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EBMT 2013

Stem cell biology R1470 Mobilization of PBSCs in heterozygous-for-β thalassemia donor by addition of plerixafor after failure of mobilization R1469 with G-CSF alone for (TcR) αβ T lymphocytes depletion in Traffi cking of CD34+ cells into the peripheral circulation haploidentical transplant in thalassemia patients during collection of stem cells by apheresis: experience of P Sodani (1), A. Lanti (2), M.L. De Simone (2), E. Fiorelli (2), a single centre K Paciaroni (1), G De angelis (1), C Alfi eri (1), A. Roveda (1), A. Isgrò (1), A. Dattola, R. Fedele, E. Massara, G. Cornelio, L. Imbalzano, G. Ressa, C Gallucci (1), M. Ribersani (1), L. Cardarelli (1), M Marziali (1), C. Garreff a, A. Meliadò, M. Cannatà, I. Callea, A. Pontari, E. Spiniello, F. Torelli (1), G. Adorno (2), O.m. Chiru (2), J Gaziev (1), G Lucarelli (1) P. Scaramozzino, D. Princi, G. Irrera (1)Mediterranean Institute of Hematology (Rome, IT); (2)Policlinico AO Bianchi Melacrino Morelli (Reggio Calabria, IT) Tor Vergata (Rome, IT)

Introduction: defi nition of standardized product is a product with Hematopoietic stem cell transplantation off ers the only chance of components having safe capacity of stable hemopoiesis reconsti- cure for patients with thalassemia. Haploidentical transplantation tution (effi cacy) without unseen eff ects or reactions (safety). Main may extend this possibility to the 50% to 60% of the patients who objective of all processing facilities is to have CD34+ cell products lack a suitably matched familial donor or an HLA-identical unre- standardized. To date, we did not have a consensus to defi ne the lated donor. Family members were assessed for HLA compatibility cell product, but we tried to standardize the mobilization’s capa- by serological methods or by high-resolution molecular analysis. city in aphaeretic collection products (A-HPC). Two heterozygous for β thalassemia family donor for the very Material and Methods: we evaluate n. 580 A-HPC and related sam- young son ( 4 years old and 7 years old respectively ) candidate ples. Quantifi cation/enumeration of CD34+ in absolute count was to haplo-SCT for thalassemia major did not achieve the expected performed with ISHAGE Protocol by fl ow cytometry FACSCalibur, target >8 x10 6 CD34+/kg recipient with conventional mobiliza- Stem kit and Trucount test tubes. lion (G-CSF 10 mcg/kg daily) and were successfully rnobilized with Results: CD34+ cells number in cell products collected by aphae- plerixafor. Following informed consent for off -Iabel use, the both resis exceeds 7,6 times CD34+ cells number calculated on periph- donors received a single subcutaneous injection of 24 mgs plerix- eral venous blood’s samples before collection, with statistical afor in combination with G-CSF with no side eff ects; 12 hrs later: signifi cance 0,00 (p=0,00). in the fi rst donor we observed circulating CD34+: 105/mcL (WBC Release’s calculation of CD34+ in pre-collection samples is 96 cell/ul 76x103/mcL), CD34/Kg recipient (post TcRα β T lymphocytes selec- (range 20-1404), in A-HPC is 975 cell/ul (range 64-17101). tion 11.2x106 CD34+/Kg,). In the second donor, using the same Collection’s performance for CD34+ (cell/ul) is 84% and it is inde- mobilization strategy the circulating CD34+ was 110/mcL (WBC pendent from processing of blood, diagnosis and conditioning 59.000x103/mcL), CD34/Kg recipient (post TcRα β T lymphocytes regimen. It depends from cellularity (cell/ul) on peripheral venous selection 8,1x106 CD34+/Kg,). The median time to neutrophil blood. and platelets engraftment was at 12 days and 14days, respec- Conclusions: Collected cells are present on peripheral venous tively.Using this new strategies in both haploidentical transplant blood at the begining of A-HPC collection. All patients show in thalassemia patients we observed more rapid immunological having a signifi cant increase of CD34+. A-HPC quality is directly reconstitution after 60 days post transplant, with an increase of dependent from peripheral venous blood quality. T cells (CD4+ and CD8+), B cells (CD19+) and NK cells (especially CD3-CD16+, with cytotoxic potential) in the peripheral blood of these patients. In conclusions: For all “poor mobilizers donors, included “ heterozygous for β thalassemia donor adequate transplant CD34+ cells dose was obtained by combination of Plerixafor with G-CSF with no side eff ects.

R1471 Autologous stem cell transplantation: analysis of haematimetric parameters R. Guerrero Camacho, M. Gasior Kabat, K. Humala Barbier, P. Gomez Prieto, R. De Paz, A. Marcos, R. Arrieta Gallastegui La Paz University Hospital (Madrid, ES)

Objectives: To investigate erythrocyte parameters oriented to program and optimize the apheresis procedure based on the stem cell harvest effi ciency. Materials and Methods: Medical records of 294 patients diagnosed with Non-Hodgkin Lymphoma (155), Hodgkin Disease (78) and Multiple Myeloma (61) were reviewed retrospectively. The apheresis procedures were conducted between 2000 and 2010. Effi ciency was calculated according to this formula: Effi ciency (%)=Obtained Mononuclear Cells (MNC)(109)/[( Total Processed Volume/Volemia) x (pre-apheresis MNC(109)+post-apheresis MNC (109)/2)].

S519 Studied parameters were: Haemoglobin (HB), Hematocrit (HT), concentration of 10% DMSO. For pediatric and clinically indicated Mean Corpuscular Volume(MCV), Mean Corpuscular Hemoglobin patients fi nal concentration is 5% DMSO. To assess the PBSCs’ (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC) quality, we evaluate total nucleated cells (TNC), CD34+ content and Red Blood Cell Distribution Width (RDW). These parameters and microbiological contamination (aerobic, anaerobic and myco- were compared between a Group with Effi ciency<60% and a logical cultures); an aliquot of each bag is stored (cell viability and Control Group with Effi ciency>80% using the Non-parametric clonogenic assays). Products are frozen in a rate-controlled pro- Mann-Whitney U-Test. grammed freezer (Nicool Plus PC, Air Liquide) and stored between Results: Patients with Hodgkin Disease showed a lower Effi ciency -150 and -196ºC in vapor and/or liquid phase of liquid nitrogen. than patients with Multiple Myeloma (p=0.015). In general, the At infusion time, frozen PBSC products are thawed at 37ºC and can Mean Effi ciency value of the patients studied was between 60 be directly infused (bedside thaw) or require post-thawed proce- and 80%, only a 6,5% of the patients (n=19) showed a Mean Effi - dures. Product’s quality is indirectly evaluated by engraftment’s ciency<60% and 48% showed a Mean Effi ciency>80%. Mean MCV success (Absolute Neutrophil Count – ANC). and MCH was lower in patients with Effi ciency>60% (p=0.026). Results: In 2011, 95 autologous PBSC from 69 patients (26% Conclusion: Pre-apheresis alterations in MCV, MCH and RDW pediatric – 5 months/16 years) were processed at our laboratory. values should be considered as adverse factors that can infl uence 58 PBSC were processed on the same day (61%) and the remain- the stem cell collection procedure. ing 37 (39%) were stored overnight at 4ºC and processed in the next morning. Average product volume was 128mL; 21 products (22%) required volume reduction. PBSC’s quality results: TNC average 3.34x108/Kg, CD34+ average 4.65x106/Kg and no microbiological contamination. 65/95 PBSCs collected (68%) were infused in 50/69 patients (72.5%); only 1/65 PBSC (1.5%) required post- thawing procedure (DMSO removal). 49 /50 patients (98%) recovered ANC between d+6 and d+17 (average 11.51 days). Conclusions: Results confi rm the quality of the new PBCS cryopreservation protocol.

R1473 Clinical and haematological features of oncohaematological patients with low content of CD34+ cells in autotransplant S. Gritsaev, A. Kuzajeva, I. Zapreeva, Z. Chubukina, S. Tiranova, V. Balashova, A. Seltcer, K. Abdulkadirov Russian Institute of Hematology (St.Petersburg, RU)

There are some conditions to get succesful engraftment after autologous stem cell transplantation (AutoSCT)and the main one is the number of CD34+ cells in the autotransplant which has to be ≥2x106/kg. The aim of the study was to characterize the patients with the autotransplant failure. There were 14 patients whose autotransplant contain less than 2x106 CD34+ cells/kg. For the control group 36 patients have been chosen. Leukoapheresis has been done with COBE Cpectra (version 6.1, Gambro). Median age of the patients was 47.5 years (19-62). There were next diagnoses: 6 patients with myeloma, 6 with nonHodgkin lymphoma and 2 with acute lymphoblastic leukemia. Before autologous stem cell transplantation (AutoSCT) the patients were treated with 1-4 regimens of chemotherapy. The mobilisation of stem cells was performed with 3 regimens. So the patients were divided into the 3 groups. In the fi rst group 5 patients were treated with G-CSF only. In the second group 8 patients were treated with Cph 1-3 g/m2 plus G-CSF. In the third group 1 patient was treated with high dose chemotherapy plus G-CSF. The median number of mononuclear cells R1472 in the autotransplants was 1.9x108/kg (1.0-5.8). The median Peripheral blood stem cell cryopreservation: a safety and number of CD34+ cells in the autotransplants was 0.51x106/kg quality analysis (0.03-0.72). C. Espadinha, A. Barbosa, M. J. Gutierrez, E. Pereira, M. Abecasis There was no any diff erence between study and control groups in IPO Lisbon (Lisbon, PT) the age, number of previous chemotherapy, total dose of antra- cyclines and number of aphereses. At the same time there was Introduction: Autologous peripheral blood stem cells (PBSC) association between the low number of CD34+ cells in the auto- transplantation relies on eff ective preservation of those cells. Cell transplant and the dose of Cph less than 2.5 g/m2. The AutoSCT cryopreservation is a procedure with multiple steps universally was done in 7 of 14 patients with low number of CD34+ cells in the accepted. Nevertheless each center has its own working protocol. autotransplant as there was no any possibility to collect additional At the end of 2010 our center started a new quality program. number of cells according to diff erent reasons. The conditioning Objective: we present our experience, focusing our work in regimens were BEAM and Melphalan 200 mg/m2. The engraft- autologous PBSCs cryopreservation during 2011. We describe our ment of the autotransplant was revealed in all 7 patients on 12-30 protocols and quality controls, including indirect evaluation of (absolute number of granulocyte ≥ 0.5x109/)and 12-100 (platelets engraftment post-PBSC infusion. ≥ 50x109/L) days. But there were much more number of infectious Methods: aphaeresis products are processed in sterile conditions complications (>53%) and number of platelets transfusion (>70%) at the Cryobiology Laboratory (CL) soon after collection, with than in the control group. minimal manipulation. Cell product is distributed to cryopreser- We conclude that if there is no any possibility to treat patient with vation bags and cryoprotectant solution (20% Dimethylsulf- Cph more than 2.5 g/m2 before stem cell collection the combina- oxide (DMSO) in 5% Human Albumin Solution) added to a fi nal tion of G-CSF and plerixafor has to be used.

S520 Lymphocyte biology fl ow cytometry. Assuming that the sample is well mixed this check infers that CD34 stem cell analysis is accurate. A known CD34/ul control is also run at the same time to ensure gating R1474 is accurate. The medicinal mushroom Agaricus blazei Murill activates By limiting the red cell content to 1.75x109 per FACS tube and NF-KB via TLR2 in monocytic cells, activates MDDC and using freshly made lysis buff er this ensures that fl ow cytometry induces expansion of CD56+ HSC but not other progenitor is even more accurate as trucount beads are not depleted by red HSC in vitro cell ghosts. G. Hetland, J. Tangen, A.M. Tryggestad, M. Sand Dyrhaug, G. Kvalheim Oslo University Hospital (Oslo, NO)

Agaricus blazei Murill (AbM) is an edible Brazilian Basidiomycetes Donors and donor selection mushroom from folk medicine with proven immunomodulating and antitumor properties in mice. Methods: Human promonocytic THP-1 cells, peripheral blood R1476 Cd34+ HSC, or monocyte-derived DC (MDDC) (CD1a+, CD14-, In search of a perfect match: a clinical case CD80+)were kept with saline, pure AbM or the AbM-based extract F. Amado, F. Bordalo, S.M. Lopes, S. Ferreira, F. Campilho, A. Campos, AndoSanTM. HSC were cultured for a week in Methocult medium S. Roncon and CFU counted and fenotype determined (fl ow cytometry). IPO Porto (Porto, PT) Results: AbM induced, via TLR2 stimulation, translocation of NF- κ B from cytosol to the nucleus (western blotting)in the THP-1 cells. We present the case of a 12 year old female patient with Fanconi AndoSanTM addition to HSC hampered their general CFU forma- Anemia (FA) diagnosed at age 4, with homozygosity for Fanca tion and expansion dose-dependently, except for CD56+ HSC that gene. The patient was resistant to androgen therapy and trans- increased 2-fold. AndoSanTM induced down-regulation of CD11c, fusion dependent; therefore was a candidate for allogeneic stem up-regulation of CD1a, CD40, CD80, CD83 and CD86, and de novo cell transplant. The search for a related or unrelated HLA compat- expression of CD69 on MDCC. ible donor was unsuccessful. Conclusions: AbM stimulation of THP-1 cells via TLR2 induced This patient was of gipsy ethnicity, with high family consanguinity activation of the nuclear factor NF- κ B transcription factor. After (her parents are fi rst degree cousins). FA was diagnosed in a stimulation with the AbM-based AndoSanTM, MDDC expressed brother and 2 female cousins. Her FA brother died at age 14 after diff erentially markers for adhesion, activation and antigen pre- progression to AML, one cousin died at age 12 after an unrelated sentation, explaining some known antitumor properties of AbM. cord blood unit (CBU) transplant and the other is alive at age 8, in This capacity of AndoSanTM to MDDC diff erentiation fi ts with its search of a compatible donor. general hampering of HSC progenitor cell proliferation. Also, the Being a large family, from 2005 to 2011 we cryopreserved 7 CBU exceptional CD56+ HSC increase agrees with the observed NK cell from her relatives, until an HLA match was fi nally found. We increase in cancer patients taking AbM extract supplementation. decided to propose an allogeneic CBU transplant. The patient was submitted to a conditioning regimen with fl udarabine, cyclosfamide and anti-thymocyte globulin. The CBU was thawed and washed according to the NY Cord Blood Bank protocol prior to infusion. Quality control showed a high cellular con- Graft processing and manipulation tent (2.6x107/Kg nucleated cells, 1.63x105/kg CD34 cells), good clonogenic potencial (17x105 /Kg CFU-GM), excellent viability (95%) and sterility (bacterial, fungal). Syphilis and viral serology R1475 (HBV, HCV, HIV, HTLV, CMV) were not reactive; PCR analysis (CMV in Small sample analysis: reliable results from small volume the CBU and HBV, HCV, HIV in maternal’s peripheral blood), were cord blood segments negative. The c.295 C>T (p.Q99X) mutation was not present. R. Horton (1), D. Gibson (1), R. Davy (1), L. Fry (1), P. Sami (1), The infusion occurred with no immediate complications. Graft C. Leonforte (1), N. Turton (1), M. Marciniak (1), D. White (1), versus host disease (GvHD) prophylaxis was performed with S. Gomez (1), S. Querol (2), A. Madrigal (3) cyclosporine and mycophenolate mofetil. (1)Anthony Nolan (Nottingham, GB); (2)Banc de Sang i Teixits Engraftment occurred for neutrophils at day 15 and for platelet at (Barcelona, ES); (3)Anthony Nolan Research Institute (London, GB) day 21. Presently, at 5 months after transplantation, the patient is in good clinical conditions, with no signs of GVHD. Introduction: cord blood segments typically contain between 50-70ul which means that completing all routine tests upon transplant release or reception can be diffi cult. This is particularly the R1477 case with regards to automated cell counters that can use up to Distribution of human leukocyte antigen match in 350ul for a full diff erential. Our lab has developed a number of patients undergoing allogeneic haematopoietic stem simple methods to ensure that both cell counts and routine fl ow cell transplantation cytometry on small samples is accurate. E. Uzer, M. Cetin, M. Koker Methods: by diluting a small volume of the original sample and University of Erciyes (Kayseri, TR) then scaling up a reliable diff erential can be generated. This allows bi-directional checks of the NC/ul results produced by single Introduction: Analysis of distrubition and HLA match patients platform fl ow cytometry and automated counter. who underwent allogeneic haematopoietic stem cell transplanta- Subsequent fl ow assays and CFU are then performed on the tion (AHSCT) in University of Erciyes, Haematopoietic Stem Cell remaining undiluted sample to reduce potential errors. Also the Transplantation Unit. number of red cells going into the fl ow assays is limited and fresh Patient and methods: Sixty patients who underwent AHCST in lysis buff er is made up just before use in order to minimise lysis University of Erciyes, Faculty of Medicine, Haematopoietic Stem problems as cord blood red cells can be diffi cult to lyse. Cell Transplantation Unit were evaluated retrospectively. Results/Conclusions: dilutions done repeatedly on the Sysmex Results: The diagnosis of patient were 25 acute myeloid leukemia XE-2100 have consistently been shown to be accurate. Up to (AML) (%41,6), 14 acute lymphoblastic leukemia (ALL) (%23,3), a 1:10 dilution can reliably be performed without introducing 2 myelodysplastic sydrome (MDS)(3,3), 6 non Hodgkin lyphoma a large error. As an NC/ul is generated this can then be used to (NHL) (%10), 2 Hodgkin lyphoma (HL) (3,3), 1 sickle cell anemia check the validity of the NC/ul fi gure produced by single platform (SCA) (%1,7), 5 aplastic anemia (AA), 1myelofi brosis, 2 chronic

S521 myelogenous leukemia (CML) (%3,3), 1 chronic lymphocytic R1479 leukemia (CLL) (%1,7), 1 paroxysmal nocturnal hemoglobinuria Cord blood graft composition and its impact on clinical (PNH)(%1,7). The mean age of patients was found 35,3 years outcome after allogeneic stem cell transplantation (min-max:14-59). 28 of patients were female (%46,7) and H. Wikell, M. Uhlin, J. Mattsson, M. Remberger, J. Gertow 32 patients were male (%53,3). Karolinska institutet (Stockholm, SE) Fourty-six patients were underwent AHSCT from 6/6 HLA-A, HLA-B, HLA-DR identical sibling donors, 11 patients were under- Today allogeneic hematopoietic stem cell transplantation (ASCT) went AHSCT from 10/10 HLA- A, HLA-B, HLA-C, HLA-DR, HLA-DQ is used for treatment of hematological malignancies, immuno- identical sibling donors, a patient with AML from 5/6 HLA ideniti- defi ciencies and other non-malignant diseases such as genetic cal sibling donor who had one mismatch in DR locus, a patient and metabolic disorders. Umbilical cord blood (UCB) is a suit- with SCA from 9/10 HLA idenitical sibling donor who had one able source of hematopoietic stem cells when no HLA-matched mismatch in A locus, a patient with CML from 9/10 HLA ideniti- related or unrelated donor is found. The graft composition and its cal sibling donor who had one mismatch in A locus. 40 of donors impact on clinical outcome for PBSC and BM have been studied were male, 20 of donors were female. extensively. Regarding graft composition and clinical outcome Sonuç: Sixty patients who underwent AHCST in University of after cord blood transplantation only a limited amount of data is Erciyes, Faculty of Medicine, Haematopoietic Stem Cell Trans- available. plantation Unit performed AHSCT from all sibling donors. Three We analyzed material from 29 cord blood units used for alloge- patient with 1 mismatch, and fi fty-seven patient with fullmatch. neic hematopoietic stem cell transplantation with multi-color fl ow cytometry. Phenotypical data were correlated with diff erent clinical outcome parameters e.g. survival, graft versus host disease (GVHD), relapse, rejection, viral reactivation, bacteremia etc. The aim was to fi nd cell surface markers that can be used as tools Stem cell sources to predict clinical outcome post transplantation. We found that above-median frequency of CD69+ T cells was associated to patient survival (p=0.025). Additionally, above-median R1478 levels of naïve CD8+ T cells (p=0.028) correlated with survival, and Successful use of plerixafor in hard to mobilize patients: an association between patient survival and above-median levels following high dose therapy all patients developed fast of CD127+ cells B cells (p=0.013) in the graft was found. Moreover, and sustained engraftment with durable clinical responses there was a statistically signifi cant correlation between higher D. Josefsen, G. Hetland, A. Blystad, Y. Fløisand, G. Kvalheim levels of CD94+ total T cells (p=0.0031) as well as CD8+ T cells Oslo University Hospital (Oslo, NO) (p=0.0024) and HSV+VZV reactivation post transplantation. This correlation was also seen for the frequency of CD95+ total T cells Mobilization of autologous hematopoietic stem cells is most fre- (p=0.013), as well as CD4+ (p=0.0067) and CD8+ (p=0.033) T cell quently performed using G-CSF alone. In our clinic we use G-CSF in subsets. Finally, a higher frequency of naïve CD8+ T cells seemed combination with chemotherapy. In 15-20% of the patients insuf- to correlate with aGVHD (p=0.017). fi cient numbers of CD34+ cells are harvested, and are regarded This study highlights the importance of further characterization as poor mobilizers. Furthermore, previous fi ndings among lym- of cellular graft composition and its impact on clinical outcome phoma patients have shown that poor mobilizers have a less after cord blood transplantation. The data obtained may be useful favorable prognosis than good mobilizing patients. Recently, for the post management of patients after CB transplantation. new mobilizing agents including plerixafor, a CXCR4 antagonist, have been developed. We have previously shown that patients mobilized with chemotherapy and G-CSF appearing with low R1480 concentration of CD34+ cells (5-10 cells/μL) and recovered total Assessment of autologous grafts for optimization of periph- white cell count >10x10e9/L, can successfully harvest stem cells eral blood progenitor cells harvest procedures in patients by the addition of plerixafor. In the present study, we have mobi- with haematological malignancies lized and harvested 33 poor-mobilizing patients. Eleven patients A. Pivkova Veljanovska (1), Z. Stojanoski (1), S. Genadieva Stavrik (1), were remobilized with G-CSF and plerixafor, whereas 22 patients L. Chadievski (1), L. Cevreska (1), R. Grubovic (2), S. Useini (2), were given plerixafor upfront in the primary mobilization attempt B. Georgievski (1) in addition to chemotherapy and G-CSF. Furthermore, the 16 lym- (1)University Hematology Hospital (Skopje, MK); (2)Institute for phoma patients were followed up after reinfusion of autologous Transfusion medicine (Skopje, MK) stem cells with regard to short term and long term engraftment as well as relapse and death. Background: Clinical features known as baseline, before start of The day prior to harvest of the 33 patients, the level of leuko- mobilization fail to predict “failure of mobilization” in a substantial cytes was 21,8x10e9 cells/L (median), and the CD34+ concentra- number of patients. The aim of this study was to predict the har- tion was 5,5x10e6/L (median). Following plerixafor injection, the vest timing and successful PBPC collection. Identifi cation of these concentration of CD34+ cells increased to 26,8 x10e6/L (median). factors could be able to predict mobilization failure and can lead The patients were then successfully harvested (median: 3,9x10e6 to use early plerixafor as preemptive agent. CD34+cells/kg) with 1-2 days of apheresis, although one of the Material and patients: A total of 170 autologous recipients were patients only obtained 1.8x10e6 CD34+cells/kg. analyzed. MM 53 pts, NHL 20pts, HD 32 pts and AML in fi rst CR 66 pts. High dose therapy is a curative treatment for lymphomas. There- Mobilization strategies for PBSC for myeloma patients were fore, we focused especially on the clinical outcome on these 16 preformed with CTX 3-5gr/m2 and G-CSF 10mcg/kg or a single patients. Following high dose therapy, time to short term engraft- agent G-CSF 10mcg/kg in a 5 days duration. In patients with ment, defi ned by neutrophils >0.5x10e9/L and thrombocytes NHL and HD aphaeresis was preformed mainly with single agent >20x10e9/L were 11 days (median) and 20 days (median). More- G-CSF or after HDT with DHAP (for HD patients) or DexaBEAM for over, in contrast to previous fi ndings we have observed durable NHL patients. High dose VP-16 2000 mg/m2 plus G-CSF 10mcg/kg responses with relapse free survival of 77%, and overall survival of was also used as mobilizing regimen in AML pts. Criteria used to 93%, with a median observation time of 14 months. start apheresis were PB CD34 value over 20mmc. Logistic regres- In conclusion, our fi ndings show that addition of plerixafor in sion was utilized to determine predictive factors for stem cell hard to mobilize patients allow them to proceed to high dose mobilization. therapy. Moreover, the additional costs related to the use of Results: A failure of CD34 cell mobilization in PB was registered in plerixafor can be justifi ed since lymphoma patients obtain similar 6% of patients; a failure to collect cells for 1 high dose treatment event free and overall survival as those patients that were good in 11.5% of all patients, a failure to collect in 2 high dose courses mobilizers. was registered in 33%. A successful CD34 mobilization was not

S522 predicted by any factors. The harvest of CD34>2.0x10(6)/kg was achieved by Granulocyte-Colony Stimulating Factor (G-CSF) in predicted in univariante analysis by male gender (p=0, 01), body combination with chemotherapy. In non-hematological disorders weight >90kg (p=0.003), baseline WBC count (p=0.01), percent of the mobilizing chemotherapy was cyclophosphamide 4g/m2. In MNC at apheresis day (p=0.01), number of apheresis procedures patients with hematological malignancies mobilization chemo- (p=0.01) and number >2 lines of induction CT (p=0.01). Multivari- therapy was cyclophosphamide 4g/m2, 7g/m2, Ara-C 8g/m2, ate analysis revealed harvest failure in patients with BM residual IGEV, DHAP, IELSG32, HD MTX-AraC. infi ltration (p=0.003), hypocellular BM (P=0.001) and Plt count Results: Time of mobilization, the day expected for collection, the below 120x10(9)/L (p=0.0001). The use of drugs such as Thalido- suggested optimal day to start chemotherapy are listed in Table 1, mide and RT in did not have impact on apheresis. according to chemotherapy. Conclusion: factors that are able to predict suffi cient mobilization Conclusion: A continuous data record and analysis allows identifi - are not easily identifi able at baseline. The criteria to asses mobili- cation of specifi c days in which start chemotherapy, so scheduling zation failure should still be analyzed so we are able to avoid an PBSC collection in working days and decreasing additional costs inappropriate use of expensive agents like plerixafor early “on related to engagement of Collection Facility, Flow Cytometry demand” basis. laboratory and Cryopreservation Facility on holidays.

R1481 R1482 Optimizing PBSC collection in working days: results of Analysis of mobilization and remobilization regimens 121 patients undergoing chemotherapy plus G-CSF to achieve suffi cient stem cell yield for autologous S. Gattillo, M. Coppola, L. Malabarba, S. Malato, L. Barzizza, transplantation. Improve mobilization strategy: single S. Marktel, M. Bruno Ventre, M. Marcatti, M. Zambelli, C. Parisi, centre experience (859) R. Milani, F. Ciceri, L. Bellio P. Ganeva, G. Arnaudov, M. Mincheff , I. Tonev, C. Botev, J. Jechev, San Raff aele Scientifi c Institute (Milan, IT) M. Guenova, A. Mihova, N. Stojanov, P. Todorova, G. Mihaylov, S. Toshkov Objectives: Autologous Peripheral Blood Stem Cells (ASCT) trans- Hospital for Hematological diseases (Sofi a, BG) plantation is a standard of care for some hematological disorders. Stem cell collection for autologous use is increasingly targeted at Background aims: The aim for this study was to analyze the doses exceeding the need for one single procedure, in order to factors aff ecting the autologous stem cell mobilization (ASCT) in allow programs of sequential intensive chemotherapy (HDS). our BMT Unit CIC 859 (Specialised Hospital for Hematological Dis- An accurate scheduling of mobilizing chemotherapy is a relevant eases - Sofi a Bulgaria). for the period between January 2009 to pre-requisite to maximise the timing of stem cell collection within September 2012. Successful mobilisation was defi ned as collec- the weekly working days. We analysed apheresis timing according tion of more than 2x10 6 CD34+ cells/kg. The eff ect of patient age, to starting chemotherapy days to allow improvement in organiza- diagnosis, type and number of chemotherapy regimens, diseases tion-related costs. invasion of bone marrow, type of mobilizing protocols on the Methods: From January 2011 to November 2012, we managed collection effi ciency of PBSC was investigated. 121 patients, 113 with hematological malignancies and 8 with Methods: We analyzed records of 284 patients underwent stem cell non-hematological diseases. Ten patients resulted poor mobiliz- mobilization for ASCT. From all patients 134 ( 47.2 % ) were with ers; we evaluated 111 patients who were eff ectively harvested by multiple myeloma, 76 (26.8 %) with non- Hodgkin’s lymphoma, leukoapheresis. In all patients, mobilization of CD34+ has been 74 (26.1 %) were with Hodgkin’s lymphoma. A total of 138 (48.6 %) patients received only granulocyte- colony- stimulating factor (G-CSF) and 146 (51.4 %) received G-CSF plus chemotherapy (G-CSF+C) for the initial mobilization. After the mobilization peripheral blood CD 34+ cells were analysed by fl ow cytometry. Results: The median CD 34+ cell yield was higher in G-CSF+ C group than in G-CSF group. The failure rates were : from all 284 patients 45 (15.18% ) were bad mobilizers; 21 (47.4 % ) in G -CSF group and 23 (52.27 %) in G-CSF +C group respectively. 26 (59 %) of 45 “bad mobilized” patients underwent remobilization using only G-CSF a/C+ G-CSF or G-CSF + Plerixafor. 17 patients (65 %) received Plerixafor for remobilization. In 11 of these patients Plerixafor was used for their second mobilization, in 6 - for third and next remobilization; Twenty three ( 88.4 %) of all remobilized patients achieved more than 2 x 10 6 CD34+ cell/kg and 3 (11.6 %) failed to pool suffi cient number of stem cells from both collections. Patients receiving G-CSF + Plerixafor had lowest failure rates. Only 1 patients had unsuffi cient remobilization wth Plerixafor. Conclusions: Early identifi cation of patients who are diffi cult to mobilize is important as these patients will be candidate for additional mobilization. Using G-CSF + Plerixafor increases mobilization yield and improve effi ciency of this treatment by cost and time saving.

R1483 Graft content and engraftment relationship in allogeneic bone marrow transplantation S. Civriz Bozdag (1), P. Topcuoglu (2), M. Bay (2), E. Ayyildiz (2), O. Arslan (2), M. Beksac (2), G. Gurman (2), O. Ilhan (2) (1)Ankara Oncology Hospital (Ankara, TR); (2)Ankara University Hematology Department (Ankara, TR)

Today peripheral blood is still the mostly preferred stem cell source in transplantation, bone marrow still remains an option

S523 due to its less chronic GVHD rates and similar survival. In this In summary, cHCB HSCT is feasible in children and potentially study, we aimed to analyze the relation between infused stem combines benefi ts of both stem cell sources: rapid Haplo-derived cell amount and engraftment in peripheral blood/bone marrow neutrophil recovery shortens aplasia, and patients may benefi t transplantation. from the superior adaptive immune system provided by the uCB We included 102 allogeneic bone marrow transplantation (BMT) graft. Moreover, cHCB potentially enables the use of uCBs with patients and 273 allogeneic peripheral blood transplantation marginal cell counts, and carries the possibility of post-transplant (PBT) patients in Ankara University BMT Unit. Two patients who adaptive immunotherapy. The risk of both GvHD and rejection had transplantation from unrelated donors and 15 patients whose remains to be determined. Further research is therefore necessary donors received granulocyte colony stimulating factor before to evaluate the role of cHCB transplantation in children. Never- transplantation were excluded from the bone marrow transplan- theless, this is a promising option for patients without a matched tation arm.All patients had transplantation from HLA identical sibling or unrelated donor who require HSCT urgently. relatives; in bone marrow group; median age was 26 (16-57), Male/ Female was 53/32. Median amount of collected stem cell product was 1155 (392-1928) cc, ertyhrocyte depleted stem cell prod- R1485 ucts for ABO major missmatched patients were 143 (78-178) cc. Mobilization of peripheral blood stem cells : Tunisian Median collected CD34+ cell number was 2,4 (0,3-7,5) x 10e6/kg, experience median nucleated cell number was 2,3 (0,5-5,2) x 10e8/kg and S. Zriba (1), H. Ghédira (1), K. Kacem (2), N. Sakka (2), R. Ben lakhal median mononuclear cell number was 0,59 (0,3-3,02) x 10e8/kg. (2), R. Mansouri (2), M. Zarrouk (2), M. Maamer (3), Y. Ben abdennebi In PBT group; median age was 36 (164), M/F 111/162, median CD34 (2), H. Ben neji (2), R. Jeddi (2), Z. Belhadj ali (2), H. Ben abid (2), was 5,7 (2-15). In BMT group; the median posttransplantation day F. M’sadek (1), S. Hmida (3), B. Meddeb (2) for neutrophil and platelet engraftment was 16 (11-28) and 22 (1)Mlitary Hospital (Tunis, TN); (2)Aziza Othmana Hospital (9-69) respectively. Four patients died in the aplasia period due (Tunis, TN); (3)National Center For Blood Transfusion (Tunis, TN) to sepsis.One patient had primary engraftment failure and had one more stem cell infusion, since then he has been followed in Background: High-dose therapy with peripheral blood stem cells complete remission. Patients were grouped according to infused support is a standard of care in patients with lymphoma particu- CD34 and TNC numbers. Neutrophil and platelet engraftment larly in refractory and relapsed disease. Minimal dose of 2 x106 days were statistically nonsignifi cant between groups who had CD34+ cells/kg collected is required for this procedure. infused CD34+ cells lower and higher than 2x 10e6/kg (p: 0.334, Methods: Data about mobilization of 105 patients with lymphoma p: 0.080, respectively). But both neutrophile and platelet engraft- treated in two Tunisian haematology departments between 2005 ment days were lower in patients transplanted with TNC numbers and 2011 were collected. higher than 2x 10e8/kg (p<0.05 and p<0.05, respectively). In PBT Results: 105 patients (59 males and 46 females) with Hodgkin’s group; median posttransplantation day for neutrophil and plate- lymphoma (HL) (n=50) and non-Hodgkin’s lymphoma (NHL) let engraftment was 15 (8-47) and 13 (5-43) respectively. Thirteen (n=55) were mobilized. The median age was 29 years (range, of 273 patients (4%) and thirty of 273 (10%) patients could not 14-61) and 84% of patients were aged less than or equal to have neutrophile and platelet engraftment respectively. The yield 50 years. 75% of patients had stage III-IV disease. Initial bone mar- of bone marrow graft is one of the major factors that has infl u- row infi ltration was present in 18%. 18% of patients experienced ence on engraftment. Similar to previous studies, we showed in more than 2 therapeutic regimen and 34% more than 6 cycles of our study that, TNC numbers of the graft seems to be superior to chemotherapy (median was 4). Seven patients had received previ- CD34+ cell number to predict the engraftment kinetics. ous radiotherapy of residual sites. The mobilization strategy was a combination of chemotherapy and G-CSF: Filgrastim (n=72) and Lenograstim (n=33). Diff erent dosages of G-CSF were used: R1484 5 mcg/kg/d in 31 patients, 10 mcg/kg/d in 5 patients and 5 mcg/ Co-infusion of haploidentical T-cell depleted stem cells kg/d then 10 mcg/kg/d from day 10 in 69 patients. The median shortens the interval to neutrophil recovery after cord number of leukapheresis was 1 (range, 1-4) with a median time of blood transplantation: three paediatric patients 14 days (range, 10-28) after chemomobilization. 60% of patients M. Mynarek (1), B. Maecker-Kolhoff (1), R. Beier (1), M.G. Sauer (1), had only one apheresis. A median of 6.06 x106/kg CD34+ cells were C. Klein (2), K.W. Sykora (1) collected (range, 1.1-30). 12 patients failed to collect the minimal (1)Hannover Medical School (Hannover, DE); (2)Ludwig-Maximilians dose of 2 x106 CD34+ cells/kg. The following data were identifi ed University (Munich, DE) as determinant for failure of collection: age > 50 years (p=0.011), Hemoglobin rate < 10 g/dl (p=0.012) and initial bone marrow Haploidentical transplantations are limited by delayed immune involvement (p=0.002). The collection yield was signifi cantly reconstitution and unrelated Cord blood (uCB)-transplantations higher in HL patients (p=0.009), in younger patients ≤ 50 years by prolonged neutrophil and platelet engraftment. A combined (p=0.0001), and with dose of G-CSF (5 mcg/kg/d then 10 mcg/kg/d Haplo- and uCB- (cHCB) transplantation may be an option to over- from day 10) p=0.039. Age less than 40 years was a predictive come these limitations in patients who do not have a matched factor for successful collection with one apheresis (p=0.035). sibling donor and cannot wait for a matched unrelated donor. Conclusion: In our country we proposed dose escalation of G-CSF We present the results of cHCB HSCT in three children after con- (5 mcg/kg/d then 10 mcg/kg/d from day 10) for patients having ditioning with fl udarabine, treosulfan, thiotepa, and ATG or ALG. predictive factors of failure at initial mobilization. We reserved the Cyclosporin A and mycophenolate mofetil were used as pro- use of plerixafor for patients with proven failure. phylaxis for graft versus host disease (GvHD). All received either 5/6 or 6/6 HLA-matched uCB grafts with a cell dose of 0.66x108 to 1.23x108 nucleated cells/kg. Two of the patients received simulta- R1486 neous (±1day) paternal CD34-positive selected peripheral blood Cellular graft composition infl uences on engraftment, GvHD stem cells (PBSCs) and one patient received maternal CD3/CD19- reaction in transplantations from match unrelated donor depleted PBSCs with a cell dose of 18.8x106 to 36.33x106 CD34- Y. Mareika, T. Shman, S. Sharapova, A. Nevmerzhitskaya, positive cells/kg. All of the patients had leukocyte engraftment A. Alekseichik, N. Minakovskaya between day +10 and day +11, predominantly derived from the Belarusian Research Center for Pediatric Oncology, Hematology and parental donors. In two patients, chimerism converted to 100% Immunology (Minsk, BY) uCB between day +28 and day +100. The third patient rejected both the Haplo and the uCB-graft and had autologous recovery Cellular populations of the graft have been shown to infl uence on on day +120. None of the patients developed acute or chronic hematopoietic recovery, infection and graft versus host disease GvHD. All patients are alive and well with 9.5 months to 4.5 years (GvHD) complications and overall survival. The subject of our follow-up after transplantation. study was to assess the impact of diff erent cell types of graft on

S524 engraftment, acute and chronic GvHD reactions, early immuno- mononuclear cell dose per kg was 3.68 x107, while CD 34+ cell dose logical recovery. In the study group were included 20 patients per kg was 1.58 x105. Conditioning regimens used were busulfex, (aged from 1 to 19) who received transplants from matched unre- fl udarabine, and rabbit ATG. Cyclosporine was used as GVHD pro- lated donors (MUD). Bone marrow (n=11) and mobilized PSCT phylaxis. She had neutrophil recovery on day +14. Besides mild (n=9) were sources of the grafts. Quantity of CD34+, CD34+CD38- transient mucositis, there were no any other serious complications cells, T-cells and T-cell subpopulations, B-, NK-cells were analyzed but still anemia. Major BO incompatibility caused her red cells by fl ow cytometry. Speed of leukocytes and neutrophils engraft- slow recovery. That was solved later by 2 cycles of plasmapher- ment correlated signifi cantly with total transplanted nucleated esis done on 24, 25 July 2012. Her high anti-B titer has decreased cells as well as CD34+ ones. However, the strongest association to negative. Her baseline hemoglobin rose to over 11.5 g/dL. was found between dose of CD34+CD38- stem cells and engraft- Her blood group changed from O+ to B+. Her β globin gene ment of leukocytes (Rs=-0,65 Er=0,002) and neutrophils (Rs=-0,56 analysis turned to hemoglobin E trait. Post-transplant studies Er=0,009). No correlation was found between doses of studied showed full donor engraftment. To date it is almost 1 year follow- cells populations and platelets engraftment. up, she is doing well, no transfusion-requirement, no any GVHD. Patients who developed acute GvHD reaction staged from 2 to 4 received signifi cantly higher doses of total nucleated cells, CD34+ and CD34+CD38- stem cells, CD19+ B-cells, CD3+CD56+ R1488 EKT-cells and CD4+CD25+CD127- Treg cells. Particularly, dose of Change in haematopoietic stem cell count in peripheral CD34+CD38- cells in the group of patients with acute GvHD occur- blood following blood donation rences was 2,0 (1,0-3,9)X106 /kg whereas in the group of patients C. Pala, H. Mumcuoglu, F. Kurnaz, S. Sivgin, L. Kaynar, M. Keklik, without acute GvHD it was 0,49 (0,3-1,0)X106 /kg (Er=0,03). Early G. Akyol, I. Altun, Y. Koker, B. Eser immunological recovery was also associated with doses of trans- Erciyes University (Kayseri, TR) planted cell subpopulations. Absolute counts of CD3+ T-cell, CD3+CD8+ T-cytotoxic cells, CD3+HLA-DR+ activated T-cell at day We aimed to investigate the change in the number of stem cells +30 were strongly related with total dose of transplanted nucle- and white cells in the early period following blood donation. ated cells as well as CD34+ cells. However, CD19+ cells absolute 22 male (71%) and 9 female (29%), 31 volunteers in total were count was infl uenced by CD34+CD38- stem cells dose. included in the study. 450 ml of whole blood were collected from Grafts of patients who developed chronic GvHD reaction had each of the volunteers for the donation. Complete blood counts signifi cantly higher percentage of CD3+CD4+ T-helper cells, were performed on the volunteers before and at 6 and 24 hours CD3+CD8+ T-cytotoxic cells, CD3-CD56+ NK-cells and CD19+ after the donation and CD34+ cell counts per ml of peripheral B-cells. Doses of total transplanted nucleated cells and CD34+ blood were measured by fl ow cytometry technique. cells were not associated with chronic GvHD reaction. There was a statistically signifi cant increase in the number of In conclusion, in MUD transplantations diff erent cells subpopula- CD34+ cells in the peripheral blood at 6 hours following blood tions of graft have impact on speed of engraftment, immunologi- donation (p <0.001). At 24 hours, however, there was a statistical recovery and occurrence of acute and chronic GvHD. cally signifi cant decrease in the number of CD34+ cells, compared to 6 hours (p<0.001). There was a statistically signifi cant increase in the number of white cells in the peripheral blood at 6 hours R1487 following blood donation (p <0.001). At 24 hours, there was a Successful stem cell transplantation for β-thalassemia child decrease in the number of white cells, which was statistically sig- using cord blood from newborn sibling achieved by in-vitro nifi cant compared to 6 hours(p <0.001). When the diff erence in fertilization, preimplantation genetic diagnosis and human CD34+ cell and white cell counts before blood donation and at leukocyte antigen matching methods: the fi rst report from 24 hours after blood donation were compared, the results were East Asia not statistically signifi cant. P. Vanichsetakul (1), S. Manipalviratn (2), K. Tiewsiri (2), J. Fongsarun A transient increase in the number of CD34+ cells in the periph- (3), K.I. Papadopoulos (3) eral blood after blood donation was demonstrated, with a decline (1)Wattanosoth Hospital, Bangkok Hospital Group (Bangkok, TH); in CD34+ cell counts back to levels prior to donation at 24 hours. (2)SUPERIOR A. R. T. (Bangkok, TH); (3)THAI STEMLIFE (Bangkok, TH)

Β-thalassemia diseases are common in Southeast Asia. Alloge- R1489 neic stem cell transplantation is worldwide acceptance as cura- T-cell replete non-myeloablative allogeneic peripheral blood tive therapy. When aff ected child has no HLA-matched siblings or stem cell transplantation from HLA haploidentical family unrelated donors, umbilical cord blood (UCB) transplant from HLA- donors after preparation using busulfan, fl udarabine and matched newly born sibling may be considered, if feasible. The fi rst anti-thymocyte globulin ever event from East Asia reported here is about allograft trans- C.S. Kim (1), I. Kim (2), H.G. Yi (1), Y.H. Park (1), S.S. Yoon (2), plantation performed in a transfusion-dependent β-thalassemia/ S. Park (2) hemoglobin E, 5-year-old Thai girl, using UCB stem cells from her (1)Inha University Hospital (Incheon, KR); (2)Seoul National HLA-matched younger brother who was achieved by in-vitro fertil- University Hospital (Seoul, KR) ization (IVF), preimplantation genetic diagnosis (PGD), and human leukocyte antigen (HLA) matching methods. Upon the full discus- In July 2010-June 2012, allogeneic peripheral blood stem cell sion and the parents’ desire to have a non-aff ected; new baby, the transplantation from HLA haploidentical family donor (haplo- IVF; PGD; and HLA matching methods were done successfully by PBSCT) was attempted for 7 patients (4 males, 3 females) at the Superior A.R.T. center. After the mother’s uneventful, complete median age of 57 (range 14-63) diagnosed as acute lymphoid leu- pregnancy, the donor was delivered by C-section on 21 January kemia (ALL) with positive Philadelphia chromosome (n=3, 2 in CR1, 2012, as a full-term healthy male baby. UCB was collected there- 1 in CR2), acute myeloid leukemia secondary to myelodysplasia in after by team of Thai StemLife Company. The patient had blood CR1 (n=1), ALL in CR2 preceded by peripheral T cell lymphoma group O+ but the donor had group B+. His β globin gene analysis (n=1) and severe aplastic anemia (n=2). Non-myeloablative condi- was confi rmed to be heterozygous codon 26 (Hb E) G->A. His UCB tioning regimen Bu-Flu-ATG consisted of busulfan 3.2mg/kg IV on HLA type was also confi rmed as completely matched. As UCB stem days -7 to -6, fl udarabine 30mg/m sq IV on days -7 to -2 and rabbit cell dose being estimated beforehand and considered suffi cient anti-thymocyte globulin 3mg/kg IV on days -3 to -1. Cyclosporine for transplant, that cryopreserved UCB was thawed and infused 3mg/kg/day and methotrexate 15mg/m sq on day 1 and 10mg/m into the patient on 30 April 2012. The patient’s weight was 17.9 kg. sq on days 3, 6 and 11 were given for graft versus host disease Prior to transplant, she had received 60 times of leuko-depleted (GVHD) prophylaxis. Donor blood stem cells were mobilised with packed red cell transfusion with regular use of oral deferasirox. G-CSF at 10ug/kg daily from day -3 to day 0 and continued until the She was estimated as Pesaro class II. Regarding UCB infusion, end of the harvest. Blood stem cells were collected by apheresis

S525 for 1-3 days and given without T cell depletion. Medians of been made to this scheme with the substitution of drugs (eg car- 6.24x10(6) CD34+cells/kg (range 4.22-10.5), 11.2x10(8) NMCs/kg mustine by Thiotepa (TEAM) fotemustine (EGF)) Chemosensitivity (range 5-16.5) and 3.8x10(8) CD3+cells/kg (range 2.2-12) were successfully in transplantation (70-80% complete responses). In infused on day 0, days 0-1 or days 0-2. Engraftment was achieved our study was substituted for cisplatin , taking into account the in all patients on the median of day 13 (range 10-24) with neu- antitumor activity of this agent in lymphomas, with satisfactory trophil count>500/uL. Platelet count>20,000/uL was seen on the results in 85% overall response after transplantation. median of day 28 (range 13-293). Complete donor chimerism was documented for all on day 28. Prophylactic trimethoprim- sulfamethoxazole was started after engraftment and continued R1491 until day 180 with cyclosporine. Veno-occlusive disease was Sequential allogeneic stem cell transplantation in high absent. Acute GVHD developed in 3 and one died of gut GVHD risk acute myeloid leukaemia and myelodysplastic with massive GI bleeding and septicemia on day 147. Febrile syndrome episodes developed in 6 occasions, of which S. aureus (day 9), A Zavrelova, J. Radocha, M. Lanska, P. Zak P. aeruginosa (day 119) and A. baumannii (day 142) were bacterio- 4th Department of Internal Medicine - hematology logically proven. S. aureus caused early death. Reactivation of CMV (Hradec Králové, CZ) occurred in 3 requiring preemptive ganciclovir, BKV in 2, herpes simplex in 1, chicken pox in 1, and EBV in 1 with lymphoprolifera- Aim: To evaluate this strategy in high risk acute myeloid leukemia tion requiring rituximab. Chronic GVHD was absent except for one (AML) and myelodysplastic syndrome (MDS). that developed in continuation from acute GVHD. Two patients We treated 17 patients with AML or MDS. This cohort consisted of died on days 33 and 147 by treatment related complications. nine patients who did not reach complete remission, 5 patients As of this writing, 5 patients are alive disease free at a median of who reached complete remission but remained with high burden 512 days (range 210-847) post haplo-PBSCT. Bu-Flu-ATG, parti- of residual disease and 3 patients up front transplanted for myelo- cularly ATG, appears to play an important role in overriding HLA dysplatic syndrome with high blast count. Median age was 54 barrier against T cell replete haplo-PBSCT. years (32-65years). Median count of blasts in bone marrow before sequential transplantation was 9% (1-76%). 6 patients were transplanted with HLA identical sibling, 8 patients were transplanted with matched unrelated donor (10/10) and 3 patients had unrelated donor with one mismatch (9/10). Flag-Ida chemotherapy Conditioning therapies (CHT) (n=4), 3+7 CHT (n=7) or CHT based on monotherapy with Cytarabin was used as a cytoreduction. Between cytoreductive CHT and preparative regimen was instituted rest period with R1490 median of 5 days (2-14 days). After these days of rest prepara- Conditioning regimen with cisplatin (PEAM) versus BEAM tive regimen consisted of Fludarabin/Busulfan was initiated. Total and rescue with autologous haematopoietic progenitor dose o Busulfan was 8mg/kg n=6), 12mg/kg(n=6) nad 16mg/kg cells in lymphoma (n=5). Thymoglobulin (total dose 7,5mg/kg) was used in case of L. Valero, H. Zurita, R. Espinoza, L. Rivera, C. Arroyo, J. Labardini unrelated donor in days -3 to -1. Instituto Nacional de Cancerologia (Mexico City, MX) Results: Estimated overall survival at 2 years was 67,6%. Whole regimen was tolerated very well even with the highest dose High-dose chemotherapy and autologous hematopoietic stem of Busulfan. Transplant related mortality at 100 days was cell transplantation is the treatment for patients with relapsed 6% (1 patient with insufi tient engraftment died due to infection). lymphoma with chemosensitive disease and the results are 5 patients relapsed after transplantation, 1 patient progressed better when it is compared with the standard dose of various after transplantation. From these patients just 1 patient did not chemotherapy regimens, many factors infl uence the outcome die due to relapse and was succesfully rescued with chemotherapy after transplantation. and donor lymphocyte infusion. In order to improve performance, various schemes have been Conclusion: Sequential transplantation is our method of choice used conditioning with the inclusion of new drugs to, but has in high risk AML and MDS. We demonstrate very low transplant not shown a clear superiority compared to the standard regimen related mortality and promising long term survival of this cohort is BEAM. of high risk patients. Objective: This study aims to compare the short-term results in terms of chemosensitivity, disease-free survival and Global conditioning with a scheme in which we replaced carmustine cisplatin R1492 (regimen PEAM) and compared with a similar group of patients Prognostic factors and outcomes in allogeneic haematopoi- BEAM received standard schema. etic cell transplant for patients aged 50 or more Material and Methods: Was performed comparing two groups P. Iniesta, C. Castilla-Llorente, I. Heras, C. Martínez-Millán, V. López, with subtypes of lymphomas in which 20 patients were admin- F. de Arriba, J. Cerezo-Manchado, V. Roldán, M. Suárez, V. Vicente istered the packaging scheme based on cisplatin 100mg/m2 day Hospital Universitario Morales Meseguer (Murcia, ES) -4 SC, SC 1500mg/m2 Etoposide days -4, -3, cytarabine 800 mg / SC m2 day -4, -3, -2 and 140mg/m2 SC Melphalan days -4 (PEAM) We performed a retrospective study of patients aged 50 or more and 18 patients received conditioning scheme based on day -4 who underwent allogeneic hematopoietic cell transplant (HCT) 300mg/m2 Carmustine, Etoposide SC 1500mg/m2 days -4, -3, Cyta- from 1997 to 2011 in our Unit. All clinical outcomes were ana- rabine 800 mg / m2 SC days -4, -3, -2 and 140mg/m2 SC Melphalan lyzed. In order to identify risk and prognostic factors associated or days -4 (BEAM). not to age, study variables were analyzed for the whole group and Results: A total of 38 patients who received conditioning scheme in two diff erent age groups: 50 to 59 and 60 or more. Statistical for Lymphoid Neoplasms ASCT in 20 patients and 18 patients analysis was performed with SPSS.17.0 software. PEAM BEAM scheme. Hematological recovery in neutrophils in Results: 70 patients were included, median age 58 years (range PEAM group was 11 days (10 to 13d) and for BEAM was 12 days (10 50-68). 81% (n=57) received reduced intensity conditioning regi- to 20 d). The response measured by PET CT at 2 months of ASCT men, and 19% (n=13, all <60 years) myeloablative conditioning was 75% for group PEAM vs. 44% for BEAM group. Disease-free regimens. 79% of patients (n=55) received HCT from related survival and Global at 1 year was 70% and 90% for group PEAM donors and 93% (n=65) were HLA-identical. Graft source was and 33% and 88% for the BEAM. peripheral blood in 93% of patients (n=65). Global incidence of Conclusion: The BEAM scheme remains the standard scheme of acute graft versus host disease (GVHD) was 40% (n=28) and of conditioning for Hodgkin and non-Hodgkin enough with toler- chronic GVHD of 53% (n=37). Other than the intensity of the condi- able toxicity and antitumor activity, several modifi cations have tioning regimen, there were no statistically signifi cant diff erences

S526 on graft source, GVHD prophylaxis, and acute and chronic GVHD R1494 incidence and other clinical complications among the two age Haploidentical unmanipulated bone marrow transplantation groups. For a median follow up of 21 months, overall survival (OS), and post-transplantation cyclophosphamide for patient with disease free survival (DFS) and transplant related mortality (TRM) high risk of acute non-lymphoid leukaemia were 53%, 43% y 32% respectively. On the univariate analysis HLA G. Messina, G. Console, T. Moscato, R. Fedele, M. Martino, L. Russo, disparity, unrelated donor (URD) and the presence of acute GVHD M. Cuzzola, A. Pontari, I. Callea, E. Spiniello, G. Irrera were adverse prognostic factors for OS (p=0.007;0.006 and 0.025 AO Bianchi Melacrino Morelli (Reggio Calabria, IT) respectively). The presence of chronic GVHD was a statistically signifi cant protector factor for OS and DFS (p=0.000). In the multivari- Introduction: In the last 15 years for patients aff ected by Acute ate analysis, only the development of GVHD maintain signifi cance Non Lymphoid Leukemia (ANLL) with poor prognostic factors and for OS, being acute GVHD an adverse prognostic factor (RR: 2.13, lacking of HLA-matched donor, we used therapeutic strategies in IC 95% 1.01-4.45, p=0.044) and chronic GVHD a protector factor accordance with the group of Perugia and with Chinese group. (HR:0.25, IC: 95% 0.11-0.56, p=0.001). In our cohort, the disease In our opinion in both instances, the experience was extremely status and HCT-CI of 3 or more were not statistically signifi cant disappointing due to the high toxicity and mortality. Recently, for OS, TRM or DFS, neither for the whole group nor for the 2 age the Baltimore group used a conditioning regimen with moder- group separately. ate intensity, infusion of bone marrow unmanipulated and Cyclo- Conclusions: These results suggest that the unrelated donor graft phosphamide as the primary drug for the prophylaxis of Graft source and HLA disparity, as long as the GVHD development are versus Host Disease (GVHD). On the basis of this experience and prognostic factors for elderly patients undergoing HSCT, whereas intensifying the original schedule of conditioning regimen, we the age, conditioning regimen and higher HCT-CI were not treated 8 patients (pts). signifi cant. Patients and Methods: We utilized a conditioning regimen with Thiotepa 10 mg/bw instead of Cyclophosphamide, Fludara- bine (150 mg/sm) and Total Body Irradiation 2 Gy in 5 male and R1493 3 females with median age of 35 yy (r.6-60). GVHD prophylaxis Allogeneic stem-cell transplantation after a myeloablative was Cyclophosphamide 50 mg/bw on days +3 and +4 and then reduced-intensity conditioning regimen with fl udarabine, Tacrolimus and Mycophenolate Mofetil. All pts received Granulo- busulfan and antithymocyte globulin in patients older cyte-Colony Stimulating Factor. At time of transplant n. 6 pts were than 45 years in I Complete Remission (CR), n. 2 in II CR, but all were stratifi ed in D. Martineau, R. Tabrizi, A. Pigneux, K. Bouabdallah, T. Leguay, high risk for cytogenetic or biological features at diagnosis. Two M. Dilhuydy, A. Lascaux, C. Duclos, G. Marit, N. Milpied pts already underwent an autologous transplant. A median num- Service d’Hématologie (Pessac, FR) ber of marrow mononucleated cells of 360x10E8/kg (r.246-795) were infused. Allogeneic stem-cell transplantation after a non-myeloablative Results: All pts, but one, engrafted. This patient engrafted after conditioning regimen in older patients is commonly associated autologous peripheral stem cell rescue. The overall median time with a reduced incidence of early non-relapse mortality (NRM). for Absolute Neutrophil Count (>0.5x10E9/l) was 18 days (r.14-25) However, the risk of relapse remains signifi cant and has to and 27 days (r.17-65) for platelet count (>30x10E9/l). No signifi - be reduced to improve the overall outcome. The recent devel- cant complications occurred during the cytopenia except for an opment of myeloablative and reduced-intensity condition- episode of hemorrhagic cystitis. We documented 5 cases of cyto- ing regimes may off er such an opportunity without increasing megalovirus reactivation. The full chimerism was reached in all NRM, but a thorough evaluation is warranted to examine this pts within the hundredth day. At 100 day acute GVHD was grade hypothesis. Here, we report our experience of patients older than II for 12,5% of cases. We observed neither acute GVHD grade 45 years transplanted with a myeloablative reduced-intensity III-IV nor chronic GVHD. At median follow-up of 7 months (r. 5-26), conditioning regimen combining fl udarabine 150 mg/m2 (30 mg/ Transplant Related Mortality was 12,5%, while Progression-Free m2/d from d-6 to d-2), IV Busulfan 130 mg/m2/d from d-6 to d-3 Survival and Overall Survival were 87,5%. and antithymocyte globulin (Thymoglobulines®: 2.5 mg/kg/d Conclusion: The unmanipulated bone marrow transplanta- at d-2, d-1). tion from haploidentical family donor and post-transplantation Twenty-four patients transplanted between Jul. 2011 and Oct. Cyclophosphamide provide very encouraging results in terms of 2012 are reported. Their median age was 57 years (45-64). There engraftment rate, GVHD incidence and survival and represents a were 13 males. Diseases and status at transplant were AML, n=9 feasible and valid alternative for pts with high risk of ANLL, lacking (6 CR1, 3 CR2); MDS, n=4 (3 untreated, 1 refractory); CLL, n=3 (3 ≥ an HLA identical sibling. PR2); myeloma, n=2 (1 ≥ PR2, 1 refractory), NHL, n=1 (CR2); Hodg- kin’s disease, n=1 (≥CR2); myelofi brosis, n=4 (3 untreated, 1 PR1). Donors were matched related (13) or matched unrelated 10/10 at R1495 the allele level (12). Source of stem cells were peripheral blood Planned second (PLAS) allogeneic transplantation for (22) or bone marrow (2). The median number of CD34+ cells was refractory acute leukaemia 4.7 x 106/kg (1.3-11). Prophylaxis of GVHD were CsA+MTX (21), CsA M.Y,. Shapira, I.B. Resnick, S. Grisariu, B. Avni, L. Dray, S. Samuel, R. Or (2) or CsA+MMF (1). All patients but 1 engrafted. The patient who Hadassah – Hebrew University Medical Center (Jerusalem, IL) did not engraft had an untreated RAEB-2 before transplantation with a matched unrelated donor and is still alive at 4 months. The Refractory acute leukemia (AL) occurs in a signifi cant percentage median times to reach ANC > 500/mm3, > 1000/mm3, platelets of the AL patients and presents a therapeutic challenge. Allo- > 20,000/ mm3 and > 50,000/ mm3 were 15 days (11-35), 19 days geneic stem cell transplantation (allo-SCT) is the only curative (12-35), 15 days (0-233) and 20 days (11-396), respectively. The option for these patients. Although many of the patients with incidences of acute GVHD II-IV, acute GVHD III-IV, and extensive refractory AL that undergo myeloablative SCT achieve initial CR, chronic GVHD were 29%, 4%, and 8%, respectively. With a median most relapse later on, and the long-term DFS is poor. In the past, follow-up of 9 months (2-16), the 1-year OS, EFS, NRM, relapse eff orts to intensify the conditioning regimen failed due to toxicity were 80% ± 9%, 76% ± 9%, 13% ± 7%, 10% ±7%, respectively. with signifi cant morbidity and mortality. Nowadays, most centers Causes of deaths were acute GVHD (1), relapse (1), VOD (1), CMV employ early withdrawal of immunosuppression; hence exposing pneumonia (1). the patients to risk of GVHD. Here, we report our experience in 9 Our study shows in patients with a median age of 57 years a low patients that underwent planned second allo-SCT (T2) to address incidence of early NRM (13% at 1 year) with a 1-year OS of 80%. this common scenario. These preliminary results are encouraging and a longer follow-up Patients and methods: 9 patients (4M/5F; median age 28y, 4-56) is required to accurately assess the incidences of chronic GVHD, were included. Basic disease was refractory AML (n=4), refractory late NRM and relapse. ALL (n=4) and refractory MDS\AML (n=1). Conditioning regimen

S527 for the fi rst transplant was CY-TBI in 7/9 patients. The Conditioning Conclusion: PLAS transplant using HLA matched donors is safe regimen for T2 was busulfex 3.2 mg/kg/dx2 and thiotepa 5 mg/ and assists in the control of refractory leukemia. The use of PLAS kg/dx2 in patients in CR (n=5) and melphalan 100/200 mg/m2x1 transplant with mismatched donors is associated with severe for patients with persistent disease (n=4). Transplant source was GVHD and should be used with caution. The proposed mechanism MUD (n=5), MSD (n=1), MMD (n=3). is based on TBI containing regimen in transplant 1 (myeloablative Results: the median time between the 2 transplants was 77d if possible), followed by intensifi cation of the anti leukemic eff ect (18-102d). The time for neutrophil and platelet engraftment from using another set of high-dose anti leukemic drugs diff erent from T2 was 12d (11-43d) and 38d (12-73d) respectively (fi g 1). Four the fi rst conditioning regimen, combined with allogeneic cell patients had GVHD prior to T2 (grade 1-3). There was mild-moder- therapy in the context of APCs that are changing from host type ate mucositis in all patients and self-limiting VOD in 5/9 patients. to donor type. All 9 patients developed GVHD (median grade 2) at a median of 9.5 days from the T2. 2/3 patients transplanted from MMD experienced grade 4 GVHD. There was one procedure associated toxic death. With a median of 8 months follow up, 6/9 patients are alive and 5 are disease free (fi g 2). Death causes were GVHD (n=2, both Early complications with 9/10 matching) and relapse (n=1).

R1496 Evaluation of causes of diarrhoea in peripheral stem cell transplant recipient patients L. Kaynar (1), C Pala (1), R. Buyukoglan (1), F. Kurnaz (2), G. Metan (1), S. Yazar (1), S. Sivgin (1), A. Sakioglu (1), B. Eser (1), A. Unal (1), M. Cetin (1) (1)Erciyes University (Kayseri, TR); (2)Harran University (Sanliurfa, TR)

We aimed to determine the frequency of diarrhoea occurring during the fi rst 100 days in allogeneic and autologous stem cell transplantation (SCT) patients and to reveal the microbiological causes. 452 patients who underwent transplantation due to haematological or solid organ malignancy were included. From the administration of conditioning regimen up to day 100 post-transplant, diarrhoea cases lasting at least 3 days with a minimum of 3 episodes a day were evaluated.Patient characteristics were examined: age, gender, underlying disease, type of transplant, presence of graft versus host disease (GVHD), faecal microscopy, faecal culture, faecal Clostridium diffi cile toxin, faecal cytomegalovirus (CMV) polymerase chain reaction (PCR), serum CMV PCR and fi ndings of the rectoscopic/colonoscopic biopsies.The cause of the diarrhoea could be detected in 23.3% of autologous SCT patients and in 37.2% of allogeneic SCT patients. In immunosuppressive patients who undergo stem cell transplantation, all causative factors of diarrhoea should be considered in detail, faeces analyses should be evaluated for each patient and endoscopic biopsy samples should be obtained when required. Agents that do not cause an infection in healthy individuals may lead to atypical conditions in these patients. Therefore, rarely encountered microorganisms should also be kept in mind and care should be exercised in diff erential diagnosis.

R1497 Haemophagocytic syndrome in relapsed AML patients after allogeneic stem cell transplantation C. P, M. Keklik, G. Akyol, O. Canöz, S. Sivgin, L. Kaynar, B. Eser Erciyes University (Kayseri, TR)

Hemophagocytic syndrome (HPS) induced by uncontrolled macrophage activation and subsequent graft failure is prominent complication after allogeneic stem cell transplantation (allo-SCT), a cause of severe morbidity and death, and a therapeutic challenge. A 23-year-old man without the history of familial HPS, and transplanted from unrelated fullmatch donor for acute myeloid leukemia in third complete remission, had a full engraftment at day + 15 and at day + 30 % 68 chimerism. He was relapse at day + 41. He was hospitalized and received FLAG chemotherapy and DLI. After the treatment patient had fever also on blood and permanent jugular catheter culture blastoschizomyces was observed. Patient’s permanent catheter was pulled off and amphotericin b treatment was started.

S528 Even after these treatment patients fever and pancytopenia wasn’t were in the normal range. However the potassium level still rised resolved so FLAG +28 and DLI+5 days bone marrow aspiration/ and reached to 6.5 mEq/L in the same day. Fludrocortison was ini- biopsy was performed. Macrophage predominance was observed tiated. On the 3th day of therapy potassium level was 4.9 mEq/L on bone marrow aspiration. Biopsy showed hypo-aselüler bone and decreased to 3.5 mEq/L on the 7th day and the drug was dis- marrow, increased macrophage activation and hemosiderin continued. In the following days the levels were maintained in the loaded macrophage (fi gure 1,2). Hyperferritinemia (ferritin: 4470) normal range. The patient expired due to pneumonia on day +47 was present so the patient was diagnosed with HPS and started of transplantation. dexamethasone treatment. Same day after the start of dexameth- CsA increases potassium due to mechanisms other than decreased asone treatment patients fever was resolved and on the 9.th day glomerular fi ltration. The drug inhibits Na+/K+ATPase in the of treatment neutrophil engraftment was observed. cortical collecting ducts. This leads to decreased secretion and When sustained fever with cytopenia and hyperferritinemia are contributes to clinical hyperkalemia. CsA may induce partial observed, HPS should be suspected and bone marrow aspirate tubuler insensitivity to aldosterone. In our case we could not considered. The rapid diagnosis and early initiation of an appro- determine serum minerolocorticoid level however the patient priate treatment are essential for patient management. responded well to fl udrocortisone therapy supporting secondary hypoaldosteronism induced by the drug. In conclusion although the exact mechanism of potassium elevation while using CsA remains unresolved, measures may aid in preventing this occurrence. Carefully monitoring potassium level in patients taking potassium sparing diuretics, nonsteroidal antiinfl amatory drugs, angiotensin converting enzyme inhibitors, β-blockers and in case of increased levels, witholding parenteral nutrition, giving potassium wasting diuretics and considering mineralocortico- steroids may be useful options like in our responding but fatal terminating case.

R1499 The factors related to engraftment in allogeneic transplant patients E. Uzer, I. Oner, G. Zararsiz, B. Eser, A. Unal, M. Cetin University of Erciyes (Kayseri, TR)

Introduction: We investigated the factors related with engraftment in allogeneic transplant patients. Patient and methods: A total of 100 patients who underwent AHCST in University of Erciyes, Faculty of Medicine, Haematopoi- etic Stem Cell Transplantation Unit were investigated, retrospectively. In all patient who underwent AHCST with fullmatch donors the relation between engraftment development and diagnosis, recipient age, donor age, recipient gender, donor gender, blood group compatibility, amount of CD34+ given, conditioning regimen were investigated. Results: The diagnosis of the patients were as follows: acute leukemia in 67 patients (%69,1), aplastic anemia in 7 (%7,2), non- Hodgkin lymphoma in 6(%6,2), other diseases in 17 (%17,5) (Hodg- R1498 kin lyphoma, myelofi brosis, myelodysplastic syndrome, chronic Successful treatment of cyclosporine-induced hyperkalaemia myelogenous leukemia, chronic lymphocytic leukemia, parox- with fl udrocortisone in an HSCT recipient ysmal nocturnal hemoglobinuria). 36 of patients were female T. Ozcelik, P. Seymen, F. Hindilerden, N.B. Hasbal, M. Arat (%36) and 64 patients were male (%64). 57 (%58,8) of patients Sisli Florence Nightingale Hospital (Istanbul, TR) were given non containing total body irradiation (TBI) conditioning regimen, 40(%41,2) of patients were given total body irradia- Cyclosporine (CsA) is an essential immunomodulator in HSCT with tion (TBI). The median age of 100 patients was found 33,3 years signifi cant side eff ects. It is associated with sodium retention, (±11,04). The median time of neutrophil engrafment was found hypertension, interstitial fi brosis and renal failure. Another com- 17,4±0,6 and median time of platelet engrafment was found plication is hyperkalemia. Since it is such a vital agent in trans- 12,2±0,5. The patients performed TBI neutrophil engraftment plantation discontinuing therapy is not an option. 31-year-old median value 18(15-21) between the patients performed non TBI man with acute lymphoblastic leukemia underwent allogeneic neutrophil engraftment median value 15,5(13,3-17,8) was found peripheral blood HSCT from an unrelated donor. Conditioning statistically signifi cant (p=0,005). No signifi cant relationship was regimen included TBI/Cy/ATG. CsA was initiated for GVHD prophy- found among platelet engraftment time and conditioning regi- laxis. On day +25 hyperkalemia developed. Drugs causing hyper- men (p>0,005). No signifi cant relationship was found between kalemia were ceased (spironolactone, trimetroprim). He did not neutrophil and platelet engraftment time and diagnosis, recipi- have parenteral nutrition. CsA levels were maintained between ent age, donor age, recipient gender, donor gender, blood group 200-400 ng/ml. Hyperkalemia persisted and was resistant to compatibility, amount of CD34+ given (p>0,005). loop diuretics, glucose with insulin. On day +29 potassium level Conclusion: Conditioning regimen is an important factor for reached to 6 mEq/L. At this time serum Ph, bicarbonate levels neutrophil engraftment time.

S529 Infectious diseases < 0,5 x 109/L) was 12 days. The overall incidence of fever requiring antimicrobial treatment was 34%. 12% of patients have fever unknown origin (FUO), whereas primary bacteriemia occurred in 21,3%, pneumonia in 9,7%, severe skin infection in one patient, R1500 neutropenic enterocolitis in 9,7%, acute viral B-hepatitis in 12 Clinical evaluation of secondary posaconazole prophylaxis in patients, CMV reactivation 20% in allogeneic, 0,8% (1 patient) in the initial neutropenic phase of allo-HSCT: a small case series autologous recipients. The most frequently isolated were Gram- from a single unit positive cocci from central venous line and sputum. Candida A. Tomaszewska, B. Nasilowska-Adamska, A. Szczepinski, Albicans were predominantly isolated fungi (Candida Albicans : B. Marianska Non-Albicans Candida = 54% vs 46%). Median time of febrile epi- Institute of Hematology and Transfusion Medicine (Warsaw, PL) sodes with antimicrobial therapy was 4 days (2-12 days). First-line therapy was succesful in 66% of patients. Infection-related mor- Introduction: Invasive fungal diseases (IFDs) are life-threatening tality on day +180 in our study is 10% (allogeneic:14,8% autolo- complications of allogeneic haematopoietic stem cell transplant gous: 7,5%). (alloHSCT) procedures, especially in patients with previous his- Conclusions: active disease (p=0,00078), initially neutrope- tory of invasive aspergillosis or candidiasis. There is no standard nic patients (p=0,029), graft with CD34+ cells <2,0x106/kg.tt. approach for secondary prophylaxis in this setting of patients. We (p=0,0000) in autologous and allogeneic recipients, and age up report 7 cases of posaconazole prophylaxis during neutropenic to 50 in allogeneic recipients (p=0,00016), are major risk factors phase of alloHSCT in patients with IFD history, mostly invasive in regard of infections in stem cells recipients in our transplant pulmonary aspergillosis. center. Materials and methods: We evaluated clinical results of posaconazole secondary prophylaxis in 7 patients with acute leukaemias (2-lymphoblastic, 5-myelogenous; 3 females and 4 males; R1502 median age 49; range 24–55) undergoing alloHSCT. The donors Intrathecal rituximab administration in isolated EBV-related were HLA-identical siblings (1) or unrelated (6). Peripheral blood CNS PTLD post haploidentical transplant stem cells mobilized with G-CSF were used. Conditioning regi- E. Zucchetti, G. Grillo, P. Marenco, V. Mancini, S. Jann, M. Turrini, mens were myeloablative in 6 patients and of reduced intensity I. Lotesoriere, L. Paris, P. Minga, M. Deodato, E. Morra in 1. Besides in 6 cases in vivo T-depletion with thymoglobuline Niguarda Cà Granda Hospital (Milan, IT) was performed. All patients presented with IFD during remission- induction or consolidation chemotherapy: 6-invasive aspergillo- In the setting of T-depleted Allo-transplantation standard therapy sis, 1-invasive candidiasis and aspergillosis. Patient were treated for Epstein Barr Virus (EBV) reactivation is Rituximab 375 mg/mq with various antifungals: amphotericin B, casposfungin, voricon- infused weekly until virus clearance. However, after intravenous azole and posaconazole. Two of them underwent lung lobectomy administration, this molecule reaches very low levels in cerebro- due to aspergilloma. The prophylaxis with oral posaconazole at spinal fl uid (CSF), that is therefore considered a sanctuary for the the dose of 200 mg three times daily was started in every patient therapy. directly from the conditioning regimen beginning and was con- In the case of Central Nervous System (CNS) involvement by EBV- tinued to full hematologic recovery and/or immunosuppression related Post-Transplant Lymphoproliferative Disease (PTLD), intra- discontinuation. thecal administration of Rituximab (IT-Rit) should be considered Results: 6 patients (85,7%) are alive at 7–32 months after trans- as a feasible therapeutic option. plantation and none of them developed IFD after alloHSCT. One In this report we describe a case of IT-Rit in a patient who devel- patient died on +23 day after transplant, without hematologic oped isolated EBV-related CNS PTLD after in vitro T-depleted stem recovery, due to Stenotrophomonas maltophila sepsis and fungal cell transplantation from haploidentical donor. pneumonia suspicion (not proven). No side eff ects of posacon- A 66-years-old woman underwent stem cell transplantation azole therapy was observed. from her daughter for a secondary AML in fi rst complete Conclusions: Our observational report indicates that secondary remission. prophylaxis with oral posaconazole is safe and eff ective in high On day +100 she experienced EBV reactivation with enlargement risk of mould infection reactivation group of patients during of laterocervical lymph nodes and an increasing level of blood alloHSCT procedures. These fi ndings confi rmed some previously EBV DNA up to 4300 105gEq/105 cells; we therefore administered published data, but should be studied prospectively and in the 2 doses of iv Rituximab, obtaining the adenopathy resolution and larger group of patients. the negativity of viral load. On day + 139 she developed deviation of the labial rim and gait disorders; a CT scan of the brain was performed and resulted neg- R1501 ative in terms of hemorrhages and cerebral lesions. Three days Infl uence of age, count of infused CD34-positive cells and later she was hospitalized because of worsening neurological disease activity on infections in stem cells transplanted symptoms and general conditions; subsequent tests, in particu- patients: Ten years’ centre experience lar a brain CT performed on day +151 and a magnetic resonance Z. Stojanoski, B. Georgievski, S. Genadieva-Stavrik, A. Pivkova, imaging (MRI) on day +152, revealed a mass over the sella. L. Cadievski, O. Karanfi lski, M. Ivanovski, L. Cevreska A CSF exam was performed and resulted negative for leukemic Bone marrow transplantation unit (Skopje, MK) cells, fungal and bacterial infections, but the CSF EBV PCR was strongly positive. At the same time EBV-DNA in blood was nega- Aim: We retrospectively analyzed the infective complications that tive and a CT scan of neck, thorax and abdomen didn’t reveal any occurred during a fi rst 180 days after stem cell transplantation. adenopathy. Material and methods: from September 2000 to September 2010, Considering the possibility of PTLD confi ned to the CNS, on day 163 patients (AML:82, ALL:9, CML:7, Hodgkin disease: 24, NHL:16, + 167 we administered IT-Rit, 25 mg, after standard premedica- Multiple myeloma:31, SAA: 2 CLL:1, Myelofi brosis:1), underwent tion. This procedure emerged as safe and free from immediate 173 (10 retransplant) cycles of high-dose chemotherapy and side eff ects. stem cell transplantation. 123 patients were threated with auto- Unfortunately, for worseling of general condition, patient died on logous and 50 with allogeneic stem cell transplantation. Male:93 day +171. Female:80 Median age:36 (12-63). As a source of stem cells were In this case IT administration of Rituximab was safe, either dur- used: peripherall blood in 145 (90%), bone marrow: 25 (15%), a ing the procedure or in the following days. Considering the strong combination peripherall blood/bone marrow: 3 (5%) patients. immunological fragility of these patients, when there is the sus- The patients were treated in sterile room, conditioned with picion of CNS involvement from EBV-related PTLD, we suggest to HEPA fi lters. Median duration of neutropenia ( granulocyte count perform IT Rituximab administration as soon as possible.

S530 R1503 Aug 31st 2011 from department of hematology in 36 hospitals Fatal infection with Hormographiella aspergillata in a in China. All patients received induction or consolidation chemo- leukaemic patient therapy and followed up to 6 weeks after the chemotherapy or to M. Bojic (1), B. Willinger (1), T. Rath (1), S. Tobudic (1), the next course of chemotherapy. F. Thalhammer (1), A. Böhm (2), M. Mitterbauer (1), Results: Among enrolled 1358 AML and 966 ALL patients, A. Schulenburg (1), P. Kalhs (1), W. Rabitsch (1) the incidence of IFD in AML was signifi cantly higher than that (1)Medical University of Vienna (Vienna, AT); (2)Elisabethinnen in ALL (3.3% vs. 1.4%, p=0.005), although the antifungal pro- Hospital (Linz, AT) phylaxis was more given to AML patients (29.3% vs. 23.6%). In patients received induction chemotherapy, the incidence of IFD A 61-year old female patient underwent peripheral blood stem was also higher in AML (7.4% vs. 3.8%, p=0.041), while in patients cell transplantation from an HLA-identical donor due to a relapse received consolidation chemotherapy, the incidences of IFD of acute myeloid leukaemia. On day 6 after transplantation a were similar in AML and ALL (1.4% vs. 1.3%). Multivariable logis- furuncle like lesion of about 0.5 cm diameter appeared at the tic regression demonstrated that neutropenia (absolute neutro- right forearm. The lesion expanded rapidly and became painful. phil count, ANC<500/uL) and severe neutropenia (ANC<100/uL) At fi rst fosfomycin and later also caspofungin were added empiri- were strongly correlated with the IFD (relative ratios, RR were cally. Despite this antimicrobial treatment the patient developed 7.57 and 2.20; 95% confi dence intervals were 1.85-31.01 and fever. The histopathological analysis from a biopsy revealed the 1.12-4.32 respectively) in AML patients after induction chemo- presence of fi lamentous fungi, which later was identifi ed as Hor- therapy, and use of ≥2 antibiotics longer than 7 days was risk mographiella Aspergillata. Liposomal amphotericin B was added factor for both AML and ALL patients after induction chemother- and caspofungin was replaced by voriconazol. On day +24 the 12 apy (RR=3.50 and 3.78 respectively); Moreover, the probability cm large lesion became necrotic and surgical resection was con- of IFD in neutropenic AML patients were markedly higher than ducted. One week later the patient had persistent fever, devel- that in ALL patients after induction chemotherapy (9.6% vs. 4.6%, oped atrial fi brillation and became tachypnoeic. A high-resolution p=0.033). CT scan of the lungs demonstrated multiple large nodules in both Conclusions: The incidence of IFD was also higher in Chinese lungs, surrounded by a halo of lower attenuation. Because of patients with AML than ALL, especially for patients received induc- respiratory failure non-invasive ventilation was initiated. As she tion chemotherapy. And the neutropenia in AML patients may be still had not recovered from aplasia a granulocyte concentrate the crucial reason for the higher incidence of IFD. was administered. Despite all eff orts the patient died on day +34 due to respiratory failure and septic shock. Discussion: Systemic fungal infections count among the major causes of morbidity and mortality in patients after hematopoietic Acute GvHD stem cell transplantation. Hormographiella aspergillata is an anamorph of Coprinus cinereus, which is normally found in compost and sewage. It has R1505 been rarely found in human infections and only few cases have Acute intestinal graft-versus-host disease in patients after been described in the literature. intermediate-intensity conditioning regimen (FLAMSA+RIC): Because of the rarity of cases reported there is no established diagnosis and therapy treatment for H. aspergillata. Standardized antifungal suscepti- V. Sandecka, M. Krejci, M. Doubek, Y. Brychtová, L. Cervinek, Z. Rácil, bility testing of fi lamentous fungi has been established few years M. Tošková, D. Žácková, J. Mayer ago, however there are no breakpoints for all fi lamentous fungi. University Hospital (Brno, CZ) Generally, in vitro data show low minimal inhibitory concentration for voriconazole and amphotericin B, whereas caspofungin Introduction: Allogeneic stem cell transplantation (HSCT) is an and fl uconazole reveal limited or no activity. However, outcome is intensive and potentionally curative therapy for patients with generally poor, also in patients treated with amphotericin B. Fail- hematologic malignancies. Despite prophylactic immunosuppres- ure to the therapy may be associated with delayed introduction of sive medication is acute graft-versus-host disease (aGvHD) one treatment or/and absence of recovery from neutropenia. of the main causes of therapy-related death after HSCT. Purpose: The rising incidence of rare fungal infections call for a new The purpose of this study is to prove whether clinical fi ndings approach in diagnosis and fi rst-line therapy. of aGvHD correlate with ultrasonography fi ndings and histological examination in patients with intestinal aGvHD. Patients and methods: Between May 2000 and June 2011, 105 patients R1504 received HSCT. The conditioning regimen consisted of fl udarabin Neutropenia was the most important reason for the higher AraC amsacrin (FLAMSA)- reduced intensity conditioning (RIC). incidence of invasive fungal disease in patients with acute Overall 43 of 105 (41%) patients developed aGvHD. Among them, myeloid leukaemia than acute lymphoblastic leukaemia 18 (17%) patients developed aGvHD with involvement of the Y. Ji, L. P. Xu, Y. Q. Sun, X. J. Huang intestinal tract. GvHD was diagnosed and staged according to Peking University (Beijing, CN) standard criteria. Results: We analysed 18 patients, aged 25- 62 years,in which Background and objectives: Invasive fungal disease (IFD) was developed aGvHD with involvement of the intestinal tract at a a life-threatening complication in patients with hematological median of 38 days (range: 14- 95 days) after transplantation. The malignancies after receiving intensive chemotherapy. Risk fac- clinical severity of GvHD was grade I in 1 (5.6%) patient, grade II in tors of IFD include neutropenia, use of high-dose glucocorticoid, 11 (61%) patients, and grade III in a further 6 (33.3%) patients. In use of immunosuppressants, and etc. It seems that the incidence 12 (66.6%) patients, GvHD was confi rmed histologically by biopsy of IFD in patients with acute lymphoblastic leukemia (ALL) may of the bowel. Ultrasonography examination was performed in 16 be higher than that in patients with acute myeloid leukemia (88.8%) patients.Ultrasound revealed in 9 (56%) patients bowel (AML), due to more glucocorticoid administered in induction che- irritation (thickening, dilatation and motility of the wall),with the motherapy for ALL. However, many studies showed AML patients image of pancolitis.Only 5 (23.8%) patients had GvHD confi rmed are more prone to be infected by fungi. Thus, the aim of this study by both of examination- ultrasonography and histologically by was to evaluate the incidence of IFD in Chinese patients with biopsy. The fi rst treatment in all patients was prednison at a dose acute leukemia and to explore the reasons for higher incidence of 2 mg/kg/day together with cyclosporine.Treatment was suc- in AML. cessful in 16 (88.8%) patients. For GvHD progressing on or unre- Methods: This was a prospective cohort study of consecutive sponsive to initial treatment, cyclophosfamide was administered patients with AML or ALL admitted between Jan 1st 2011 and or rabbit antithymocyte globulin.The therapeutic response was

S531 observed only in 1(5.6%) patient., other 2 (11,1%) patients with Immune reconstitution refractory intestinal GvHD died. Conclusion: As a result of this study, we suggest that ultrasonography is helpfull method for assessment of patients in whom intestinal GvHD is suspected.It should be included in the diagnostic for R1507 intestinal aGvHD, but cannot replace endoscopy and histological Graft and outcome in autologous stem cell transplantation: confi rmation of clinically suspected GvHD. a prospective GOA study E. Jantunen (1), A. Ropponen (2), T. Siitonen (3), R. Niittyvuopio (4), M. Sankelo (5), M. Putkonen (6), V. Varmavuo (1), M. Pyörälä (1), R1506 T. Kuittinen (1), R. Silvennoinen (1), A. Kutila (7), A. Nihtinen (8), Ultrasound features of severe acute graft-versus-host K. Vasala (9), P. Lehtonen (10), A. Sikiö (9), P. Valonen (11), disease involving gastrointestinal tract after allogeneic T. Nousiainen (1), J. Pelkonen (2), P. Mäntymaa (12) haematopoietic stem cell transplant (1)Kuopio University Hospital (Kuopio, FI); (2)University of Eastern A. Krasowska-Kwiecien (1), J. Gozdzik (1), S. Skoczen (1), Finland/Clinical Microbiology (Kuopio, FI); (3)Oulu University A. Dluzniewska (2), W. Czogala (2), O. Wiecha (2), Hospital (Oulu, FI); (4)Helsinki University Central Hospital (Helsinki, FI); M. Wozniak (2) (5)Tampere University Hospital (Tampere, FI); (6)Turku University (1)Jagiellonian University Collegium Medicum (Krakow, PL); Central Hospital (Turku, FI); (7)Mikkeli Central Hospital (Mikkeli, FI); (2)Jagiellonian University Childrens’ Hospital (Krakow, PL) (8)North Carelia Central Hospital (Joensuu, FI); (9)Central Finland Central Hospital (Jyväskylä, FI); (10)Savonlinna Central Hospital Graft versus host disease (GVHD) is one of the most frequent (Savonlinna, FI); (11)University of Eastern Finland/A.I.Virtanen complications after allogeneic hematopoietic stem cell transplant Institute (Kuopio, FI); (12)Laboratory of Eastern Finland (Kuopio, FI) (HSCT). When aff ecting gastro-intestinal tract, GVHD manifests as diarrhoea, abdominal pain and vomiting, which have to be distin- Introduction: Autologous stem cell transplantation is a common guished from other abdominal complications, particularly infec- practice in patients with multiple myeloma (MM) and those with tions. The aim of our study was to describe ultrasound features non-Hodgkin lymphoma (NHL).The number of CD34+ stem cells found in the bowel manifestation of advanced acute GVHD and to is considered as the main quality criteria for grafts. Blood grafts attempt to discriminate between GVHD and other bowel infl am- contain also many other cell types like various T cell subsets and mation. NK cells which may have also importance. Limited data exists in The study group consisted of 5 patients of the Department of regard to graft cellular content and hematopoietic and immune Transplantology of University Childrens’, Hospital in Cracow, recovery after high-dose chemotherapy as well as long-term out- 4 boys and one girl, aged 3 – 15 (median 8), who had under- come of ASCT recipients. gone stem cell transplant for acute lymphoblastic leucaemia Purpose: The purpose of this prospective study is to evaluate graft (2 children), aplastic anemia, chronic granulomatous disease, and content (CD34+ cell subsets, lymphocyte subsets) and hemato- Blackfan - Diamond anemia, and were aff ected by advanced acute logical and immunological recovery after transplantation and GVHD of bowel manifestation. Intestinal GVHD was assessed 3 or long-term outcome in autologous stem cell transplant recipients. 4 stage in every patient. The control group consisted of 5 healthy Patients and methods: Eligibility criteria include patients with MM children in median age of 8 years. or NHL patients who are intended for a single ASCT. After writ- In ultrasound examinations performed in patients every 3-5 days, ten informed consent, complete blood counts are evaluated on we estimated: the structure of bowel wall layers, the thickness of d +15 post graft infusion, at 1 month, at 3, 6 and 12 months post- each layer, width of bowel lumen and contents of bowel lumen. transplant. Immune recovery (CD3/CD4/CD8, CD45/CD19; CD3/ These features were examined in stomach, jejunum, ileum and CD16+CD56) will be analyzed from fresh samples by fl ow cytom- colon. Patients and controls were examined at least 12 hours after etry at 1, 3 and 6 months post-transplant. Flow cytometry will be the last meal. also performed from freezed grafts including CD34+ cell subclasses We assessed, that in GVHD patients, in every part of intestinal tract (CD34/CD38,CD117,CD133,CD45/CD19), lymphocyte subsets (CD3/ the structure of bowel wall layers become blurred. Median thick- CD4/CD8/CD45/CD19) and NK cells (CD3/CD16+CD56). Also amino- ness of ileum wall was 8 mm (versus 1,5 mm in the control group ), actinomycin D-7 staining will be performed to assess the viability of median thickness of colon wall was 10 mm (in control 2,5 mm), the cells. The patients will be routinely followed until 5 years from median dilatation of ileum was 2,9 cm (in control group 1,1 cm), ASCT in regard to late infections, relapse and death. Altogether 300 and median dilatation of colon was 6,1 cm (in control group 1,5 cm). patients will be included in fi ve university hospitals in 2012-2014; In every patient massive transsonic fl uid content was found in about 200 patients with NHL and about 100 patients with MM. bowel lumen. Also in every patient, fragmentation and desqua- Results: The study started at Kuopio University Hospital in May 2012 mation of mucosa and the presence of tissue detritus in bowel and until end of November 2012 altogether 23 patients have been lumen were described. In healthy controls no fl uid or free tissue included. The other centres have just started recruiting patients. waste in bowel lumen was present. Graft content and lymphoid recovery have been studied until now CONCLUSIONS: In our study group intestinal GVHD was deter- only in NHL patients Later patients participating in this national mined by characteristic ultrasound symtoms: the thickened, myeloma study will also be asked to participate in the GOA study. blurred wall of jejunum, ileum and colon, decay and desquama- Conclusions: The study protocol appears to be feasible and more than tion of all lenght bowel mucosa and the picture of massive exuda- 100 patients is expected to be recruited annually. Preliminary data tive enteropathy with the content of tissue detritus. We suggest on graft cellular composition, hematopoietic and immune recovery that ultrasound imaging can be the useful, immediate tool in in NHL patients mobilized with chemomobilization and those receiv- intestinal manifestation of acute GVHD diagnosis. ing plerixafor pre-emptively will be presented in the meeting.

R1508 Analysis of lymphoid subgroups and immunoglobulin levels in patients with graft-versus-host disease after allogeneic haematopoietic stem cell transplantation A. Unal, E. Uzer, Y. Koker, H. Yigit, S. Sivgin, L. Kaynar, C. Pala, M. Keklik, G. Akyol, B. Eser, M. Cetin Erciyes University (Kayseri, TR)

Introduction: We analyzed the distribution of lymphoid subgroups and immunoglobulin (Ig) levels in patients with Graft Versus Host

S532 Disease (GVHD), receiving cyclosporine treatment after allogeneic used in a stage-wise fashion, no clear guidelines for their use and hematopoietic stem cell transplantation (AHSCT). sequence are available, i.e. which one, when and what next. Patient and methods: In fourteen patients with GVHD who Objectives: The goal of this project was to provide guidelines underwent AHCST at Erciyes University Cappadocia Transplant for managing and interpreting supplementary Chm studies. The Center, Lymphoid subgroups were examined by fl ow cyto- guidelines were developed as a stage-wise algorithm presented metric analysis. Ig levels were measured by nephelometric in a decision tree, chart format to facilitate use in the laboratory. analysis. Methods: Chm results during the past 5 years, derived from three Results: Ten patients were acute myeloid leukemia (AML), four local BMT units, were reviewed regarding use of supplemen- were acute lymphoblastic leukemia (ALL), and all patients were tal studies. Cases included mixed and complete Chm, as well as receiving immunosuppressive treatment. Eight patients were malignant and non-malignant diseases. given cyclosporine treatment; six patients were given other immu- Results: Patient Chm outcomes were grouped as either: (1) Com- nosuppressive treatment. As a result of 14 patients; there were no plete Chm (0 or 100%), (2) Mixed Chm (MChm)-stable, (3) MChm- signifi cant diff erence neutrophil, lymphocyte, monocyte, eosino- unstable. We found that (a) «Single sample» Chm was only useful if phil, natural killer (NK) cells (CD56+), CD4/ CD8 ratio between the Chm = 0 or 100%, and data reliability was adequate; (b) Mixed Chm patients who were given or weren’t given cyclosporine therapy required longitudinal assessment of stability. Primary Chm results (p>0,05). In lymphocyte gate; Low HLADR expression were deter- correlated with the clinical situation in approximately 70-80% of mined in the patients who were using cyclosporine treatment and cases. In 20-30% of MChm the results raised questions requiring this diff erence was statistically signifi cant (p=0,007). There was no supplementary studies. For example, stable MChm required ML signifi cant diff erence in two group’s IgA, IgG, IgM and IgE levels and/or MRD analysis periodically to prevent “masking” of lineage (p>0, 05). specifi c changes or to interpret stable MChm in malignancy. On Conclusion: Low HLADR expressions in lymphocyte were found in this basis, we developed algorithms for the three outcome cases; patients with GVHD receiving cyclosporine treatment. the «MChm-stable» algorithm is appended. Conclusions: An algorithmic approach for selecting supplemental strategies in Chm monitoring: (1) optimizes accuracy and interpretability of results at both technical and biological levels, for almost all cases; (2) provides lab selection criteria for adjunctive Molecular monitoring tests, thus conserving resources. Despite these selection options, for routine monitoring it is important to appreciate that: (3) Data reliability assessment was helpful in all cases, but critical for single R1509 Chm values in complete Chm; (4) Longitudinal studies of MChms Quantitative chimerism monitoring: guidelines for use were critical to detect unstable trends in all cases. of supplementary studies to optimize accuracy and interpretability of results D. Kristt (1), R. Or (2), S. Israel (2), T. Lein (3) R1510 (1)Hadassah MC; Rabin MC (Jerusalem; Petach Tikva, IL); The results of HSCT in patients with EVI1-positive leukemias (2)Hadassah MC (Jerusalem, IL); (3)Rabin MC (Petach Tikvah, IL) and myelodysplastic syndromes transplanted without remission Introduction: Quantitative chimerism (Chm) monitoring, using N.N. Mamaev, A.V. Gorbunova, T.L. Gindina, E.V. Morozova, STRs, is usually based on whole blood or marrow samples. At I.M. Barkhatov, E.S. Nikolayeva, M.E. Vlasova, E.V. Kondakova, times, the results may be inaccurate, misleading or incompletely S.N. Bondarenko, N.V. Stancheva, B.V. Afanasyev refl ective of the clinical reality, so that supplemental Chm studies Pavlov State Medical University (Saint-Petersburg, RU) are necessary, such as assessing: 1) data reliability, 2) longitudinal trends, 3) MRD markers, and 4) multiple lineages (ML) [Kristt Background. Patients (pts) with higher EVI1 gene expression et al BMT, 39: 2007]. Although these ancillary analyses are often (EVI1+) may be among diff erent cytogenetic subsets of AML/MDS.

[R1509]

S533 Besides, third pts with advanced CML are EVI1-positive too. There Relapse is evidence, that HSCT in fi rst complete remission (CR) for, them is eff ective [Gröschel et al., 2011]. The achievement of CR is associated with overtreatment, which, in turn, is a problem for HSCT. Here we present the results of HSCT analysis from 18 pts with R1511 EVI1+ leukemias and MDS transplanted without CR achievement. Late relapse in patients transplanted for myelodysplastic Material and methods. In most cases the diagnosis of EVI1+ leu- syndrome: a puzzling therapeutic challenge kemias was made retrospectively. Molecular study was carried out M. Shapira (1), M. Avner (1), D. Kristt (2), S. Grisaro (1), B. Avni (1), according to Gröschel et al. AlloHSCT was done in 15 of 18 pts, L. Dari (1), I. Resnick (1), R. Or (1) whereas 2 obtained haplo- or 1 auto- HSCT. RIC or MAC condi- (1)Hadassah MC (Jerusalem, IL); (2)Hadassah MC; Rabin MC tioning regimes were chosen in accordance with pre-transplant (Jerusalem; Petach Tikvah, IL) chemotherapy character. Results and discussion. The age of pts ranged from 2 to 55 years Reccurent myelodysplastic syndrome (MDS) is not a rare condi- (mean - 28.2±3.7 y.). Among them were 6 female and 12 males tion. Relapse may occur, typically early, although late relapse (LR) with diagnoses AML, MDS, CML, and Ph+ ALL (8, 3, 6, and 1 pts, is recognized. Since a consensus approach to treatment of LR MDS respectively) with mixed karyotypes. Only 2 of them had t(9;11), as is not available, we report three LR patients, each distinguished by in recent study of Gröschel et al., whose pts were transplanted in his mode of therapy. fi rst CR. In comparison, outcome of our pts was much worse. Thus, Case 1: 41 y.o. male, diagnosed in 1996 as MDS (translocation 1;7), both our pts with t(9;11) who were treated intensively before HSCT, when he underwent non-myeloablative stem-cell transplantation died soon after haplo- and alloHSCT. Furthermore, post transplant (NST – Fludarbine, Busulfex and Thymoglobulin) from matched relapses and graft failure took place in 14 pts. It must be noticed, sibling without signs of acute or chronic GvHD. Prophylactic that post-transplant relapses in 2 pts with inv/t(3)(q21;q26) were immunosuppression for GvHD was discontinued at 3 months. successfully treated by chemotherapy, donor lymphocyte infu- Thereafter, the patient received donor lymphocyte infusion (DLI), sions and hypomethylating agents (fi g). To date 5/18 (27.8%) pts and achieved 100% donor chimerism in blood and marrow. Due are still alive. The mean OS is equal 6.0±1.2 months with range to pancytopenia, and mixed chimerism in bone marrow in 8/2011, from 0.5 to 12 months. he received a second transplant, from another matched sibling; Conclusions. EVI1+ leukemias are diagnosed among pts with dif- conditioning included Fludarbine and Busulfex. Severe mucositis, ferent cytogenetic subsets of AML/MDS and CML. HSCT in these SOS of the liver, and overwhelming sepsis developed leading to pts should be carried out in fi rst CR and supported adequately as death seven days post-transplant. before so after HSCT. An important place in this supportive ther- Case 2: 57 y.o. male with MDS (normal cytogenetics) since 1999. apy appears to be reserved for hypomethylating agents. Underwent NST from matched sibling (2003). Full engraftment occurred without GvHD. Immunosuppression was stopped one year after transplant. Pancytopenia with mixed chimerism of blood and bone marrow had appeared by 4/2011. He was treated with cycles of Vidaza combined with incremental doses of DLI. The third infusion produced acute GvHD with skin and liver involvement, which required steroid treatment. Hematologically, a persistent MDS clone was noted, with NK cell donor chimerism at 45%; the clinical status remains good. Case 3: 43 y.o male had MDS with 5q deletion. In 2005 he underwent NST from matched sibling with full engraftment without GvHD. Immunosuppression was discontinued after 4 months. Pancytopenia, mixed chimerism in blood and marrow, and recurrence of the original MDS clone, developed by 1/2008. He was treated with Revlimid and incremental doses of DLI. Chimerism became 100% donor in blood and marrow, without GvHD, and he is in excellent clinical condition on Revlimid. In conclusion, the management of patients with LR MDS is controversial. Low levels of donor NK cells, without chronic GvHD, may have prognostic as well as pathogenetic signifi cance in MDS. No single treatment emerged from this series as a recommended approach. Further study is needed for identifi cation of patients with high risk for relapse, and establishment of guidelines for prevention and treatment.

Late complications

R1512 Guillain-Barre syndrome after allogeneic haematopoietic stem cell transplantation T. Ozcelik, F. Hindilerden, NB. Hasbal, B. Topcular, A. Altinkaya, A. Kaymaz, M. Arat Sisli Florence Nightingale Hospital (Istanbul, TR)

Guillain-Barre syndrome (GBS) is uncommon during hematopoietic stem cell transplantation (HCT). The prevelance is 1% and more frequent after alloHCT. A-63-year old man with acute

S534 myeloid leukemia underwent HCT. His follow up was complicated The posttransplant complication of pulmonary aspergillosis( 20%) with gastrointestinal GVHD and was treated with corticosteroid. was signifi cant. Although matched sibling donors have tradition- On the 5th month he developed paresthesias of the feet with sub- ally been the source of donor,MUD and unrelated cord blood sequent ascending weakness and respiratory failure. He also had transplants also have good outcomes in our center. hypertension and hyponatremia. MRI of cranium and spine were normal. Cerebrospinal fl uid examination was notable for absence of leukocytes, normal glucose and increased protein concentra- R1514 tion. Nerve conduction study and electromyelography were Elevated serum ferritin as a risk factor of organ toxicitiy consistent with GBS. He was treated with intravenous immuno- in children treated with haematopoietic stem cell globulin. After therapy he became normotensive, hyponatremia transplantation: preliminary results resolved and neurologic symptoms improved. He was discharged A. Zaucha-Prazmo (1), K. Drabko (1), J. Gozdzik (2), M. Wozniak (2), however 2 weeks later he was admitted with neurological dete- J.R. Kowalczyk (3) rioration and pneumosepsis. He required mechanical ventilation. (1)Medical University (Lublin, PL); (2)Medical University (Kraków, PL); Antimicrobial therapies, inotropic agents and plasma exchange (3)Medical University (Lublin, PL) were initiated. His neurological condition improved again and was extubated. However the fi fth plasma exchange session was Iron overload is a relatively common complication in adult patients complicated with anaphylactoid reaction with subsequent entu- undergoing haematopoietic stem cell transplantation (HSCT). Ele- bation and mechanical ventilation. He expired due to sepsis. No vated pre-transplant ferritin levels may increase the risk of post- infectious etiologic agent was identifi ed for the cause of GBS. transplant organ toxicity in this group of patients. Both EBV-DNA and CMV-DNA were negative. Serum autoanti- The aim of the study was to assess the frequency and signifi cance bodies to ganglioside antigens were negative. In the transplant of elevated ferritin serum levels before transplant procedure and setting reported cases of GBS were related with infectious agents in early post-transplant period in children treated with HSCT such as CMV, EBV, HHV-6. The reported incidence of GVHD among because of malignant and non-malignant diseases. patients who develop GBS is 73%. Donor derived alloreactive B Patients: A total number of 31 children (21 boys and 10 girls) with cells may cause GBS as the cases respond to immunsupressive median age of 10 years (range 1-17), treated with HSCT were therapy. The prognosis is particularly poor after HCT. In our case included into the study. In 19/31 patients HSCT was performed GVHD was under control and no infectious agent was identifi ed. because of malignant disease (ALL- 7 pts, AML– 2 pts, NBL- 4 pts, This supports the fact that there is a greater clinical heterogeneity HD- 2 pts, Sa Ewing– 2 pts, NHL- 1 pt, JMML- 1 pt; in 12 pts because in these patients and the pathogenesis is still less certain. of non-malignant disease (FA- 3 pts, CGD- 3 pts, SAA- 2 pts, WAS- 1pt, inborn errors- 3 pts). Methods: Serum ferritine levels were estimated before conditioning and 3 weeks post-transplant. Organ toxicities observed in postransplant period were graded according to WHO Regimen Paediatric issues Related Scale. Results: Median concentration of ferritin was signifi cantly higher before transplantation in patients with malignant as compared to R1513 non-malignant diseases (1037 mg/dl vs 103 mg/dl; p=0,03). Three Haematopoietic stem cell transplants for patients with bone weeks post transplantation this diff erence was no longer signifi - marrow failure: results from a single paediatric centre cant (2520 vs 1482; p=0,28). Organ toxicities (renal, hepatic) more A.M. Tan, K. Vijaya, M. Chan than grade I WHO occurred in twelve children. Ferritin concentra- KK Women’s and Children’s Hospital (Singapore, SG) tion in patients , who developed toxicities was higher as compared to those without such problems, however the diff erence was not Severe aplastic anemia(SAA) is common in Asia Pacifi c area. Based signifi cant (2608 vs 1531 mg/dl; p=0,053). on epidemiology study SAA is 2 to 3 times higher in Asia than the Patients with high pre-transplant ferritin concentration (more West. Hematopoietic stem cell transplantation(HSCT) is the treat- than 1000 ng/ml) experienced toxicities in 54%, while in children ment of choice for bone marrow failure conditions in children. We with ferritin less than 1000 ng/ml this complication occurred in describe the results of HSCT for childhood bone marrow failure 31% (ns). from a single paediatric transplant center. Conclusions: Our preliminary results suggest, that children with Method: Allogeneic HSCT for 11 patients with bone marrow fail- malignant diseases are at risk of iron overload before transplanta- ures were performed over a period of 9 years from March 2003 to tion, and this may contribute to post-transplant organ toxicities. October 2012.There were 6 males and 5 females. The underlying Further study are necessary to determine particular risks of hyper- conditions were SAA 7(64%),hypoplastic anaemia evolving into ferrytinemia in children undergoing stem cell transplantation. SAA 1(9%), Fanconi’s anaemia(FA) 2(18%) and Paroxysmal nocturnal hemoglobinuria(PNH) 1(9%).8(73%) patients underwent HSCT from matched related donors.3(27%) patients were from unrelated cord blood/marrow donor (UCB/MUD). Conditioning regimes for 10 (90%) patients consisted of cyclophosphamide (CPA)and ATG ± Thoraco-abdominal Irradiation (T.A.I). 1 pt with PNH had busulphan in addition. Recently we have changed to fl udarabine based conditioning regime in place of TAI. Graft vs host disease (GVHD) prophylaxis consisted of cyclosporine ± methotrexate, ± methylprenisolone(for UCBT), ± mycophenolate moetil. Results: All engrafted well and to date remained disease–free. The median neutrophil engraftment was D+14 (range D+12 to D+17). The median platelet engraftment was D+20 (range D+16 to D+35). The median follow up duration was 939 days(range from 84 to 3521). There was no graft failure and no transplant related mortality (TRM). Post HSCT complications were: AGVHD grade 1-2(skin)2, Chr GVHD GIT(gut)1, autoimmune haemolytic anemia/thrombocytopenia 1, BK virus infection 1, CMV infection 3, CONS infection 2, Strept viridian 1 and pulmonary aspergillosis 2. Conclusion: HSCT with MSD or unrelated cord/donor for bone marrow failure conditions yielded excellent results in our center.

S535 R1515 and 5 umbilical cord blood. 4 children was transplanted in fi rst Micafungin for antifungal prophylaxis in paediatric complete remission (CR1) (2 patients with slow response to the allogeneic haematopoietic stem-cell transplant recipients initial course of inducion and 2 patients with BCR-ABL gene rear- J. Horakova, S. Sufl iarska, I. Bodova, A. Chocholova, P. Svec, rangement), 22 children was in second complete remission (CR2), Z. Laluhova-Striezencova, O. Fabri 4 in third remission (CR3) and 5 received a HSCT with positive Comenius University Children’s Hospital (Bratislava, SK) minimal residual disease (pMRD). Results: With a median of follow-up of 108.6 months (range 5-234) Introduction: Fungal infections are associated with high morbid- 14 patients were alive and free of leukemia and 21 died (19 died ity and mortality in patients after hematopoietic stem-cell trans- of relapse-related causes and 2 from graft- versus-host disease plantation (HSCT). High risk of IFI after HSCT is a consequence of (GvHD). Acute GvHD was developed in 58.1% (50% of matched many factors. Among pathogens arises the proportion of Can- related donor and 73% of matched unrelated donor) with skin dida nonalbicans strains, invasive aspergillus, and rare fungi. The involvement in 94.4% and gut or liver involvement in 55.6%. The necessity for new antifungal prophylaxis emerges for paediatric acute GvHD was grade I/II in 61.1% and grade III in 38.9%. Chronic HSCT patients. GvHD was developed in 29% of the patient, and the aff ected Methods: Authors present experiences with micafungin prophy- organs were: skin 77.7%, liver 33.3%, lung 33.3%. Global grading of laxis in pediatric HSCT recipients at Bone Marrow Transplantation chronic GvHD: mild 37.5%, moderate 25% and severe 37.5%. None Unit in Bratislava, Slovakia. In a period of 44 months (January 2009 relapse was found in HSCT-CR1 group and 53.8% relapse in HSCT ≥ – August 2012) 56 patients, 38 boys and 18 girls, aged 3 months CR2 group. All pMRD children prior HSCT relapsed. The estimated – 18 years, received HSCT for malignant and non-malignant dis- overall survival (OS) and disease-free survival (DFS) were 41 and eases. 38/56 patients received graft from unrelated donor – 23 39% with a “plateau” extending from 2.75 to 20 years. patients MUD 10/10, 10 patients MUD 9/10, 5 patients MUD 8/10. Conclusions: The majority of the paediatric patients who receive a 18 patients underwent HSCT from family donor – 17 patients from HSCT do it in CR2. The patients with positive MRD before the HSCT MSD, 1 from HLA-identical mother. Micafungin was started at evolve unfavorably. The HSCT in CR1 was indicated in few cases D -1; the weight adjusted standard dose was given once daily as (very high-risk ALL and slow response to the remission induction a 1 hour infusion. Prophylaxis continued from day -1 to engraft- phase of therapy) with good results. ment and restitution of oral intake. Median of engraftment was 19 days (11-34 days); duration of prophylaxis was 15-52 days. The tolerance and side eff ects were monitored by clinical examination, laboratory fi ndings (whole blood count, biochemical analysis, microbial cultivations, and mycotic antigens evaluation), and Accreditation and regulation other examinations based on a patient condition. The micafungin prophylaxis was satisfactory in 37/59 patients (66%). In 3/56 patients micafungin dose was adjusted from 1mg to 2 mg/kg/day R1517 due to prolonged febrile neutropenia without further need of The use of electronic medication order entry system for antifungal drug change. In 8/56 patients micafungin was stopped paediatric conditioning regimens in Spanish hospitals – in 6/56 due to possible IFI, 1/56 for probable IFI, and in 1/56 S. Valero García, P. Marrero Álvarez, P. Escobar Cava, F. Fuentes patients for proven IFI – candidemia non albicans. In 7/56 mica- Socorro, J. M. Fernández Navarro fungin administration was interrupted due to veno-occlusive dis- H. U y P La Fe (Valencia, ES) ease of the liver (VOD), the multifactorial HSCT complication. With the exclusion of 7 VOD patients from analysis, micafungin was Introduction and objectives: Electronic medication order entry well tolerated in all 49/56 patients and antifungal prophylaxis was systems with clinical decision support (EMO/CDS) have been eff ective in 37/49 patients (75.5%). There were no deaths associ- designed to reduce both, prescription mistakes and adverse ated neither with VOD nor IFI in our cohort. events. Moreover, it contributes to standardize prescriptioncon- Conclusion: The increasing resistance and changes in spectrum of sidering it is build buildon the basis of consensual protocols. Our causative pathogens has to be taken into account while choos- hospital has adopted this system for pediatric stem cell transplant ing treatment strategy. Echinocandine micafungin showed to be a (SCT)conditioning prescription since 2011. suitable and effi cient antifungal prophylaxis in HSCT children. The aim of this study is to update the situation and compliance of Spanish pediatric hospitals toEMO/CDS in the pharmacotherapeutic process (prescription, pharmaceutical validation, prepara- R1516 tion and administration) and we also give an overview of our own Review of acute lymphoblastic leukaemia in children under- activitywith EMO/CDS. going haematopoietic stem cell transplantation: 20 years of Methods: We designed a simplequestionnaire regarding EMO/ experience in our centre CDS implementation: A.I. Rodríguez, A.I. Pascual, A. Contento, D. Bardán, J. Coín, 1) Does your hospital have an EMO/CDS for SCT?If affi rmative M.E. González, A.I. Heiniger answer: does it have clinical decision support system? Hospital Materno-Infantil Carlos Haya (Málaga, ES) 2) Does it capture analytic data? 3) Is there any transcription during the pharmacotherapeutic pro- Background: Acute lymphoblastic leukemia (ALL) is the most com- cess? If affi rmative answer: which one? Which professional does it? mon malignancy of childhood. With current therapy, adapted to We identifi ed 24 Spanish hospitals with pediatric stem cell the risk, the complete remission is reached in 95 % of the cases and transplant activity in the 2011 Spanish National Transplantation a prolonged remission in 60-70%. A small percentage of paediat- registry and pharmacists from our Unit phoned thepharmacy ric patients with ALL are going to need hematopoietic stem cell departmentsof these 24 selected hospitals. The obtained informa- transplantation (HSCT) to achieve fi rst or successive remission. tion was compiled and analyzed. Patients and methods: A descriptive and retrospective study of 35 Results: Twenty-one hospitals answered our questionnaire. Only paediatric patients (24 male, 11 female) diagnosed with ALL who four out of 21 interviewed hospitals (19%)implement EMO/CDS, underwent HSCT, in the last 20 years in our centre. The median but just one of them capture analytic data. There is not any tran- age was 6.4 years (range 0,5-14). By immunophenotyping the scription during the pharmacotherapeutic process, except for one 86% of ALL was developed from B- cell precursors (Common-ALL: hospital.The rest of institutions (17) still use a paper ordering sys- 61,9%) and the remained of T-cell origin. Only 3 patients have tem and a pharmacist carries out the transcription process.Three central nervous system involvement at diagnostic. The modalities of themuse hand written paper orders along the pharmacothera- of HSCT were: 4 Autologous-SCT and 31 allogeneic-SCT: 15 from peutic process. The rest workwith diff erent electronic devices to matched unrelated donor and 16 from matched related donor. perform pharmaceutical validation and elaboration. Transcription The source of stem cells was: 21 bone marrow, 9 peripheral blood of elaborating process is necessary in 4 hospitals.

S536 In our hospital, EMO/CDS system (Farmis R) hasbeen implemented Acute leukaemias with 19 computerized conditioning protocols linked to 14 diagnoses. So far, we have prescribed, validated and elaborated the conditioning protocols of 49 patients. Conclusion: Despite the fact that EMO/CDS improves patient R1519 safety and standardizes clinical practice, it is not yet a common Comparative outcome of reduced intensity and myeloabla- resource for pediatric SCT in Spanish hospitals. In our experience, tive conditioning regimen in HLA identical sibling allogeneic once the system is learned, the pharmacotherapeutic process is haematopoietic stem cell transplantation for acute leukae- faster, safer and it helps professionals to feel more confi dent. mia patients: a single centre experience H. Goker (1), E. Ozdemir (1), B. Uz (1), Y. Buyukasik (1), M. Turgut (2), S. Serefhanoglu (1), S. Aksu (1), N. Sayinalp (1), I. C. Haznedaroglu R1518 (1), F. Tekin (1), S. Unal (1), O. Bektas (1), E. Eliacik (1), A. Isik (1), Chronicles of a brand new private stem cell transplantation O. I. Ozcebe (1) unit in Turkey: nine months’ experience (EBMT CIC 862) (1)Hacettepe University (Ankara, TR); (2)Ondokuz Mayis University MK Yüksel (1), M Kurdal (1), Ö Köktas (1), D. Özbek (1), I. Tek (1), (Samsun, TR) G. Kilic (1), O. Ilhan (2), Ö Arslan (2), M Özcan (2) (1)Private Medicana International Ankara Hospital (Ankara, TR); Objectives: Due to the high transplant related morbidity and mor- (2)Ankara University School of Medicine (Ankara, TR) tality (TRM), relatively younger acute leukemia patients that have a good performance status and no comorbidity are eligible for The number of stem cell transplantations in Turkey in 2000 was myeloablative conditioning (MAC) followed by allogeneic hema- approximately 300.In 2011, partly due to the fi nancial support for topoietic stem cell transplantation (allo-HSCT). This retrospective transplantations provided by the Ministry of Health and Social comparative study assessed the outcomes of 84 Turkish adult Security Administration, it went up to 2000. The populaition of acute leukemia patients who received either RIC or MAC condi- Turkey is more than seventy million and this transplantation tioning regimens for allo-HSCT. count is still not enough. Methods: The outcomes of 84 consecutive adult patients with Stem Cell Transplantation Units are rapidly growing in Turkey, ALL (n=38) or AML (n=46) who underwent allo-HSCT from their there were only 38 in 2004; now, in 2012, there are 57 adult HLA-identical siblings were evaluated. The median age at trans- and pediatric transplantation centers .They can be categorized plantation was 34 years (17-58) for the whole patient population. into three: Of these, 24 patients received a MAC and 60 patients received a 1-The State Hospital Bone Marrow Transplantation Units n=5 fl udarabine-based reduced intensity conditioning regimen (RIC). (8,7%) Results: The median age at transplantation was signifi cantly high 2-The State University Hospitals n= 36 (63,2 %) in the RIC patients compared to the MAC patients (36 (17-58) vs 27 3-The Private University and Hospitals n=16 (28,1%) years (17-47), p=0.002). Eleven of the RIC patients (18.3%) and 6 Aim: To present the transplantion activity of a new established of the MAC patients (25%) developed acute graft-versus-host dis- adult transplantation center. ease (GvHD). Seventeen of the RIC patients (33.3%) and 4 of the Results:At Private Medicana International Ankara Hospital Bone MAC patients (16.7%) developed chronic GvHD. Overall, the TRM Marrow Transplantation Unit we have performed seventy trans- at day 100 was 1.7%; the TRM increased to 5.1% at 1 year. After plantations in nine months, 60 of which are autologous, 9 allo- a median follow-up of 32 months (range, 1-119), for the entire geneic full match and 1 haploidentical transplantations.The fi rst group, the 3-year estimated overall survival (OS) was 57,5% and transplantation was an autologous on the twenty fourth of Febru- the disease-free survival (DFS) was 51,5%. The OS for ALL and AML ary 2012. At the end of six months, in September 2012 we started patients were 53.9% vs 62.1%: and DFS were 50.5% and 53.4%, allogeneic transplantations, and recentley in November we per- respectively. The 3-year estimated OS for RIC and MAC patients fomed a haploidentical transplantation. 6 of the seventy patients were 63.2% and 41.7%; and DFS were 57.1% and 34.7%, respec- died resulting in –transplantation related mortality of 8.5 %. The tively. In ALL patients, conditioning regimens (RIC vs MAC) led to causes of death were progressive disease in two of the cases bac- similar OS and DFS; however, in AML patients both OS (70.1% vs terial sepsis in 4 cases. The transplantations performed divided 21.4%) and DFS (59.3% vs 42.9%) were found to be higher in RIC into types of disease along with gender distribution are given in patients compared to MAC recipients. In multivariate analysis, the table below. the diagnosis (p=0,03) and RIC regimen (p=0,027) were the prog- Conclusion: Although Private Medicana International Ankara nostic variables for prolonged OS in all patients; and RIC regimen Hospital Bone Marrow Transplantation Unit was established very (p=0,031) was the only prognostic factor for prolonged OS in AML recently (in January 2012), it is now in the top 10 centers in Tur- patients. CR1 was correlated with a prolonged DFS as an indepen- key and is expanding very fast. Our aim is to be one of the larg- dent variable for all patients (p=0,09). est transplantation centers not only in Turkey but also in Asia and Conclusion: This retrospective study, demonstrates that RIC con- Europe. ditioning regimens may provide a longer OS and DFS, especially in patients with AML who are in fi rst CR, not eligible for MAC conditioning.

R1520 Role of allografting in acute lymphoblastic leukaemia: 12 years’ experience L. Giaccone (1), B. Bruno (1), E. Maffi ni (1), E. Audisio (1), S. D’Ardia (1), D. Caracciolo (1), B. Allione (1), A. Busca (1), F. Ferrando (1), F. Testa (1), M. Festuccia (1), L. Brunello (1), M. Boccadoro (1), U. Vitolo (1), R. Passera (2), M. Falda (1), F. Marmont (1) (1)Divisions of Hematologies, Città della Salute e della Scienza di Torino (Turin, IT); (2)University of Turin (Turin, IT)

Acute lymphoblastic leukemia (ALL) is a rare disease in adults and clinical outcomes remain unsatisfactory. Aim of the study: To evaluate the experience on 84 consecutive patients diagnosed with ALL at the Divisions of Hematology, S. Giovanni Battista Hospital, Torino, between 1999-2011.

S537 Methods: Patients, median age 49 years (18-73), were treated R1522 according to Center guidelines or on clinical trials active at the Allogeneic haematopoietic stem cell transplantation for time of diagnosis. High risk patients younger than 60 were con- Philadelphia or BCR-ABL-positive acute lymphoblastic sidered for allografting (SCT) as part of fi rst-line treatment. Treat- leukaemia: experience of the Centre National de Greff e ments for 80% (20/25) of patients with traslocation (9;22) also de Moelle, Tunis, Tunisia included tyrosine kinase inhibitors after 2002. R. El Fatmi, B. Ladeb, S. Menif, N. Ben Abdeljelil, L. Torjemane, Results: Overall, 25% (21/84) presented with leukocytosis and 56% A. Lakhal, T. Ben Othman (38/68, not done in 16) had poor-prognosis cytogenetic abnor- Centre National de Greff e de Moelle Osseuse de Tunis (Tunis, TN) malities. Complete remission (CR) was achieved in 92% of patients (77/84). After a median follow-up of 5 years (3-143 months), 43% Objectives: The prognosis of Philadelphia-Positive Acute Lympho- of patients are alive, with a median overall survival (OS) and event- blastic Leukemia (ALL Ph1) is poor. We describe the outcome of free survival (EFS) of 2 and 1,6 years, respectively. Forty patients Allogeneic Hematopoietic Stem Cell Transplantation (AHSCT) for received an allograft in fi rst (no.=32) or second (no.=7) complete patients with ALL Ph1 and we evaluate minimal residual disease remission, or in progression (no.=1), from a HLA identical sibling (MRD) before and after AHSCT. (no.=21), an unrelated (no.=17) or a related other than sibling Methods: Our retrospective study enrolled all consecutive patients (no.=2) donor. Cumulative incidences of acute and chronic graft- aff ected by de novo ALL Ph1 and or bcr-abl+ who underwent versus-host disease were 47% and 39%, respectively. Median OS genoidentical AHSCT between November 2003 and June 2012. All and EFS in patients who received SCT were 60 months and not the patients except one received imatinib in the pre-transplanta- reached, respectively, whereas in those younger than 60 who did tion treatment. Conditioning regimen consisted of fractionated not undergo a SCT median OS and EFS were 16 and 13 months TBI and Vepeside. Graft versus host disease (GVHD) prophylaxis (p=0.024 and 0.026) respectively. Stratifying patients by year of associated cyclosporine ± short course methotrexate. MRD was diagnosis, a trend toward improved EFS in those diagnosed after assessed in blood by qualitative or quantitative PCR. 2008 as compared to those diagnosed before 2003 (2.3 years Results: Eighteen patients were transplanted during the period of versus 1.3) was observed, whereas OS remained super-impos- the study (12/18 patients between 2010 and 2012). Median age at able. By multivariate analysis, the prognostic role of leukocyto- transplantation was 36 years (range: 6- 47). Fifteen (83%) patients sis at diagnosis was confi rmed for both OS and EFS (HR 2.48, IC were transplanted in fi rst complete remission (CR), whereas 3 95% 1.28-4.80, p=0.007, and HR 3.72, IC 95% 1.93-7.14, p<0.001, were transplanted in advanced phases (2 in 2nd CR and1 in blastic respectively). Furthermore, in younger patients, an advantage phase). Eight patients (44%) had positive MRD at the time of trans- in those who received an allograft in terms of both OS (HR 0.11 plantation. After a median follow-up of 9 months (range: 2-39), CI 95% 0.03-0.44 p=0.002) and EFS (HR 0.12 CI 95% 0.03-0.47 6 patients (33%) died from toxic cause and one patient developed p=0.002) was seen. isolated central nervous system relapse. Twelve patients are alive Conclusions: SCT played a signifi cant role in ALL patient survival. (66%): 10 with negative and 2 with positive MRD. Overall Survival Future trials that combine allografting with tyrosin kinase inhibi- (OS) and Disease Free Survival (DFS) were respectively 56% and tors or novel monoclonal antibodies, such as blinatumomab, and 50% (83% and 72% for patients transplanted after 2010). Fourteen evaluate treatment response by molecular methods may further patients were evaluable for quantitative MRD: bcr-abl transcript improve clinical outcomes. persisted undetectable in all patients transplanted with negative pre-transplant MRD, whereas among the 7 patients transplanted with positive MRD, the bcr-abl persisted detectable in 1 patient R1521 and became undetectable in 6 patients after a median follow up Impact of bone marrow transplantation on outcomes in of 4 months. Reappearance of bcr-abl transcript was observed in 3 patients with acute lymphoblastic leukaemia: a comparison cases (two were in complete molecular remission after imatinib) between cytoreductive and conventional chemotherapy Conclusion: In our experience, the recruitment and the prognosis A. Ghavamzadeh, K. Alimoghaddam, S. Soleymanzadeh, S. Mousavi, of ALL Ph1 / bcr-abl + was improved since 2010. B. Bahar, A. Jalali, M. Jalili, L. Sharifi Aliabadi, R. Maheriazar, A. Karimi, E. Eini, M. Jahani Tehran University of Medical Sciences (Tehran, IR) R1523 Eff ective post-remission therapy with an autologous stem Aim: Allogeneic hematopoietic stem cell transplantation (allo- cell transplant (ASCT) for the ELN-favourable and intermedi- HSCT) is a potent cure for acute lymphoblastic leukemia (ALL) and ate-risk acute myeloid leukaemia (AML) patients use of reduced intensity conditioning regimens can reduce HSCT- J. Cerny, M. Ramanathan, G. Raff el, A. Evens, W. Walsh, N. Fortier, related drug toxicity. The study assesses the role of this modality. L. Shanahan, J. Bednarik, A. Rosmarin, R. Nath Methods: Newly diagnosed ALL cases who candidate HSCT with University of Massachusetts (Worcester, US) full HLA matched sibling after obtain informed consent received reduced induction chemotherapy (RIC) (Vincristine/Dexametha- Introduction: The 5-year disease free survival (DFS) typically sone) and underwent early HSCT after disease stabilization within ranges 60% - 80% while the overall survival (OS) typically ranges 15-30 days without intention to achieve CR. These cases compared from 20% - 50%, without advantage of allogeneic transplantation with patients who received conventional chemotherapy. for the favourable and intermediate-I ELN risk AML groups (Doh- Results: Twenty eight patients whose transplanted after RIC ner et al Blood 2010; 115: 453-474). compared with 70 patients treated conventionally. The mean Methods: We have retrospectively analyzed the outcome of 18 age was 24.7 year (SD=7.25) for the transplant group and 28 consecutive patients (pts) with favorable or intermediate-I ELN year (SD=7.22) for the chemotherapy group. Relapse after HSCT risk AML who received high dose cytarabine (HiDAC) and anthra- occurred in 3 (15%) of transplant and 46 (65.7%) of chemotherapy cycline based induction chemotherapy and underwent high dose group. Two year months leukemia-free survival was 50.1% (SE: chemothrapy with autologous stem cell transplantation (ASCT). 11.8%) in transplant and 42.6% (SE: 6.6%) in the chemotherapy All patients with CN-AML were off ered to receive azacytidine and two year overall survival was 50.5% (SE: 13.2%) and 52.8% maintenance after ASCT, one patient opted out. (SE: 6.5%) in transplant and chemotherapy groups, respectively. Results: Median age was 60 years (range, 29-80); 8 were (44%) Death occurred in 8(35.7%) of transplant (causes: relapse, GvHD) female. Eight (44%) had CN-NPMmut, 8 had CBF-AML (44%) and and 42(60%) of chemotherapy group (causes: relapse). 2 (11%) patients had CN-AML. All patients had received HiDAC Conclusion: Reduced induction followed by early transplantation based induction followed by at least one cycle of HiDAC based without consolidation is not clinically inferior then chemotherapy consolidation. All achieved complete remission (CR) and pro- alone however relapse rate and death was higher in chemother- ceeded with stem cell mobilization. Mobilizing chemotherapy was apy group and is clinically signifi cant. A randomized study with HIDAC (1-3 grams/m2 for 6-8 doses)/Etoposide (15- 40 mg/kg) in suitable case size is recommended. 17 (94%) patients and G-CSF alone in one (6%) patient. Median

S538 time from diagnosis to ASCT was 4.2 (range 3.6-7) months. As con- transplantation (alloHSCT). However time of performing alloHSCT ditioning 13 (72%) pts received Busulfan (3.2 mg/kg x 4)/Etopo- and effi cacy of the usage diff erent of conditioning regimen is side (60 mg/kg) and 5 (28%) received high dose melphalan. The controversial. median CD34 cells infused was 4.8 x 106/kg (range 2.8 to 15.9). The aim: To estimate results of alloHSCT in patients (pts) wih AML All patients engrafted, with median days to neutrophil recovery exclusive of M3 FAB variant. of 11 (10-13). Median time to platelet (20 x 103/mL) engraftment Methods: From January, 2002 to December, 2011 alloHSCT were was 21 (15-40). The median length of inpatient stay post trans- performed in 65 pts (33 girls and 32 boys) with median age 10,5 plant was 14 days (10-22). One patient died of progressive disease years (range 1-20 y.o.). 20 pts were in 1 complete remission (CR), 14 months post ASCT. Two patients died in remission, one on day 19 pts were in 2 CR, 26 pts were in progression disease or 3 CR (sal- 53 (sepsis) and the other on day 836 (unknown cause) post ASCT. vage groupe). 11 pts had secondary AML. 48 pts were after unre- With the median follow-up of 21 (range 2- 43) months post ASCT lated allo HSCT, 17 pts- after related. Bone marrow (BM) used in the 3-year disease free survival (DFS) is 92% and the 3-year overall 31 pts, peripheral blood stem cells (PBSC) in 34 pts. Mieloablative survival (OS) is 72%. conditioning regimen (MAC) applied in 31 pts, reduced intensity Conslusions: In our experience, combination of HiDAC based conditioning (RIC) in 34 pts. MAC consisted Busulfan 16 mg/kg induction with ASCT and maintenance azacytidine provides safe + Cyclophosphamide 120 mg/m2. RIC included Fludarabine 150 and eff ective disease control in pts with favorable or intermedi- mg/m2 + Melphalan 140 mg/m2 in 15 pts, Fludarabine 150 mg/ ate-I ELN risk AML. m2 + Busulfan 8mg/kg in 18 pts, FLAG in 1 pt. Acute graft versus host disease (aGvHD) prophylaxis was by Cyclosporine (CsA) and short course of MTX in 38 pts, CsA +MMF in 6 pts, Tacrolimus and R1524 MMF–9 pts. 2 pts after related allo HSCT and all pts after unrelated Mutations in the kinase domain of FLT3 confer resistance to HSCT recieved ATG 60 mg/kg. Median CD34± 5,9*10/6kg. the FLT3 inhibitor and associate with poor outcomes follow- Results: Engraftment was at day + 18 (range 10-30). Primary non ing conventional chemotherapy engraftment was 11 % (7 pts, 4 pts from this group had second- J. Kwon, Y. Koh, J. Kim, K. Ahn, D. Lee, S. Yoon ary AML). 8- years overall survival (OS) in all group was 45 %. In 1 Seoul National University College of Medicine (Seoul, KR) CR OS was 68%, in 2 CR – 52%, in salvage group -22%. OS in pts after MAC was 58%, after RIC 32%. We did not reveal signifi cant Objectives: FMS-like tyrosine kinase 3 (FLT3) has emerged as a diff erence in OS between children and adolescents: 48% vs 39%; promising molecular target of treatment. Recent study suggested between BM and PBSC: 48% vs 39%; between related or unrelated that FLT3 could be alternatively activated by mutations in the donor: 50% vs 42%. OS in pts with secondary AML was 22%. Main kinase domain such as D835Y. It suggested that FLT3 mutations causes of death were relapse (13 pts), aGvHD (11 pts). Transplant could limit the eff ect of FLT3 inhibitor to the subset of leukemic related mortality (TRM) and relapse risk in salvage group ver- cells which harbor FLT3 mutations. The study was designed to sus 1 and 2 CR was 40% vs 26% (Er=0.06), 32% vs 15% (p=0,03), examine the FLT3 mutations of leukemic cells at initial diagnosis respectively. and at relapse in comparison with corresponding germ line con- Conclusion: allo HSCT is eff ective in children and adolescents with trol from saliva samples. AML, the best results in OS observed in pts with 1 and 2 CR. Pts Methods: We utilized FLT3 gene target sequencing to distinguish with high risk AML need to search suitable donor (related or unre- FLT3 mutations that contribute to FLT3 inhibitor (AC220) activ- lated) and performe allo HSCT in 1 CR for improvement results ity. To examine the eff ects of FLT3 mutations on FLT3 inhibitor- and decreasing mortality. mediated apoptosis, cell proliferation assay was performed using primary cultured leukemic cells from AML patients. Results: We identifi ed 12 cases of FLT3 mutation-related relapse in R1526 AML patients [52 patients, 17 set of constitutive samples: at diag- Granulocytic sarcoma: usefulness of ultrasound for detection nosis, complete remission, and relapse]. Unique seven FLT3 muta- and analysis of microvascular changes with a second genera- tions at initial diagnosis and four (N676, F691L, L676K, D835Y) at tion contrast media enhanced ultrasound sonography and relapse were founded. Four diff erent somatic mutations of FLT3 comparison with extramedullary plasmocytoma: two distinct gene infl uenced overall survival. Inhibition of cell proliferation by enhancement patterns AC220 occurred in approximately 50% of primary cultured leu- E. Benedetti, F. Caracciolo, E. Orciuolo, G. Buda, M. Petrini kemic cells. AML cells with FLT3 mutation including N676, F691L, Hematology Unit (Pisa, IT) G697R, L676K, and D835Y were resistant to AC220 treatment. Four FLT3 mutations might be associated with poor outcomes follow- Introduction: Granulocytic sarcoma (GS) represents an extramed- ing cytarabine and anthracyclin-based induction chemotherapy. ullary proliferation of myeloid blasts, common in the pediatric age We further screened FLT3 mutations which were associated with group (incidence up to 30%) more than in adults (2–5%). GS may relapse, in 2 independent adult normal karyotype AML patient present concurrently with a new diagnosis of AML or as evidence cohorts, of 217 at initial diagnosis and 87 at relapse. of disease recurrence, as de novo disease, or at relapse after an Conclusion: Our study will contribute to elucidate the role of FLT3 allogeneic transplant (Tx). Outcomes are usually poor. Extramed- mutation on relapse, progression and response to treatment in ullary plasmocitoma (EP) is a plasma cell tumor, without any signs AML patients. of systemic spread. Both may present with masses involving all body sites. Color Doppler Ultrasound (CDU) detects arterioles while Contrast Enhanced Ultrasound Sonography (CEUS) detects R1525 real-time capillary microvascular (MV) blood fl ow and overall Allogeneic haematopoietic stem cell transplantation in chil- microvascular network (MN). Methods. In the last 7 years 7/120 dren and adolescents with acute myeloid leukaemia. Single pts with GS were evaluated (5.8% incidence). Pts were studied centre experience with standard US and CEUS to assess: i) the usefulness of US to N. Stancheva, E. Semenova, A. Borovkova, O. Paina, S. Razumova, detect GS, ii) the MV enhancement pattern (EN) with CEUS of GS T. Bykova, P. Kozhokar, A. Rats, K. Ekushov, I. Markova, A. Shetsov, (MN), iii) we also compared the EN of GS with 11 cases of EP to S. Bondarenko, L. Zubarovskaya, B. Afanasyev test the hypothesis that MV changes in GS and EP might reveal R.Gorbacheva Memorial Institute of children hematology and diff erent EN pattern refl ecting diff erent neoangiogenesis activity. transplantation (St.Petersburg , RU) Results: Sites involved in GS: stomach (n=1), abdominal lympho- nodes (n=3); soft tissue masses (n=2), para-vertebral mass (n=2). Background: The treatment of children and adolescents with In 2/7 pts GS occurred as relapse after allogenic BMT (preceding acute myeloid leukemia (AML) has improved considerably over bone marrow relapse by 1.5 and 3 weeks respectively); in 3/7 GS the past decades. It is connected with introduction more intensive was found at diagnosis, in 2/7 was diagnosed at relapse. US fi nd- chemotherapy and using allogeniec haematopoietic stem cell ings: GS and EP appeared as hypo-echoich, solid masses with

S539 regular shaped border (superimposable). CDU showed modest in most cases. The probability of being a donor-derived constitu- diff erences between GS and EP (few vs. absence of arterioles in Gs tional abnormality is fairly rare; but extremely important to recog- vs. EP). CEUS: in GS absence of vessels within the masses, at excep- nize in donors and recipients. tion of few capillary and sometimes arterioles fl owing through the Case report: a 61-year-old man was referred for management of mass with per-lesional EN. This pattern was consistent during fol- refractory AML. He had been diagnosed with AML (FAB-M2), by low up until either progression or remission. On the contrary CEUS morphologic and immunophenotypic bone marrow (BM) analy- in EP showed intense EN with a rich MN covering the whole lesion. sis. He had received 2 cycles of FLAG (fl udarabine, cytarabine, Conclusions: US in AML might not only be considered as routine G-CSF) but failed to achieve complete remission. His disease examination but also a useful tool focused to detect GS especially failed to respond to various lines of salvage therapy namely in the setting of refractory/relapsed AML pts and in high risk pts CLAG (cladribine, cytarabine, G-CSF), 5-Azacitidine, and MEC post allogenic Tx. CEUS showed a peculiar enhancement pattern (mitoxantrone, etoposide, cytarabine). He received HLA-matched in all GS consistent with many avascular areas fi lled with leukemic (10/10,A,B,C,DRB1,DQ), G-CSF mobilized, peripheral blood stem cells, without a strong neoangiogenesis as opposed to multiple cells from his brother following a preparative regimen of fl uda- myeloma which is confi rmed to have an intense neoangiogenesis rabine (30mg/m2/day x 5 days), intravenous busulfan (130mg/ capillary network. m2/day x 4 days) and 4Gy TBI (in 2 doses) on day -1; and GVHD prophylaxis consisting of cyclosporine and methotrexate. His pre- transplant chromosomal analysis was consistent with male karyo- R1528 type. Neutrophil and platelets engraftment occurred on days +15 Diff erences in Bcl-2, Bax, CD95, ACE expression on CD34+ and +10 post allografting, respectively. Day +30 work up showed cells of bone marrow and peripheral blood of healthy donors normal CBC and BM analysis consistent with complete hemato- and acute leukaemia (AL) patients logic remission. Conventional cytogenetic analysis identifi ed a E. Parovichnikova, E. Khodunova, I. Galtzeva, L. Mendeleeva, new abnormality t(1;8)(p36;q24) in 19 metaphases. BM CEBPA gene S. Kulikov, V. Savchenko mutation analysis was negative. In light of previously normal cyto- National Research Center for Hematology (Moscow, RU) genetic analysis as well as evidence of post-transplant remission, there was a high level of suspicion that t(1;8)(p36;q24) might be Diff erences in Bcl-2, Bax, CD95, ACE expression on CD34+ cells of of donor origin. Accordingly, conventional chromosome banding bone marrow and peripheral blood of healthy donors and acute of donor peripheral blood cells performed on day +68 confi rmed leukemia (AL) patients. In healthy donors the % of CD34+ cells presence of same abnormality. Donor chimerism measured on expressing Bcl-2, Bax, CD95 is much higher in PB, than in BM. unsorted BM, on Day +98 post allografting, showed 99.3% donor So we can speculate that PB precursors are more defended from cells. The patient was started on maintenance 5-azacitidine since diff erent infl uences in blood stream. day +60. He remains without evidence of disease, now over 100 In contrast to donors the % of CD34+ cells in PB and BM express- days post-transplantation. ing investigated Ags is similar in AL patients. So we may suppose Conclusion: We aim at raising awareness of the presence of con- that there is no bone marrow/ vascular barrier in acute leukemia stitutional, non-pathogenic, chromosomal aberrations that may and CD34+blast cells entering the circulation are equivalent to transfer from donors to recipients. Performing chromosomal anal- residing in b/m. ysis on donors, as in our case, helps to avoid misinterpreting the The % of ACE expressing CD34+ cells in BM of AL pts is very similar presence of such mutations as evidence of cytogenetic progres- to healthy donors while in PB of healthy donors it is much higher sion of disease. suggesting the recirculation of hematopoietic precursors respon- sible for long-term reconstitution. So, our data demonstrate that Bcl-2, Bax, CD95 and ACE expres- R1530 sion on CD34+ cells in AL pts and donors signifi cantly diff ers. It’s 5-azacitidine is a possible therapeutic option for patients also worth to note that donor PB and BM CD34+ cells are not simi- with relapsed AML or MDS after allogeneic stem cell lar regarding investigated markers indicating that circulating and transplantation residing in BM CD34+ cells has diff erent expression profi le. N.K. Steckel, T. Gromke, Y. Hegerfeldt, M. Hlinka, M. Koldehoff , M. Ditschkowski, A. Elmaagacli, D.W. Beelen Universityhospital Essen (Essen, DE)

Objectives: Patients with relapsed AML or MDS after allogeneic stem cell transplantation (aSCT) have a very poor prognosis, attaining rarely a long lasting complete remission. Usually reduction of immunosuppression in combination with donor lymphocyte infusions (DLI) is used to induce an antileukemic eff ect. Intensive chemotherapy may be an option in patients with high blast counts. A second aSCT can be performed with curative intent only in small cohort. All options are associated with a high treatment related mortality and a low response rate. 5-azacitidine (5-Aza) is a DNA hypomethylating agent which induces remissions in AML and MDS. Because of its R1529 low toxicity it is a safe alternative for older and strongly pretreated Development of translocation (1;8) following allogeneic patients. We wanted to know if 5-Aza can be a feasible and eff ective haematopoietic cell transplantation for acute myeloid therapy for patients with relapsed AML and MDS after aSCT. leukaemia: a constitutional abnormality of donor origin Methods: 10 patients (6 female, 4 male) with relapse at a median Z. Otrock (1), S. Fares (2), R. Mahfouz (2), A. Bazarbachi (2), time of 261 days after transplant (range 44-1210 days) were M. Kharfan-Dabaja (3) treated with 5-Aza. Eight patients were transplanted for AML, (1)Cleveland Clinic Taussig Cancer Institute (Cleveland, US); and 2 patients for MDS at a median age of 58 years (range 44-68 (2)American University of Beirut Medical Center (Beirut, LB); years). Two patients were transplanted with an identical sibling (3)H. Lee Moffi tt Cancer Center and University of South Florida donor, one with a haploidentical family donor, and 7 patients with College of Medicine (Tampa, US) matched unrelated donors. All patients had chemotherapy based conditioning regimens. Background: detection of cytogenetic abnormalities is not Results: 5-Aza was administered in all patients for a median of unusual in relapsed acute myelogenous leukemia (AML) following 5 courses (range 1-6). All patients received additional therapy allogeneic hematopoietic cell transplantation (allo-HCT). These with DLI, and one patient with FLT3-positive AML Sorafenib, abnormalities represent evolving or expanding leukemic clones, respectively. Five patients had a good response with decrease of

S540 blast count in the peripheral blood after at least one course. In all summarizes the result of post-transplant treatment with MSCs of a these patients donor chimerism increased with 3 patients devel- 26-year-old aplastic anemia patient complicated by invasive sino- oping a complete, and 2 patients 95% donor chimerism. All these orbital aspergillosis. The patient treated with MSCs to expedite the patients are in ongoing complete haematologic remission. Two dual eff ects of MSCs in immune reconstitution: suppression against patients developed chronic GvHD of the skin or the mucosa. An alloreactive T cells and facilitation of re-engraftment process. This additional patient just commencing therapy also shows declining complicated patient did not developed acute and chronic graft blasts the peripheral blood and an increase in donor chimerism. versus host disease. Aspergillus infection healed completly. The Four patients had progressive disease under 5-Aza and died in engraftment failure also recovered without any complication. consequence of persisting relapse. During his current visit at fourth year after transplantation, he was Conclusion: 5-Aza in combination with DLI or targeted drugs can in hematological remission. Human MSCs seems to have a role on be a feasible and eff ective therapy for patients with relapsed AML prevention, or overcoming immunological complications in the and MDS after aSCT. patients who undergo stem cell transplantation.

R1531 Allogeneic stem cell transplantation for blastic plasmacytoid dendritic cell neoplasm Autoimmune disorders T. Heinicke (1), H. Hütten (2), T. Fischer (1) (1)University Hospital (Magdeburg, DE); (2)Klinikum Braunschweig (Braunschweig, DE) R1533 Non-myeloablative allogeneic stem cell transplant in Introduction: Blastic plasmacytoid dendritic cell neoplasm an idiopathic thrombocytopenic purpura patient: (BPDCN) is an often rapidly fatal hematologic disorder charac- a case report terised by skin infi ltrates with or without bone marrow or lymph C.P. Pala (1), R. Buyukoglan (1), Y. Altuner (1), F. Kurnaz (2), node involvement. Most patients reported in the literature were L. Kaynar (1), S. Sivgin (1), M. Keklik (1), B. Eser (1), M. Cetin (1) treated with either acute lymphoblastic leukemia (ALL)-type, (1)Erciyes University (Kayseri, TR); (2)Harran University CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) (Sanliurfa, TR) or CHOP-like combination therapies. The median overall survival of BPDCN patients with conventional chemotherapy is only 12 to Patients with refractory chronic autoimmune thrombocytopenia 14 months. However, patients treated with allogeneic stem cell (ITP) have a signifi cant risk of morbidity and mortality related to transplantation (alloSCT) seem to have a better overall survival. hemorrhage (1). ITP in adults is usually chronic, idiopathic and fre- Case Report: Patient 1 was a 21-year-old female that presented quently refractory to conventional treatments (2). Allogeneic hae- in the 33rd week of her fi rst pregnancy with lower back pain for matopeietic stem cell transplantation (HSCT) is feasible for patients which no cause could be diagnosed. A Caesarean section was with refractory chronic ITP, a substantial proportion of whom may performed. However, the back pain persisted. Shortly thereafter obtain durable remissions (3). We report a case of a man with ITP, severe pancytopenia developed and bone marrow biopsy as well refractory to the conventional therapy, submitted to HSCT, which as fl ow cytometry showed BPDCN with a 95% infi ltration. There obtained long term stable response on platelet count. was no skin involvement. The treatment consisted of three cycles of chemotherapy according to the German ALL Study Group protocol 07/2003 leading to fi rst complete remission (CR). The patient was then transplanted from an unrelated donor after myeloablative conditioning. With a follow up of 15 months after transplant Genetic disorders the patient has remained in CR. Patient 2 was a 63-year-old female presenting with multiple skin lesions at her left lower leg that were diagnosed as being BPDCN without bone marrow involve- R1534 ment. First CR was achieved with 6 cycles of CHOP chemother- Clinical impact of infused T-cell numbers after MUD stem cell apy. Consolidation radiotherapy (40 Gy) was applied thereafter. transplants in oncohaematological children However, only four months later distant relapse at multiple skin V. Grassi, C. D’Ippolito, L. Palumbo, R. Ceresoli, S. Cavagnini, sites was diagnosed. Three cycles of salvage therapy were applied V. Bennato, E. Soncini, F. Ricci, L.D. Notarangelo, M.F. Schumacher, which led to second CR. The patient was then transplanted from R. Baff elli, F. Bolda, M. Cossandi, M. Zucchi, L. Rugaba, A. Beghin, an unrelated donor after reduced intensity conditioning (RIC) A. Lanfranchi, F. Porta consisting of fl udarabine and thiotepa. The patient has remained Children’s Hospital (Brescia, IT) in complete remission for more than 9 months after transplant. Conclusion: The cases presented here suggest that alloSCT after either We have analyzed 19 onchaeamtological children who under- myeloablative or RIC conditioning is an active form of consolidation went MUD BMT who presented a GVHD grade III/IV children try- in BPDCN in fi rst or second CR. Prospective studies are warranted in ing to fi nd an impact of HLA mismatching, source of cells and age order to clarify the role of alloSCT in the treatment of BPDCN. A GVHD grade III/IV in our experience do not exceed 10% of the case since the vast majority of the cases has less f 3 yrs of age. We monitored the absolute number of T cells infused. We graft children weighting less than 10 KG with BM stem cells, PBSC in all other cases. In 2011, 22 children received transplant, 7 received Aplastic anaemia PBSC with a mean number of CD34+ cells of 25.6x106/Kg and CD3+ cells 42.1 x 106/kg, while 10 children received BM stem cells received a mean number of CD34 + cells 8,2X106/Kg and R1532 T cells 38.7 x 106/Kg remaining within a number of Tcells ranging Use of MSCs to off er dual benefi ts of immune suppression or between 30 and 70 x 10 6/Kg. Nevertheless 4 out of 22 (18%) pre- immune reconstruction in a patient with aplastic anaemia sented A GVHD >grade III, 2 despite a 6/6 matched donor, 2 with complicated by invasive sino-orbital aspergillosis an antigenic mismatch A or B, 2 out 4 died. All the 4 patients were H. Ozdogu (1), M. Yeral (1), C. Boga (1), I. Ilknur Kozanoglu (2) grafted in presence of an infectious agent (bacterial and/or viral). (1)Hematology (Adana, TR); (2)Physiology (Adana, TR) We retrospetively revised children (40) who presented A GVHD > Grade III below the age 3 yrs old grafted in our Center and realized Cultured human bone marrow mesenchymal cells (MSCs) have that the mean number of CD3+ cells weren’t that diff erent from immunomodulatory and tissue regenerative properties. This report the 2011 group: 48,9 x 106/Kg. The HLA disparity moreover wasn’t

S541 diff erent between the two groups.Moreover the stem cell source with homogenous parenchymal echo and spleen was greater than doesn’t seem to have an impact as well. In conclusion in our expe- normal. On the peripheral blood smear, a few atypical monocytes rience expecially in primary immunodefi ciencies (PID) aff ected were detected; thus, bone marrow aspiration and biopsy were per- children undergoing BMT, the most relevant criterion to foresee formed. On bone marrow smear, elements from three cell lineages an increase risk of GVHD is the infectious state at time of trans- and sea-blue histiocyte infi ltration were detected.Result of biopsy plant and the onset of viral or bacterial infections after transplant. was reported as Nieman-Pick disease. Sphingomyelinase activity Our policy, nevertheless, is not to administrate more than 70 x 10 (7.73±3.08 nmol/17 hours/mg protein) in leukocyte was found as 6/Kg T cells and do not accept a donor who shows more than one 1.38 nmol/17 hours/mg protein. Thus, patient was diagnosed as HLA antigenic mismatch. Type B Niemann-Pick. Allogeneic stem cell transplantation was considered for Niemann-Pick disease; however, this consideration was discarded because of the problems which might occur due to R1535 immunosupression during transplantation period and incompli- Bone marrow involvement of a patient with Niemann-Pick ance, given the mental status and bronchiestasis in the patient. disease concomitant with Kartagener’s syndrome: report In conclusion Kartagener’s syndrome should be considered as a of a rare case possible diagnosis in patients with bronchiectasis and it should M. Keklik be kept in mind that multiple syndromes may exist together in Erciyes University (Kayseri, TR) cases presented with recurrent respiratory tract infection in the presence of splenomegaly. Niemann-Pick disease is a rare lipid storage disorder with autosomal recessive inheritance, which is characterized by accumulation of sphingomyelin and other sphingolipids in macrophages. Kart- agener’s syndrome is a rare syndrome with autosomal recessive inheritance consisting of chronic paranasal sinusitis, situs inversus and bronchiectasis. It is accounted from half of primary ciliary dys- kinesia syndromes. Here, we reported a case having Kartagener’s syndrome with concomitant Niemann-Pick disease, as there is no report of such case in literature. A 21-years old woman was referred to hematology department due to fi nding of splenomegaly from another facility where she presented with cough, abdominal pain and fatigue. It was also found out that dextrocardia was detected on the chest radiography which was performed 4 years ago when she presented with recurrent sinusitis and cough. We also learned that, on the thorax computerized tomography (CT) performed after confi rmation of dextrocardia by echocardiography, there was inverse positioning of liver and spleen and appearance of bronchial dilatation, peri- bronchial thickening and consolidation favoring bronchiectasis at lungs. She was diagnosed as Kartagener’s syndrome due to presence of bronchiectasis, situs inversus and sinusitis. On abdominal ultrasound evaluation, it was found that liver was normal in size

Lymphoma

R1536 Treatment of high-risk T-NHL with autologous or allogeneic stem cell transplantation C. Busemann, S. Klein, CA Schmidt, M. Evert, G. Dölken, W.H. Krüger Ernst-Moritz-Arndt-University Greifswald (Greifswald, DE)

Purpose: The prognosis of peripheral T-cell lymphomas (PTCL) and other advanced stage T-cell lymphomas is poor. Here we present the results of high-dose therapy and autologous (autoSCT) or allogeneic stem cell transplantation (alloSCT) obtained in 18 patients with T-cell neoplasia. Patients and methods: 18 patients with a median age of 47,6 (range 20,5-64,1) years were treated with autoSCT (n=6) or alloSCT (n=12) from 1996 to 2011. All patients were at high risk either due to the IPI-score or to the fact that SCT was part of a salvage therapy. Conditioning prior to alloSCT was myeloablative

S542 in six cases (50%). The patients were heavily pre-treated with 9 Prognostic Index (IPI) was considered to be the most important (median, range 2-38) cycles of chemotherapy. prognostic factor for survival and the strongest indicator for iden- Results: Nine patients are alive in complete remission after a tifi cation of high-risk patients, who are unlikely to be cured with median follow-up of 11,3 months (range 0,8-137,7). The estimated standard chemotherapy. Having in mind that IPI is based on 5 overall survival was 51% after one year and 38,3% after ten years. clinical characteristics (age,performance status, stage, extranodal There was a trend for better survival after related alloSCT (80% involvement, LDH level) and it is constructed in the pre-rituximab at 10 years) compared to the other modalities. Manifestation of is clear that R-IPI should be tested in rituximab era to provide any GvHD did not infl uence survival. information of its validity. Conclusion: AlloSCT from related donors can cure patients from We retrospectively analyzed unselected population of 80 patients PTCL or advanced other T-cell lymphomas. Unrelated alloSCT or with confi rmed diagnose of diff use large B cell lymphoma treated high-dose therapy and autoSCT are an option for patients without at University hematology department in the period of 2005- a familiar donor. 2010. All patients were uniformly treated with R-CHOP regiment as initial treatment with curative intent. There were 80 patients with mean age 54, 5 years (15-84), male 35 and female 45. Older R1538 than 60 years were 29 patients (36, 25%). More than half of the High-dose therapy and autologous haematopoietic cell patients (42) were diagnosed in advanced stage of the disease. We transplantation in angioimmunoblastic T-cell lymphoma analyzed fi ve prognostic factors: age, performance status, stage, M. M. Herráez Albendea, R. De Paz Arias, M. Morado Arias, F. J. Alves extranodal involvement, LDH level and through the multifactorial Ferreira, M. A. Canales Albendea analyses we selected two groups of patients. One with 0 to 2 fac- La Paz, Hospital (Madrid, ES) tors as patients with low risk. Patients with more than 3 factors are considered as high risk. There is statistically signifi cant diff erence Objective: Angioimmunoblastic T-cell lymphoma (AITL) is a rare in overall survival between two groups with fi ve –years overall subtype of peripheral T-cell lymphoma that carries a poor prog- survival 70% for low risk patients and 47% for high risk. High-risk nosis. The aim of this study was to evaluate the eff ect of high-dose patients may be candidates for autologous transplantation as therapy (HDT) followed by autologous stem-cell transplantation initial treatment, having in mind that in the rituximab era relapses (ASCT) on patients with (AITL) occur very early in the fi rst year and are diffi cult to be treated. Methods: We analyze retrospectively 12patients (5 male , and 7 R-IPI score is signifi cant predictor and should be used for risk strat- female) treated in our center for a period of eight years (2002-2010) ifi cation of patients with aggressive B-cell lymphoma. However, with a mean age of 75 years (range 40-84). Of these patients (p), these fi ndings should be validated prospectively in an indepen- 1 patient (8.33%) presented Ann-Arbor staging I-II and 11 p dent population of patients. (91.66%) staging III-IV. 8 patients (66.66%) presented systemic symptoms (“B”) and 10p (83.33%) were IPI>3. 5p (41.66%) were treated with CHOP like regimens, 1patient (8.33%), was treated R1540 with Cyclophosphamide-Cladribine-Prednisone, and all of them A novel conditoning regimen for primary CNS lymphoma: with ASCT. high dose rituximab, cyclophosphamide, etoposide and Results: After a median follow-up of 90 months (range, 19-120), BCNU (RCVB) 5p ( 41.66%) remained alive, and 7p (58.33%) died. 4p (57.14%) MK Yüksel (1), I. Tek (1), Ö. Köktas (1), M. Kurdal (1), SK. Toprak (2), patients died as a result of disease progression, and 3p (42.85%) O. Ilhan (3), Ö. Arslan (3), M. Özcan (3) died as a result of regimen-related toxicity. The overall survival (1)Private Medicana International Ankara Hospital (Ankara, TR); (OS) was 6p (50 %) at 24 months and 5p (41.66%) at 48 months. (2)Baskent University School of Medicine (Ankara, TR); (3)Ankara A 41.66 % of patients achieved complete response, 8.33 % partial University School of Medicine (Ankara, TR) response. Patients who received a transplant during fi rst complete remis- We present three cases whom were diagnosed primary Cen- sion had signifi cantly superior progression-free survival (PFS) and tral Nervous System (CNS) lymphoma, who recieved high dose overall survival. The PFS for patients who received their trans- chemotherapy followed by autologous transplantation. Case1: plants in complete remission were 66.66 % and 66.66 % at 24 and A 45 year old man admitted to hospital because of memory loss. 48 months, respectively; 50% and 0% for patients who received Kranial MR showed a mass in the frontal lobe. The histopathologic their transplants in partial remission. examination of excisonal biopsy was diff use large B cell lymphoma Conclusion: Limitations of this study include the number of (DLBCL).After two cycles of fi rst line therapy(*), the progenitor cells patients included. This study shows that HDT and ASCT off ers the were mobilized (**) and 12.5 x 10e6/kg CD34+ hematopoetic pro- possibility of long-term disease-free survival to patients with AITL. genitor cells were collected. The response was more than partial Early transplantation is necessary to achieve optimal results. response after two cycles.After the fourth chemotherapy as soon More eff ective and novel therapies are needed for patients with as the recovery, the patient underwent autologous stem cell trans- high-risk disease. plantation. The conditioning regimen was high dose RCVB (***) Case2 A 58 years old woman admitted to the hospital bacause of left hemiplegia.The huge mass in the temparoparietal lob of R1539 the brain was DLBCL. After two cycles of fi rst line chemotherapy R-IPI in selecting patients with diff use large B-cell 4.24 x 10e6/kg CD34 + hematopoetic stem cells were collected. lymphoma Complete remission was achieved after four cycles and the patient S. Genadieva Stavrikj, Z. Stojanoski, A. Pivkova Veljanovska, underwent autologous trnasplantation as soon as possible. Case3 S. Krstevska Balkanov, B. Georgievski A54 years old woman admitted our hospital bacause of refractory University Hematology Clinic (Skopje, MK) DLBCL of CNS. She had recieved one cycle of chemotherapy in another center.She recieved one cycle of rituximab containing reg- Nowadays, goal of treatment approach in diff use large B cell lym- imen and 20% increased dose methoteraxate and ARA-C.Followed phoma is cure and fi rst step towards it is to achieve complete by this CD34+ hematopoetic stem cells 6,87 x 10e 6/kg were col- remission. DLBCL is a potentially curable disease, with curability lected. She had still progressive disease after this cycle. As soon as highly dependent on clinical and biological features. According to the peripheral blood recovered, she underwent autologous trans- the WHO classifi cation of Hematological Malignancies, the entity plantation.There was more than %90 reduction in the mass, after of DLBCL is characterized by rapidly growing mature B cell tumors transplantation whole brain radiotherapy was started. with large or relatively large cells and encompassing several dis- Results: The most common side eff ect was emesis. There was tinct clinopathologic diseases, several diff erent histologic variants no grade 3-4 non hematologic toxicity. Neutrophil and platelet and clinical subtypes.Diff erent subgroup of patients with DLBCL engraftment was on day 10,9; 10,13; 9,10 for the cases 1, 2 and needs diff erent treatment. In the pre-rutuximab era International 3 respectively.

S543 infused with a CD34+ cells total dose of 6x106. The engraftment was reported on 14th day. No occurence of acute graft versus host disease as well as no incidence of clinically extensive chronic disease were reported. Further bone marrow examinations confi rmed complete haematological and cytogenetic remission after Allo HSCT with a full complete donor chimerism until November 2010, when a decrease chimerism donor cells below 60% was determined in absence of signs of disease progression. Subsequently, blood cell count showed again macrocytic anemia and bone marrow biopsy documented erythroid dysplasia and a new del(5q) clone arose fi nally at FISH analysis. A need of at least one unit of RBC for month became evident in Juanary 2011 when the chimerism analysis showed the presence of 50% donor cells. The patient started Lenalidomide at the dose of 10 mg/day for 21 days. She achieved transfusion independence after 8 weeks of treatment. No severe myelosuppression occurred in the course of treatment and other adverse events were low or moderate. After 4 months, the patient achieved an increase donor chimerism to over 90% and FISH analysis performed after 3 months of Lenalidomide treatment, showed a complete remission. To date complete clinical response, transfusion independence and cytogenetic remission are still reported and further analysis showed a continuous increase of donor cells chimerism to over 95%. Lenalidomide is the treatment of choice for lower-risk anemic, transfusion-dependence patients with del(5q) karyotype who do not respond to an ESA or who have high endogenous serum erythropoietin levels. Our case showed that Lenalidomide is also a viable strategy for cure relapse after an Allo HSCT with an acceptable safety profi le. Larger prospective studies are required for confi rming the role of Lenalido- mide given alone or associated with the lymphocytes infusion in this setting of patients.

R1542 A case of allogeneic stem cell transplantation in a patient diagnosed with primary myelofi brosis and history of portal Conclusion: It is a well tolerated, targeted, feasible therapy and hypertension secondary to splanchnic thrombosis highly eff ective even in refractory CNS lymphoma. M. Zapata, K. Galvez, F. Combariza *First line chemotherapy was 4 cycles of Rituximab 375mg/m2/ d Hospital Pablo Tobon Uribe (Medellin, CO) -5 and 0, methoteraxate 3.5 g/m2 (d1),ARAC 2x2g /m2 (d 2,3) **Mobilisation protocol : Peripheral blood progenitor stem cells We report about a woman of 53 years old for fi rst time met in were collected from the patients’ after the second cycle when the our center by the hepatology group 3 years ago. The patient was bone marrow was recovering from chemotherapy. referred to our center with the diagnostic of portal and mesen- ***Conditioning regimen was high dose RCVB: Rituximab teric bed thrombosis occurred 7 years ago, from since she was 375 mg/m2 (-7), BCNU 500 mg/m2 (-6), Etoposide 200 mg/m2 anticoagulated with vitamin K antagonist. The patient had no his- (-6,-5, -4), Cyclophosphamide (-6,-5,-4,-3) tory gastrointestinal bleeding. The hepatology group in the fi rst evaluation found a patient without stigmata of cirrhosis but with a grade II splenomegaly, they tried to evaluate with abdominal Doppler ultrasound that shows recanalization of the splenic. An upper endoscopy that showed esophageal varices. The study Myelodysplasia for intrinsic hipercoagulable state was discarded. But the initial cell blood count with mild leukocytosis and persistent platelet count above 500,000 that were not expected in a patient with the R1541 degree of portal hypertension and the splenomegaly. The bone Lenalidomide as salvage therapy in a case of relapse after marrow study reports a chronic myeloproliferative neoplasm, allogeneic HSCT in a patient with myelodysplatic syndrome primary myelofi brosis (mild to moderate fi brosis) with normal with isolated 5q deletion (5q-) karyotype and JAK2 mutation negative. Meanwhile, she was A.M. Carella (1), E. Merla (1), M.M. Greco (1), G. Sanpaolo (1), treated with prednisone and thalidomide for 7 months limited by L. Savino (1), G. Perla (1), G. De Cillis (1), M. Di Candia (2), a peripheral neuropathy secondary to thalidomide and remained N. Cascavilla (1) in prednisone treatment. During this time the patient presented (1)IRCCS “Casa Sollievo Della Soff erenza” (San Giovanni Rotondo, recurrent upper gastrointestinal bleeding secondary to esopha- IT); (2)Medical Aff air Celgene srl. (Italy, IT) geal varices that require band ligation. Two years ago the patient received bone marrow stem cells (2,63x106/kg CD34+) from a We reported a case of a patient with 5q- syndrome who achieved 10/10 matched HLA identical related donor. The non mieloabla- stable complete cytogenetic remission and increase donor chime- tive conditioning regimen consisting of Busulfan, Fludarabine rism with lenalidomide as salvage therapy after a relapse post Allo and Thymoglobulin. The graft-versus-host disease (GVHD) pro- HSCT performed during the pre-lenalidomide era. In February phylaxis included in a methotrexate and cyclosporine regimen. 1994, a 41-years old female had a diagnosis of 5q- syndrome. Tak- Engraftment with absolute neutrophil counts > 500/ μ l occurred ing into consideration the progressive anemia that increasingly on day +12. required transfusion support and after an observation period, In the post-transplant the patient has a persistent pancytopenia the patient was scheduled to undergo an Allo HSCT in June 2002. with a transfusion requirement weekly. In the +81 day the chime- Cy plus TBI were used as conditioning regimen and PBSC were rism with a 100% but the pancytopenia persists. One year ago, in

S544 a staff decision between hepatology and hematology services, it R1544 was considered prudent to exclude the spleen with endovascular Mobilization of autologous peripheral blood haematopoietic therapy for persisting splenomegaly and repeated bone marrow stem cells for newly-diagnosed multiple myeloma treated study. The endovascular therapy was able to reduce splenomegaly with cyclophosphamide, thalidomide and dexamethasone and ascites was resolved. The bone marrow study reports a nor- regimen, experience in Macedonia mal bone marrow without fi brosis. The patient was asymptomatic B. Georgievski (1), L. Cevreska (1), Z. Stojanoski (1), S. Genadieva without ascites and a total independence of transfusion support. Stavrik (1), L. Chadievski (1), R. Grubovic (2), S. Useini (2), The response to allogeneic stem cell transplantation was good in A. Pivkova Veljanovska (1) a patient with severe Portal hypertension (1)University Hematology Hospital (Skopje, MK); (2)Institute for Transfusion Medicine (Skopje, MK)

Background: A variety of factors have been reported to aff ect prognosis in patients with multiple myeloma. CTD regimen has been Myeloma known as an eff ective induction therapy in patients with newly diagnosed MM. But the results for successful stem cell mobilization after this initial treatment is still inconsistent. The aim was to R1543 identify the infl uence of this regimen on stem cell yield. Initial experience of using G-CSF only for autologous stem Material and methods: A total of 55 patients with MM were treated cell mobilization in patients with multiple myeloma with autologous transplantation at our hospital. 25 MM (45.4%) M.P Putkonen, K.R Remes, M.K Kauppila, U.S Salmenniemi, patients received CTD regimen as induction treatment in a dura- M.I-R Itälä-Remes tion of 6 cycles. Stem cells were mobilized with G-CSF as single Turku University Hospital (Turku, FI) agent mobilizing regimen. Results: The median age at diagnosis was 52 years (range 37- Background: Cyclophosphamide (CY) + granulocyte growth factor 65). Median duration from start of CTD to fi rst collection was 13 (G-CSF) has been the standard mobilisation regimen for collection months (range 5-17). At mobilization the response rate of CTD of autologous stem cells in multiple myeloma (MM). G-CSF alone was 10% CR, 25% VGPR and 63% PR. Median number of harvested has the benefi t of more accurate timing of stem cell aphereses. CD34+cells/kg was 3.2x10(6)/kg at fi rst attempt. 82% of patients In Turku, autologous stem cell transplantation (ASCT) for MM has reached the minimal PBPC target of 2.0x10(6)/kg CC34+ cells and been performed since 1992. With the standard mobilisation a fail- 55% of patients achieved collection of >4x10(6)/kg with median ure incidence in newly diagnosed patients has been 7 %. In year of 3 apheresis procedure (range 1-6).At the end of second mobi- 2011 we moved to use G-CSF only for mobilisation. lization 90% of patients had yield of >2.0h10(6)/kgCD34+ cells. Aim of the study: To test the feasibility of use of G-CSF only for During mobilization three patients developed grade ¾ non- mobilisation of stem cells in patients with newly diagnosed MM hematologic adverse events. with special reference to the additional need of plerixafor. Conclusion: CTD regimen is an eff ective induction therapy for Patients and methods: The fi rst ten consecutive patients mobilised newly diagnosed MM patients showing high response rate with G-CSF at a dose of 10 μ g/kg/d are analysed. The median age acceptable for autologous stem cell yield without any impairment was 61 (range: 44-69) years and time from diagnosis to mobilisa- following stem cell collection. tion 4 (3-8) months. The preceding anti-MM therapy was VEL-DEX for 9 patients and REV-DEX for one patient. G-CSF was begun on Friday, and blood CD34+ cell measurements R1545 were begun on Monday (Day +4). Aphereses were initiated when Multiple myeloma: study of diff erent transfusional blood CD34+ cell concentration was >15-20 x 106/l, and the support variables in autologous haematopoietic stem cell collection target was > 2.5 x 106 CD34+ cells/kg (double for tan- transplantation dem ASCT). Mainly, if blood CD34+ cell count was <20 x 106/l P. Gómez Prieto, I. Rivas, M. Gasior, R. Guerrero, A. Marcos, and/or blood leukocyte count >20 x 109/l, plerixafor was added M.C. López, R. De Paz, A. López De La Guía, M. Canales, R. Arrieta to G-CSF. Hospital Universitario La Paz (Madrid, ES) Results: The results are shown in Table where blood CD34+ cell concentrations and the respective harvest yields are shown, either Targets: The study fi rst deals with the question of whether the after G-CSF only or after G-CSF + plerixafor. reduced use of Melphalan in the induction phase, has led to Conclusion: G-CSF only can successfully be used in mobilisation diminished transfusion requirements of both red blood cells (RBC) but additional plerixafor is often needed to get an adequate and platelets during the transplant phase. The study deals also yield - in 60 % of patients in our small pilot series. with the correlation between the amount of transfused RBC in the transplant, and the number of infused mononuclear cells (MNC). Additionally, the relationship between the number of CD34 cells administered and platelet consumption in autologous transplants is also investigated. Study Design and Methods: The sample group used for the fi rst study uses from 81 patients between 47 and 66 years old in the period 1996-2011, including the amount of both RBC and platelets administered during the transplant, patient’s volemia and if they had been previously treated with Melphalan in the pre-transplant induction phase (Group 1, n = 27) or not (Group 2, n = 54). The following formula is used to analyse each received transfusion: (Vol of RBC/platelet concentrate (ml) X No. of transfused units)/Patient Volemia. The RBC consumption/No. of administered mononuclear cells relationship is evaluated by comparing the RBC ratio: volume of infused RBC/patient volemia, on the same 81 patients. The Platelets/CD34 correlation is measured using the infused platelets volume/patient volemia ratio, on the same sample. Results: Our data found a signifi cant reduction on the average transfusion requirements for patients treated with Melphalan alternatives of both RBC and platelets (Table 1). The statistical test of the results, however, concludes that these results are not

S545 500/μl occurred on day +13. The patient was discharged at +15 day with an acceptable blood cell count. One month later the transplant the neurologic impairment was stable and a new immunoelectro- phoresis was made, and no monoclonal band has found. One year after the transplant the patient restore the muscular strength and the ability to walk without support. The last medical evaluation was two months ago, the patient was completely asymptomatic. The autologous stem cell transplantation is the fi rst choice of treatment in POEMS syndrome

Cellular and gene therapies and cytokines

R1547 Pre-emptive donor lymphocyte infusions after haploidentical stem cells transplantation favour long lasting remission in a case of advanced and refractory Hodgkin’s disease M. Dozzo, A. Sperotto, M. Battista, R. Fanin, F. Patriarca statistically signifi cant. Evidence of the infusion of MNC aff ecting Division of Haematology (Udine, IT) the RBC consumption during the transplant phase could not be found in the analysed data (Table 2). A correlation between lev- Background and purpose: Donor lymphocyte infusions (DLI) can els of CD34 cells and platelet requirements during the transplant potentially increase the immune-mediated anti-tumoral activ- phase could not be found either (Table 3). ity and provide many patients with an option of rescue, in case Conclusion: Although transfusion requirements of both RBC and of relapse after stem cell transplantation in terms of both disease platelets have been lower after Melphalan was replaced in induc- control and survival. Moreover, pre-emptive DLIs may favour a tion regimens, the result could not be deemed to be statistically long-lasting remission in patients at high risk of relapse after trans- signifi cant. No statistical proof of a correlation between the RBC plantation, but their use is limited due to the risk of acute GvHD, volumetric ratio during the transplant phase and the number of particularly after transplants with partially HLA matched donors. infused MNC was found on our data. Variations on the amounts of Patient: Here we report a patient with diagnosis of Hodgkin’s dis- administered CD34 cells could not be demonstrated by our data to ease, classical variant, nodular sclerosis, stage II A (mediastinum, have an eff ect on the transplant-phase platelet volumetric ratio. left pleural shedding), Hasenclever index 3, since September 2006, refractory to 4 lines of chemotherapy and mediastinal radiotherapy and progressed after autologous stem cell transplanta- R1546 tion (ASCT) performed on February 2009. He was submitted to an A successful autologous stem cell transplant for curative allogeneic SCT with clinically active disease (lymph nodes at both treatment of POEMS syndrome and a progression-free armpit areas grown, maximum diameter about 5 cm, and pleural survival of 49 months. A case report shedding) on 22-23th July 2010. Conditioning was with Treosul- M. Zapata, K. Galvez, F. Combariza fan and Fludarabine; GvHD prophylaxis was with ATG Fresenius, Hospital Pablo Tobon Uribe (Medellin, CO) Rapamycin and Mycophenolate. He received unmanipulated PBSC (11.3 x 106/kg of CD34+, 41.9 x 107/Kg cellule CD3+) from a We report about a 65 years old man previously asymptomatic. The 5/10 HLA- matched sibling donor. Engrafment took place on day patient was referred 2 years ago to the neurology service for a his- +14 with neutrophils more than 1000/mmc and on day +38 with tory of 1 year of progressive paresthesias in lower limbs to mid-leg platelets more than 50.000/mmc. and subsequently in the hands to the wrists and a progressive loss The main complication was toxic encephalopathy with disorien- of strength more in lower limbs. Besides this a loss of 24 kg in this tation and delirium of persecution since day +5 due to elevated period. The patient was evaluated with a central nervous MRI that serum levels of Rapamycin, that slowly resolved in the following was normal, Endocrinologist evaluation was made and confi rmed 2 weeks after Rapamycin withdrawal since day +47. At day +30 an autoimmune hypothyroidism, and a fasting glucose impairment the patient was in complete remission (CR) by clinical examina- (HbA1c 6mg/dl), fi nally was discarded another endocrine comint- tion and TC-PET, without signs of GvHD and with 100% donor chi- ment.. At the time of the neurologist evaluation the evidence of merism. After mycofenolate withdrawal at day +237 he received loss of muscle strength in upper and lower limbs were generalized 7 of escalating dose of DLI (0.5 x 105/Kg – 3 x 106/Kg) between arefl exia and hypoesthesia in boot and glove, and no sphincter day +370 and day +822. At the last follow-up, at day +813 after involvement, clubbing. It was considered as working diagnosis transplant, the patient is in continuous CR, without any sign of of polyneuropathy predominantly sensory axonal motor. Lum- chronic GvHD. bar puncture was performed and a new electromyography study Conclusion: This case-report suggests that haploidentical trans- performed. In cerebrospinal fl uid proteins were high with a low plant can be a salvage option for patients with advanced and cells count. A diagnosis of chronic infl ammatory polyneuropathy refractory Hodgkin disease and that a program of escalating is considered and secondary causes were sought. No evidence of doses of pre-emptive DLI is expected to be promising for favour- secondary disease was found but a serum electrophoresis showed ing long- lasting CR. a mild γ region monoclonal peak. The bone marrow study was indicated and report a normal hematopoiesis bone, reported a 4% plasma cells. The immuno electrophoresis reported a monoclonal R1548 IgG with a light Λ chains. And a long bones radiography evaluation Vaccine-induced anti-donor T-cell immunity for the with report of a multiple axial osteoesclerotic lesions. Based on the treatment of graft-versus-host disease after liver entire clinic the diagnosis of POEMS syndrome was made and treat- transplantation ment with a prednisone and cyclophosphamide was started while A. Shimabukuro-Vornhagen, S. Theurich, D. Stippel, an autologous hematopoietic progenitors transplant protocol was M. von Bergwelt-Baildon made. One month later Following a conditioning consisting of mel- University Hospital of Cologne (Cologne, DE) phalan based regimen (200mg/m2) an autologous bone marrow stem cells y two day separated infusion (total 2,23x106/kg CD34+) We report the case of a male patient who developed graft- was performed. Engraftment with absolute neutrophil counts > versus-host disease (GVHD) after orthotopic liver transplanation.

S546 Liver transplant-associated GVHD carries a poor prognosis with pre-treated with cream containing the TLR7 agonist imiquimod a high mortality rate and there are currently no standard treat- as an adjuvant. Unfortunately, the patient eventually died due to ment recommendations. Despite increased immunosuppressive severe sepsis. In order to assess the eff ectiveness of the vaccina- treatment the patients condition continued to deteriorate. We tions we quantifi ed the alloreactive T cell response of recipient thus reasoned that a reduction of the imunosuppressive ther- T cells against donor PBMCs using a mixed lymphocyte reaction apy could improve the GVHD by shifting the balance in favor of (MLR). We detected an increase in donor-reactive T lymphocytes recipient immunity and rejection of the donor lymphocytes. To after the vaccinations. These results indicate that our therapeutic further enhance anti-donor immunity we repeatedly vaccinated strategy was successful at inducing anti-donor immunity. Vaccina- the patient by injecting donor-derived peripheral blood mononu- tion against donor antigens could therefore represent a promis- clear cells (PBMC) intradermally into skin sites, which have been ing strategy for the treatment of GVHD after liver transplantation.

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