0.05. delayed ASCT received maintenance therapy with bortezomib Results: Before heparin administration (point 0) APTT and TGT or IMiDs after ASCT. Median follow-up was 36.8 months. evaluation showed normal coagulation in 8 pts (mean APTT Results: The 5-year overall survival (OS) from diagnosis was was 34 sec (95% CI 32–37 sec), mean ETP - 1041 nM * min (95% 73,0% vs. 87,6% in early and delayed ASCT, which was not CI 1005–1488 nM * min)). Hypocoagulation was indicated in 2 statistically significantly (P ¼ 0.18). case by APTT, at that time hypercoagulation was detected in 2 Progression-free survival (PFS) was not significantly different in pts by ETP. On the next day after starting thromboprophylaxis the groups of early and delayed ASCT. However, there is a (point 1), APTT and ETP have significantly (Po0,05) changed to tendency to increase the duration of PFS in patients, who hypocoagulability in response to heparin infusion: mean data achieved CR or VGPR after induction therapy and received APTT became 38 sec (95% CI 36–41 sec) and mean ETP - early ASCT. 3-yers PFS was 58% vs. 20% in the group early 894 nM * min (95% CI 701–1034 nM * min). After Mel (n ¼ 16) and delayed ASCT (n ¼ 20), P ¼ 0.11. The median time administration (point 2), APTT was statistically significantly to progression was, respectively, 47 m. vs 26.5 m. These decreased (Po0,05) reaching 34 sec (95% CI 33–36 sec). ETP differences are detected after 2 years of observation. Duration was also changed, the mean was 1109 nM * min (95% CI 929– of PFS did not differ in the group of early and delayed ASCT at 1229 nM * min). Hemostasis at point 3 was characterized by achievement a partial response. APTT mean 38 sec (95% CI 33–45 sec) and ETP mean 1002 nM * Disease-free survival (DFS) was better in the group with early min (95% CI 712–1103 nM * min). By the time of engraftment ASCT (n ¼ 13) than delayed ASCT (n ¼ 6). 3-year DFS was 68% we observed all values to be shifted towards the normocoa- vs 0%, respectively (P ¼ 0.06). gulation: mean values of APTT - 33 sec (95% CI 30–41 sec) and For early ASCT has been shown to increase PFS when using ETP - 1146 nM * min (95% CI 916–1176 nM * min). maintenance therapy (median PFS not achieved). In the group A CVC - related thrombosis occurred in 1 case. There was a without maintenance therapy the median PFS was 27.3 months. women of 56 y.o., with additional risk factors such as obesity (P ¼ 0.04). In delayed transplantation has not been shown the (BMI – 42) and heterozygous MTHFR polymorphism. The dose impact of maintenance therapy for the duration of PFS. of heparin was increased and folic acid therapy was started. Conclusion: Early or late ASCT is a good option for patients After 2 weeks recanalization was detected. with MM receiving induction treatment with bortezomib – Conclusion: We identify latent hypercoagulation status in 20% based therapy. pts of confirmed by elevated ETP. Those changes were Currently we have no evidence of the superiority of early ASCT, registered despite of achievement CR or PR MM. Thrombotic which may be related to the small number of patients and short complications have developed in 10% from our group. follow-up. Therefore, it is necessary to further observation. Notably, one should consider individual risk factors for each pt. The role of maintenance therapy after ASCT also requires Disclosure of Interest: None declared. clarification. Disclosure of Interest: None declared. AB167 Clinical outcomes of transplant-eligible multiple myeloma AB166 patients treated with frontline thalidomide/ Hypercoagulability in multiple myeloma patients dexamethasone undergoing autologous stem cell transplantation H. Ha1,*, J. Youk1, H. Park1, J.-O. Lee2, S.-M. Bang2, K.-H. Kim3,4, E. S. Urnova1,*, L. Mendeleeva1, O. Pokrovskaiya1, M. Gracheva2, J.-H. Park3,4, Y. Koh1,4, I. kim1,4, S.-S. Yoon1,4, S. Park1,4 I. Tarandovskii2, M. Nareyko1, E. Gemdjan3, M. Firsova1, 1internal medicine, Seoul national university hospital, Seoul, M. Soloviev1, L. Kuzmina4, E. Makunina1, E. Parovichnikova4, 2internal medicine, Seoul national nuiverstiy bundang hospital, V. Savchenko5 Seongnam, 3internal medicine, Seoul national university bor- amae medical center, 4Cancer Research Institute, Seoul national 1high dose chemotherapy of paraproteinemic hemoblastosis, university hospital, Seoul, Korea, Republic Of 2Biophysics laboratory, 3Biostatistics laboratory, 4bone marrow transplantation, 5Hematology Research Center, Russian Minis- Introduction: For successful treatment of transplant-eligible try of Health, Moscow, Russian Federation multiple myeloma (MM) patients, effective induction

S626 chemotherapy is crucial for long term disease control. auto-HSCT can successfully be performed in a developing Thalidomide in combination with dexamethasone (TD) is the country with results comparable to western literature. most prevalent regimen used as induction chemotherapy in Disclosure of Interest: None declared. Korea. The purpose of this study is to evaluate clinical outcomes of TD induction chemotherapy and to compare AB169 this outcome with other regimens including novel agents such Mobilization in multiple myeloma patients with plerixafor as proteasome inhibitor. affects graft cellular composition Materials (or patients) and methods: We retrospectively R. Fedele1,*, M. Cuzzola1, T. Moscato1, B. M. Oliva 1, A. Pontari1, reviewed 200 MM patient who received TD induction 1 1 1 1 1 chemotherapy between 2000 and 2014 at Seoul National A. Dattola , M. C. Cannata’ , A. Meliado’ , S. Barilla’ , C. Gareffa , E. Spiniello1, G. Console1, G. Messina1, G. Irrera1, M. Martino1 University Hospital and Seoul National University Bundang 1 Hospital. Baseline characteristics, tolerance of TD treatment, Azienda Ospedaliera BMM, Reggio Calabria, Hematology and and outcome of TD treatment were analyzed. Stem Cell Transplant Unit, Reggio Calabria, Italy Results: All patients (Male: Female 95:105) were below 65 years old at the time of diagnosis, and their median age was Introduction: Plerixafor (PLX) is a new mobilization agent that 57.1 years old (range 33.1-64.9). By International Staging is approved for use in combination with G-CSF in poor System, disease stage was I in 30.5%, II and III in 36.0% and mobilizer patients affected by multiple myeloma (MM) and 30.5%, respectively. Patients received median 4 cycles (range, non-Hodgkin lymphoma (NHL) and scheduled for high-dose 1-26) of TD as induction. Overall response rate was 78% (154/ chemotherapy and autologous hematopoietic stem cell 200) with proportion of patients achieving complete remission transplant (HSCT). There are lack of data on the graft content and very good partial response was 4.5% and 24.0% other than CD34 þ cell dose in patients mobilized with a respectively. Eighteen of patients experienced disease pro- combination of chemotherapy, G-CSF and PLX. gression despite of TD induction. Overall, 36.0% of patients Materials (or patients) and methods: We studied 3 patients (72/200) received salvage reinduction chemotherapy (Borte- affected by IgG lambda MM in partial remission after 4 cycles zomib containing regimen in 51 patients). With salvage of VTD, mobilized with conventional cyclophosphamide (CTX) treatment, 56.9% (41/72) patients showed objective response. 4 g/mq þ G-CSF 10 mcg/kg. After performing 2 apheresis Autologous stem cell transplantation was successfully per- collections insufficient to ensure a double procedure of HSCT, formed in 145 (72.5%) patients. Among these patients, we added subcutaneous PTX 0.24 mcg/kg reaching the proportion of patients who underwent autologous stem cell intended target of 5x10E6/kg CD34 þ cells respectively with transplantation after TD induction chemotherapy without further 2 collections in 2 patients and 3 collections in a patient. salvage induction chemotherapy was 69.6% (N ¼ 101). In each graft we evaluated total lymphocytes and their subsets Conclusion: TD as induction chemotherapy is quite efficient and monocytes by flow cytometric analyses. but not satisfactory compared to recently used regimen Results: Total lymphocyte content was gradually reduced including proteasome inhibitor in newly diagnosed MM from the first to the last apheresis regardless of PLX patients, and should be reconsidered for its role as frontline administration; the amount of T suppressor cells (CD3 þ / treatment. CD8 þ ), NK cells (CD3-/ CD16/56 þ ), B cells (CD19 þ ) was Disclosure of Interest: None declared. significantly higher in the grafts obtained after G-CSF þ PLX than after G-CSF alone. On the contrary, CD3 þ /CD4 þ cells were more representative in grafts obtained without PLX. The AB168 content of monocytes in different grafts resulted very HIGH DOSE CHEMOTHERAPY FOLLOWED BY interesting: it was reduced in the subsequent injections of AUTOLOGOUS STEM CELL TRANSPLANT FOR MULTIPLE G-CSF and is progressively increased with each consecutive MYELOMA dose of PLX (figure 1). N. Ali1,*, M. U. Shaikh1, S. Adil1 Conclusion: PLX determines modification of apheresis pro- 1The Aga Khan University, karachi, Pakistan duct respect G-CSF in terms of cellular types above all in terms of lymphocyte subsets and monocytes. Futher studies are Introduction: Autologous hematopoietic cell transplantation needed in larger group patients to confirm these data and to (auto-HCT) procedure is the standard of care for patients with analyze their impact on immune reconstitution, long-term multiple myeloma (MM) under the age of 65 years. Limited engraftment, patient outcomes after HSCT information is available from developing countries regarding Disclosure of Interest: None declared. the long term outcome of this disease after auto-HSCT. The objective of this study was to determine the overall outcome and complications in patient with multiple myeloma under- going auto-HSCT Materials (or patients) and methods: We performed auto- logous transplants using noncryopreserved and unmanipu- lated peripheral blood stem cells mobilized from the bone marrow to the peripheral blood using Filgrastim. High dose Melphalan (100 mg/m2/day) was used as the conditioning regimen. Results: From 2004-2014 a total of n ¼ 61 auto-HSCT were performed out of which n ¼ 10 were for MM. Two procedures were aborted due to insufficient stem cell dose. All patients had evidence of chemosensitive disease prior to transplant. There were n ¼ 7 males and n ¼ 3 females. The median age±SD was 48 years±7.5 (range: 32–55 years). All patients achieved engraftment. The median time to achieve 40.5 109/L granulocytes was 16 days. The overall stay in the unit ranged from 11-25 days. Grade III-IV mucositis was the most common complication. There was no transplant related mortality. After a median follow-up of 66.3 months (range: 43.8 -88.7 months) the overall survival was 70%. Conclusion: The overall survival was 70%. There was no transplant related mortality. Our results demonstrate that

S627 Solid tumours treatment. The patient underwent right hepatectomy and the histology was refined as a fetal HB. Post-surgery chemotherapy consisted of: Cisplatin 80 mg/m2 IV (24 h) inf þ alternating every 2 weeks with Carboplatin 500 mg/m2 þ Doxorobicine AB170 30 mg/m2 (2days) for 8courses. We use of high dose High dose chemotherapy and stem cell transplantation in chemotherapy with peripheral blood stem cell rescue as a childhood hepatoblastoma presenting with lung consolidation treatment. Autologous hematopoietic stem cells metastases were harvested in 1 day after Cyclophosphamide þ G-CSF A. Hedayati Asl1,* on behalf of Mehrvar A, Goodarzi K, stimulation. Apheresis successfully was done in patient Zangooei R, Emaam jomeh M weighing less than 10 kg. The conditioning regimen was: 1Pediatric Stem cell transplantation, MAHAK, Tehran, Iran, Islamic Etoposide 200 mg/m2 4 days, Caboplatin 400 mg/m2 4 days Republic Of and Melphalan 75 mg/m2 for 2 days. Results: Median time to absolute neutrophil count Introduction: Hepatoblastoma is a rare malignant neoplasm (ANC)40.5 109/L was 10 days, and the median time to of the liver, with an incidence of 0.5–1.5 per million platelet count420 109 was 13days. Toxicity was modest children, but it is the third commonest malignant abdo- and represented by mucositis. The patient was discharged 0n minal tumor in childhood following neuroblastoma and day þ 17. Today the patient remains disease-free, with normal nephroblastoma. The incidence of HB is most common in AFP levels and no radiological evidence of disease, 13 months infants and young children, with a median age at diagnosis following HDCT with peripheral stem cell rescue. of 16 months. The outcome of recurrent or metastatic Conclusion: Children with metastatic disease have fared hepatoblastoma is not always promising. Lung metastases poorly with 20%450% event free survival and new identified on diagnosis are one of the most reliable prognostic approaches for these patients remain desperately needed. indicators of an unfavorable outcome for patients with Patients with metastatic disease may benefit to use high dose hepatoblastoma. chemotherapy and stem cell transplantation. Longer follow-up Materials (or patients) and methods: We describe a one- is required to evaluate fully efficacy and long term survival of year- old male child affected by metastatic hepatoblastoma our patient. who achieved complete lung response only after conventional Disclosure of Interest: None declared.

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