DOCSLIB.ORG

The Effects of Immunophenotyping with Flow Cytometry on Prognosis in Acute Lymphoblastic Leukemia

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Effects of Immunophenotyping with Flow Cytometry on Prognosis in Acute Lymphoblastic Leukemia JOURNAL OF Journal of Contemporary CONTEMPORARY MEDICINE Medicine DOI: 10.16899/jcm.787016 J Contemp Med 2021;11(1):22-28 Orjinal Araştırma / Original Article The Effects of Immunophenotyping with Flow Cytometry on Prognosis in Acute Lymphoblastic Leukemia Akut Lenfoblastik Lösemide Flow Sitometri ile İmmünofenotiplemenin Prognoz Üzerine Etkileri Sultan Aydin Köker1, Yeşim Oymak2, Dilek İnce3, Raziye Canan Vergin2, Ferah Genel4 1Department of Pediatric Hematology and Oncology, Manisa City Hospital, Manisa, Turkey 2Department of Pediatric Hematology and Oncology, Dr. Behçet Uz Children’s Hospital, Izmir, Turkey 3Department of Pediatric Oncology, Dokuz Eylül University School of Medicine, Izmir, Turkey 4Department of Pediatric Immunology, Dr. Behçet Uz Children’s Hospital, Izmir, Turkey Abstract Öz Background: The identification of immunophenotype subgroups Giriş: Akut lenfoblastik löseminin (ALL) tanı ve prognozu için is very important for the diagnosis and prognosis of acute immünofenotip alt gruplarının tanımlanması çok önemlidir. lymphoblastic leukemia (ALL). Gereç ve Yöntem: Çalışma, TR-ALL 2000 (modifiye) BFM tedavi Material and Method: The study is designed as a retrospective protokolü ile tedavi edilen 105 ALL olgu (65 erkek, 40 kadın; ortalama cohort study which included 105 children with ALL (65 males, 40 females; mean age 5.9±3.8 years) who were treated TR-ALL 2000 yaş 5.9 ± 3.8 yıl) içeren retrospektif kohort çalışması olarak tasarlanmıştı. (modified) BFM treatment protocol. Bulgular: EGIL sınıflaması dağılımları pro-B ALL (n=1), common B ALL Results: The distributions of EGIL classification were pro-B ALL (n=46), pre-B ALL (n=40), pre-T ALL (n=8), kortikal T ALL (n=6) ve matür (n=1), common B ALL (n=46), pre-B ALL (n=40), pre-T ALL (n=8), T ALL (n=4). T ALL grubunda lökosit ≥100.000/mm³, lenfadenopati cortical T ALL (n=6), and mature T ALL (n=4). Leukocyte≥100,000/ ≥2 cm, mediastinal tutulum yaygın olarak tespit edildi. T ALL, 8.gün mm³, lymphadenopathy≥2 cm, mediastinal involvement were periferik kan yaymasında blast sayısı ve 15. gün kemik iliği aspirasyonu commonly identified in T ALL group. T ALL had a poor response to chemotherapy according to 8th-day peripheral circulation (KİA)’ nundaki blast sayısına göre kemoterapiye kötü yanıt gösterdiği blast counts and 15th-day bone marrow aspiration (BMA) blast saptandı. Tedavinin indüksiyon döneminde nüks, mortalite ve ölüm counts. The recurrence, mortality, and death rate in the induction oranı T ALL grubunda sık olarak görüldü. Tek değişkenli analizle period of treatment were frequently detected in T ALL group. The gösterilen prognostik potansiyele sahip olan değişkenler tanı anında variables that had prognostic potential, as indicated by univariate lökosit sayısı, hepatomegali, splenomegali ve lenfadenopati, 8. gün analyses, were leukocyte count, hepatomegaly, splenomegaly, and lymphadenopathy at the time of diagnosis, 8th-day steroid steroid yanıtı, 15. gün KİA yanıtı, risk grubu, nüks ve immünofenotip response, 15th-day BMA response, risk group, recurrence, and idi. Çok değişkenli Cox regresyon analizinde ise sadece lökosit immunophenotyping. Multivariate Cox regression analysis sayısının (HR 2.51, p <0.001) prognozu önemli derecede etkilediğini demonstrated that only the leukocyte count (HR 2.51, p < 0.001) göstermiştir. was a predictor of prognosis. Sonuç: İmmünofenotipleme ALL'nin tanı ve prognozunda, risk Conclusion: Immunophenotyping may be effective in the gruplarının tanımlanmasında ve riske baglı tedavi planlamasında etkili diagnosis and prognosis of ALL, identification of risk groups, and in risk-based treatment planning. T ALL had a poor prognosis. olabilir. T ALL tanısı alan hastalarda prognozun kötü olacağı gösterildi. Keywords: Flow cytometry, immunophenotyping, acute Anahtar Kelimeler: Akım sitometrisi, immünofenotipleme, akut lymphoblastic leukemia, children lenfoblastik lösemi, çocuk Corresponding (İletişim): Sultan Aydin Köker, Department of Pediatric Hematology and Oncology, Manisa City Hospital, Manisa, Turkey E-mail (E-posta): [email protected] Received (Geliş Tarihi): 28.08.2020 Accepted (Kabul Tarihi): 02.11.2020 23 Journal of Contemporary Medicine INTRODUCTION quadrant on physical examination. Testis involvement was Leukemia comprises nearly 25-30% of childhood malignancies.1 evaluated by testis ultrasonography for all male patients. For acute lymphoblastic leukemia (ALL), previously known as a Cytogenetic, FISH and molecular investigations examined deadly disease, developments in diagnosis and treatment and in aspiration material and t (9;22), t (1;19), t (4;11), and t advances in support treatments have achieved lengthened (12;21) translocations were evaluated at the Gentest Genetic disease-free survival in most cases. In fact, 70-80% of cases Laboratory in Özel Şifa Hospital. have the chance of full recovery from the disease (cure).1,2 Prednisone response is explained as the peripheral blast count For pediatric ALL patients, for determination of both risk on day 8 of treatment. Prednisone good response (PGR) was group and prognosis, the following factors are known; age, characterized by <1,000/μL, while prednisone poor response gender, leukocyte count, hematocrit value, platelet count, (PPR) showed ≥1,000/μL peripheral blast count on 8th- hepatomegaly, splenomegaly, lymphadenopathy presence, day treatment. Response in BM was evaluated on 15th-day MSS involvement, testis involvement, mediastinal mass, and 33th-day of induction treatment and was classified as and other extramedullary involvement, French-American- M1 (<5%), M2 (5 to <25%), and M3 (≥25% leukemic blasts). British (FAB) classification (L1, L2, L3), response to treatment, Complete remission (CR) was described as M1 BM on day the molecular genetics such translocations, and the Minimal 33 of induction therapy, the absence of leukemic blasts in Residual Disease (MRD) status is defined by qPCR or NGS. blood and CSF, and no symptom of local disease. Relapse was Additionally, in recent times immunophenotyping results characterized by recurrence of 25% lymphoblasts or over in BM obtained by investigation of cytogenetic and molecular genetic or local leukemic involvements at any site. disorders with flow cytometry have gained importance.3-6 Risk group stratification is categorized three groups; patients The study aims to research the properties and effect on with initial leukocyte count <20,000/mm3 and age ≥1 and < the prognosis of ALL immunophenotype subgroups and 6 years, and blast counts <1,000/mm3 in peripheral blood on the evaluation of correlations between clinical and other 8th-day after 7 day prednisolone and IT MTX treatment, no T cell laboratory data identified during the diagnosis stage. immunology, t (9;22) (BCR/ABL) and t (4;11) (MLL/AF4) negative and M1 on 33th-day were defined as standard risk group (SRG). The patients with initial leukocyte count ≥ 20,000/mm3 or age MATERIAL AND METHOD <1 or ≥6 years, and blast counts <1,000/mm3 in peripheral The study included 105 cases with ALL diagnosed in Pediatric blood on 8th-day after 7 day prednisolone and IT MTX treatment, Hematology and Oncology Department and treated with TR- t (9;22) (BCR/ABL) and t (4;11) (MLL/AF4) negative on 33th-day, ALL 2000 (modified) BFM protocol, followed up between January were described as moderate risk group (MRG). 2008 to March 2012. The study is designed as retrospective The cases with blast counts ≥1000/mm3 on 8th-day in cohort study. The patients’ gender, age at diagnosis, leukocyte peripheral blood, t (9;22) (BCR/ABL) and t (4;11) (MLL/AF4) count, hematocrit value, platelet count, hepatomegaly, positive, without full remission on 33th-day, with hypodiploidy splenomegaly, lymphadenomegaly, central nervous system (chromosome <45) chromosome anomaly, and presence of (CNS) involvement, testis involvement, mediastinal mass each criteria alone independent of age and leukocyte count and other extramedullary involvement, French-American- (one criterion is sufficient) were characterized as high risk British (FAB) classification (L1, L2, L3), immunophenotyping group (HRG).6 with flow cytometric investigation, translocations (t (4;11), t (9;22), t (12,21), t (1;19)), 8th-day prednisone response, and Flow cytometry 15th-day and 33th-day bone marrow aspiration (BMA) blast Immunophenotyping assays were administered by four‐color rates, risk groups, surveillance duration, relaps, and mortality [FITC (Fluorescein Isothiocyanate), PE (Phycoerythrin), PC5 were retrospectively obtained from patients’ medical records. (Phycoerythrin-Cyanine 5) or PC7 (Phycoerythrin-Cyanine 7)] The study was approved by the local Ethics Committee of Dr. flow cytometry analysis of BM samples. Cytoplasmic and surface Behçet Uz Children’s Hospital in accordance with the Helsinki cell markers were identified using according to Immunological Declaration (Project Number: 2012/47). Informed consent was Characterization of Leukemia (EGIL) classification.7 obtained from each parent and/or patients. The reactivity with fluorescent‐conjugated monoclonal Mediastinal involvement was assessed by thorax radiography antibodies (Moab) was against T‐cell‐related antigens (CD1a, (P/A and lateral). Central nervous system (CNS) involvement CD2, CD3, CD4, CD5, CD7, and CD8), B‐cell differentiation was defined as presence of leukemic cells in cerebrospinal markers (CD10, CD19, CD20, CD22, CD79a), myeloid cell fluid (CSF) with lumbar puncture and findings by Magnetic antigens (CD11b, CD13, CD14, CD15, CD33, CD117, and Resonance Imaging (MRI).
Load more

Recommended publications
  • Bone Marrow Immunophenotyping by Flow Cytometry in Refractory Cytopenia of Childhood
    Myelodysplastic Syndromes ARTICLES Bone marrow immunophenotyping by flow cytometry in refractory cytopenia of childhood Anna M. Aalbers,1,2 Marry M. van den Heuvel-Eibrink,2 Irith Baumann,3 Michael Dworzak,4 Henrik Hasle,5 Franco Locatelli,6 Barbara De Moerloose,7 Markus Schmugge,8 Ester Mejstrikova,9 Michaela Nováková,9 Marco Zecca,10 C. Michel Zwaan,2 Jeroen G. te Marvelde,1 Anton W. Langerak,1 Jacques J.M. van Dongen,1 Rob Pieters,2 Charlotte M. Niemeyer,11 and Vincent H.J. van der Velden1 1Department of Immunology, Erasmus MC, Erasmus University Medical Center, Rotterdam, The Netherlands; 2Department of Pediatric Oncology/Hematology, Sophia Children’s Hospital - Erasmus University Medical Center, Rotterdam, The Netherlands; 3Department of Pathology, Clinical Centre South West, Böblingen Clinics, Germany; 4St. Anna Children’s Hospital and Children’s Cancer Research Institute, Department of Pediatrics, Medical University of Vienna, Austria; 5Department of Pediatrics, Aarhus University Hospital Skejby, Aarhus, Denmark; 6Department of Pediatric Hematology-Oncology, IRCCS Ospedale Bambino Gesù, Rome, University of Pavia, Italy; 7Department of Pediatric Hematology/Oncology, Ghent University Hospital, Ghent, Belgium; 8Department of Hematology, University Children’s Hospital, Zurich, Switzerland; 9Department of Pediatric Hematology/Oncology, Charles University and University Hospital Motol, Prague, Czech Republic; 10Pediatric Hematology, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy; and 11Department of Pediatrics and Adolescent Medicine, Division of Pediatric Hematology and Oncology, University of Freiburg, Germany ABSTRACT Refractory cytopenia of childhood is the most common type of childhood myelodysplastic syndrome. Because the majority of children with refractory cytopenia have a normal karyotype and a hypocellular bone marrow, differ- entiating refractory cytopenia from the immune-mediated bone marrow failure syndrome (very) severe aplastic anemia can be challenging.
    [Show full text]
  • The AML Guide Information for Patients and Caregivers Acute Myeloid Leukemia
    The AML Guide Information for Patients and Caregivers Acute Myeloid Leukemia Emily, AML survivor Revised 2012 Inside Front Cover A Message from Louis J. DeGennaro, PhD President and CEO of The Leukemia & Lymphoma Society The Leukemia & Lymphoma Society (LLS) wants to bring you the most up-to-date blood cancer information. We know how important it is for you to understand your treatment and support options. With this knowledge, you can work with members of your healthcare team to move forward with the hope of remission and recovery. Our vision is that one day most people who have been diagnosed with acute myeloid leukemia (AML) will be cured or will be able to manage their disease and have a good quality of life. We hope that the information in this Guide will help you along your journey. LLS is the world’s largest voluntary health organization dedicated to funding blood cancer research, advocacy and patient services. Since the first funding in 1954, LLS has invested more than $814 million in research specifically targeting blood cancers. We will continue to invest in research for cures and in programs and services that improve the quality of life for people who have AML and their families. We wish you well. Louis J. DeGennaro, PhD President and Chief Executive Officer The Leukemia & Lymphoma Society Inside This Guide 2 Introduction 3 Here to Help 6 Part 1—Understanding AML About Marrow, Blood and Blood Cells About AML Diagnosis Types of AML 11 Part 2—Treatment Choosing a Specialist Ask Your Doctor Treatment Planning About AML Treatments Relapsed or Refractory AML Stem Cell Transplantation Acute Promyelocytic Leukemia (APL) Treatment Acute Monocytic Leukemia Treatment AML Treatment in Children AML Treatment in Older Patients 24 Part 3—About Clinical Trials 25 Part 4—Side Effects and Follow-Up Care Side Effects of AML Treatment Long-Term and Late Effects Follow-up Care Tracking Your AML Tests 30 Take Care of Yourself 31 Medical Terms This LLS Guide about AML is for information only.
    [Show full text]
  • Treatment Induced Cytotoxic T-Cell Modulation in Multiple Myeloma Patients
    ORIGINAL RESEARCH published: 15 June 2021 doi: 10.3389/fonc.2021.682658 Treatment Induced Cytotoxic T-Cell Modulation in Multiple Myeloma Patients Gregorio Barilà, Laura Pavan, Susanna Vedovato, Tamara Berno, Mariella Lo Schirico, Massimiliano Arangio Febbo, Antonella Teramo, Giulia Calabretto, Cristina Vicenzetto, Vanessa Rebecca Gasparini, Anna Fregnani, Sabrina Manni, Valentina Trimarco, Samuela Carraro, Monica Facco, Francesco Piazza, Gianpietro Semenzato and Renato Zambello* Department of Medicine (DIMED), Hematology and Clinical Immunology Section, Padua University School of Medicine, Edited by: Padova, Italy Roberto Mina, Università degli Studi di Torino, Italy Reviewed by: The biology of plasma cell dyscrasias (PCD) involves both genetic and immune-related Mattia D’ Agostino, factors. Since genetic lesions are necessary but not sufficient for Multiple Myeloma (MM) University of Turin, Italy evolution, several authors hypothesized that immune dysfunction involving both B and T Michele Cea, University of Genoa, Italy cell counterparts plays a key role in the pathogenesis of the disease. The aim of this study *Correspondence: is to evaluate the impact of cornerstone treatments for Multiple Myeloma into immune Renato Zambello system shaping. A large series of 976 bone marrow samples from 735 patients affected by [email protected] PCD was studied by flow analysis to identify discrete immune subsets. Treated MM Specialty section: samples displayed a reduction of CD4+ cells (p<0.0001) and an increase of CD8+ This article was submitted to (p<0.0001), CD8+/DR+ (p<0.0001) and CD3+/CD57+ (p<0.0001) cells. Although these Hematologic Malignancies, findings were to some extent demonstrated also following bortezomib treatment, a more a section of the journal Frontiers in Oncology pronounced cytotoxic polarization was shown after exposure to autologous stem cell Received: 18 March 2021 transplantation (ASCT) and Lenalidomide (Len) treatment.
    [Show full text]
  • Current Challenges in Providing Good Leukapheresis Products for Manufacturing of CAR-T Cells for Patients with Relapsed/Refractory NHL Or ALL
    cells Article Current Challenges in Providing Good Leukapheresis Products for Manufacturing of CAR-T Cells for Patients with Relapsed/Refractory NHL or ALL Felix Korell 1,*, Sascha Laier 2, Sandra Sauer 1, Kaya Veelken 1, Hannah Hennemann 1, Maria-Luisa Schubert 1, Tim Sauer 1, Petra Pavel 2, Carsten Mueller-Tidow 1, Peter Dreger 1, Michael Schmitt 1 and Anita Schmitt 1 1 Department of Internal Medicine V, University Hospital Heidelberg, 69120 Heidelberg, Germany; [email protected] (S.S.); [email protected] (K.V.); [email protected] (H.H.); [email protected] (M.-L.S.); [email protected] (T.S.); [email protected] (C.M.-T.); [email protected] (P.D.); [email protected] (M.S.); [email protected] (A.S.) 2 Institute of Clinical Transfusion Medicine and Cell Therapy (IKTZ), 89081 Heidelberg, Germany; [email protected] (S.L.); [email protected] (P.P.) * Correspondence: [email protected] Received: 9 April 2020; Accepted: 13 May 2020; Published: 15 May 2020 Abstract: Background: T lymphocyte collection through leukapheresis is an essential step for chimeric antigen receptor T (CAR-T) cell therapy. Timing of apheresis is challenging in heavily pretreated patients who suffer from rapid progressive disease and receive T cell impairing medication. Methods: A total of 75 unstimulated leukaphereses were analyzed including 45 aphereses in patients and 30 in healthy donors. Thereof, 41 adult patients with Non-Hodgkin’s lymphoma (85%) or acute lymphoblastic leukemia (15%) underwent leukapheresis for CAR-T cell production.
    [Show full text]
  • Characterization of Immunophenotyping in ALL: a Single Center Study of Western India Dr
    DOI: 10.21276/sjams.2016.4.9.35 Scholars Journal of Applied Medical Sciences (SJAMS) ISSN 2320-6691 (Online) Sch. J. App. Med. Sci., 2016; 4(9C):3349-3354 ISSN 2347-954X (Print) ©Scholars Academic and Scientific Publisher (An International Publisher for Academic and Scientific Resources) www.saspublisher.com Original Research Article Characterization of Immunophenotyping in ALL: A Single Center Study of Western India Dr. Sandeeep K Jasuja1, Dr Sandhya Gulati2, Shraddha Patel3, Dr Nidhi Sharma4 1Asso. Professor and Head, Department of Medical Oncology, SMS MC & Attached Hospitals, Jaipur, Rajasthan, India 2Professor, Department of Pathology, SMS MC & Attached Hospitals, Jaipur, Rajasthan, India 3Observational Trainer, Department of Medical Oncology, SMS MC & Attached Hospitals, Jaipur, Rajasthan, India 4Assistant Professor, Department of Pathology, SMS MC & Attached Hospitals, Jaipur, Rajasthan, India *Corresponding author Dr. Sandeeep K Jasuja Email: [email protected] Abstract: This retrospective observational study was done among 227 ALL cases at a cancer treatment center in Western India. Data were collected retrospectively from hospital records during June 2015 to May 2016. Diagnosis of ALL was made based on the complete blood cell counts, peripheral blood smear/bone marrow aspirate smear and immunophenotyping study. Out of 227 ALL patients, 217 cases were of B-ALL, 8 cases were of T-ALL and 2 cases were of biphenotypic ALL. Of these 217 cases of B-ALL 190 (87.56) were pediatric age (range 0.5-15 years) and 27 (12.44) were adult (range 16-52 years). T-ALL cases were equally distributed in both age groups, 4 (50%) cases in pediatric age group (range 1-7 years)and 4 (50%) cases were adult (range 19-62 years).
    [Show full text]
  • Plasma-Cell-Dyscrasias-Sflc-Assay Executive
    Comparative Effectiveness Review Number 73 Effective Health Care Program Serum Free Light Chain Analysis for the Diagnosis, Management, and Prognosis of Plasma Cell Dyscrasias Executive Summary Background Effective Health Care Program Plasma cell dyscrasias (PCDs) are a group The Effective Health Care Program of neoplastic disorders characterized by was initiated in 2005 to provide the uninhibited expansion of a monoclonal valid evidence about the comparative population of malignant plasma cells.1 effectiveness of different medical Multiple myeloma (MM) is the most interventions. The object is to help common malignant plasma cell tumor, consumers, health care providers, accounting for about 1 percent of all and others in making informed cancer types,1 and the second most choices among treatment alternatives. common hematologic malignancy in Through its Comparative Effectiveness the United States. With an age-adjusted Reviews, the program supports incidence rate of 5.5 cases per 100,000 systematic appraisals of existing population,2 an estimated 19,900 new scientific evidence regarding diagnoses and 10,790 deaths due to treatments for high-priority health myeloma occurred in 2007, according conditions. It also promotes and to the American Cancer Society.3 Although generates new scientific evidence by the median survival has improved to identifying gaps in existing scientific 5 years with current standards of evidence and supporting new research. treatment,4 the annual costs of modern The program puts special emphasis therapies can range from $50,000 to on translating findings into a variety $125,000 per patient.5,6 of useful formats for different In PCDs, each abnormally expanded stakeholders, including consumers.
    [Show full text]
  • Immunophenotyping of Acute Leukaemias Immunophänotypisierung Akuter Leukämien
    Immunhämatologie Redaktion: G. Rothe Immunophenotyping of Acute Leukaemias Immunophänotypisierung akuter Leukämien T. Benter, R. Rätei, W.-D. Ludwig Summary: For nearly 100 years the classification of menden Einfluß gewonnen. Der Grund liegt in den blood cells and the diagnosis of leukaemia have been Fortschritten der Laser- und Computertechnologie, based on cytomorphological features after staining. aber auch an der Verfügbarkeit von Hunderten ver- Even in the era of molecular biology this is still es- schiedener monoklonaler Antikörper (moAB). die sential. Therapy of acute myeloid leukaemia (AML) is gegen eine Vielfalt von Antigenen hämatopoetischer mostly dependent on the interpretation of the morpho- Zellen gerichtet sind. logical appearance of blasts under the microscope. Cy- Dieser Übersichtsartikel fokussiert auf die Immun- tomorphology should also lead to a rational use of phänotypisierung von Patienten mit akuten Leukämien techniques like immunophenotyping, cytogenetics, flu- und zeigt den Einfluß auf die Diagnostik und Thera- orescence / situ hybridisation (FISH), and poly- pie. merase chain reaction (PCR). In the past two decades, the impact of immunophe- Schlüsselwörter: akute Leukämie: Immunophänoty- notyping by flow cytometry in the diagnosis and man- pisierung: Klassifikation von Leukämien. agement of acute leukaemia has expanded rapidly. This has been mainly attributed to significant advances in laser and computer technologies and the production of several hundred monoclonal antibodies (moAbs) to he gold standard for classifying acute myeloid a variety of antinens expressed by haematopoietic Tleukaemia (AML) has been based on morphologi- cells. cal, cytochemical, and iinmunophenotypic criteria as This review concentrates on immunophenotyping of defined by the French—American—British (FAB) sys- cells from patients with acute leukaemia and shows the tem 11-4].
    [Show full text]
  • Immunophenotyping of B-Acute Lymphoblastic Leukaemia
    IMMUNOPHENOTYPING OF B-ACUTE LYMPHOBLASTIC LEUKAEMIA Dr Lee Shir Ying Sr Consultant Haematologist Department of Laboratory Medicine National University Hospital Singapore Introduction • Case study illustrating the principles of immunophenotyping for B-ALL • MRD detection in B-ALL Cell characterization • Means of characterization: – Morphology, cytochemistry, immunohistochemistry, flow cytometry immunophenotyping – Characterization into: –Lineage, maturity, abnormality Flow Cytometry Technology • Simultaneous measurement of multiple physical characteristics of a single cell as the cell flows in suspension through a measuring device Definition • B-ALL: Neoplasm of precursors (lymphoblasts) committed to B-cell lineage • When blood and bone marrow are extensively involved, acute lymphoblastic leukemia (B-ALL) is appropriate • When disease is confined to mass with absent or minimal blood and marrow involvement, term lymphoblastic lymphoma (B-LBL) is used • If patient presents with mass lesion and blood and bone marrow involvement, 25% blasts in marrow defines leukemia (B-ALL) Case Study • 58-year-old man. No past medical history of note. • Referred for investigation of pancytopaenia and constitutional symptoms. • Also a paravertebral mass, which was biopsied (also sent for flow cytometry) • Full blood count – Total white cell count - 2.9 x 109/L – Haemoglobin - 7.7 g/dL – Platelet count - 23 x 109/L ➢ Bone marrow aspirate MGG stained ➢ Infiltrated by small uniform blast-like cells ➢ Fine chromatin, high N/C ratio, nucleoli present, irregular nuclear contours Bone marrow trephine: ➢ Hypercellular for age ➢ Marked decrease in trilineage haematopoiesis Sheets of darkly-stained cells with high N/C • Round to irregular nuclei • Evenly dispersed chromatin • Inconspicuous to small nucleoli PAX5 CD79a CD20 Morphology appears to be a straightforward B-cell acute lymphoblastic leukaemia, but....
    [Show full text]
  • Clearllab 10C Panel Markers and How They Are Combined
    WHITE PAPER ClearLLab 10C Panel Markers and how they are combined Mike Keeney London Health Sciences Centre, Victoria Hospital, London, ON, Canada Sandra Hernandez Beckman Coulter Inc. Life Sciences, Miami, FL 33196 IN THIS PAPER YOU WILL Experience the utility of the Discover the ClearLLab 10C Explore the power of optimizing ClearLLab 10C panel as an aid panel: the first 10-color CE-IVD your L&L immunophenotyping in the differential diagnosis of immunophenotyping reagents workflow with fewer steps, hematologically abnormal patients for assessing lymphoid and fewer tubes and more colors having or suspected of having myeloid populations certain hematopoietic neoplasms Introduction Flow cytometric immunophenotyping remains an indispensable tool for the diagnosis, classification, staging, and monitoring of hematologic neoplasms. There have been significant advances in flow cytometry instrumentation and availability of an expanded range of antibodies and fluorochromes that have improved the ability to identify different normal cell populations and recognize phenotypic aberrancies. ClearLLab 10C Panels consists of a set of four 10-Color reagents in dry unitized format, composed of antibodies directed against T, B, NK and Myeloid lineage antigens. These panels are designed to include the primary antibodies which are lineage oriented as per the 2006 Bethesda consensus. The combinations are intended to identify the leukocyte subpopulations that express these antigens alone or in specific co-expression patterns, and are designed to work with both normal samples and samples coming from any patient having or suspected of having the following hematopoietic neoplasms: chronic leukemia, acute leukemia, non-Hodgkin lymphoma, myeloma, myelodysplastic syndrome (MDS), and/or myeloproliferative neoplasms (MPN).
    [Show full text]
  • Correlation Between Flow Cytometry and Histologic Findings
    ORIGINAL ARTICLE Correlation between flow cytometry and histologic findings: ten year experience in the investigation of lymphoproliferative diseases Correlação entre citometria de fluxo e anatomia patológica: experiência de dez anos na investigação de doenças linfoproliferativas Alanna Mara Pinheiro Sobreira Bezerra1, Denise da Cunha Pasqualin1, João Carlos de Campos Guerra2, Marjorie Paris Colombini3, Elvira Deolinda Rodrigues Pereira Velloso2, Paulo Augusto Achucarro Silveira2, Cristovão Luis Pitangueira Mangueira1, Ruth Hissae Kanayama3, Sonia Tsukasa Nozawa3, Rodolfo Correia3, Ana Carolina Apelle1, Welbert de Oliveira Pereira3, Rodrigo Gobbo Garcia4, Nydia Strachman Bacal5 ABSTRACT follicular lymphoma; 15 patients with diffuse large B-cell lymphoma; 7 Objective: To demonstrate the advantages of correlating patients with small lymphocytic B-cell lymphoma/chronic lymphocytic flow cytometry immunophenotyping with the pathology/ leukemia; 3 patients with mantle cell lymphoma; 1 patient with immunohistochemistry of lymph nodes or nodules in the diagnosis Burkitt’s lymphoma; 1 patient with MALT type lymphoma; 1 patient of lymphoproliferative diseases. Methods: A retrospective study with post-transplant lymphoproliferative disease; 2 patients with high was carried out of 157 biopsy or fine-needle aspiration lymph nodes/ grade non-Hodgkin’s B-cell lymphoma; 1 patient with low grade non- nodule specimens taken from 142 patients, from 1999 and 2009. Hodgkin’s B-cell lymphoma not otherwise specified; 1 patient with The specimens were simultaneously studied with flow cytometry Hodgkin’s lymphoma; and 12 patients with B-cell non-Hodgkin’s and pathology at Hospital Israelita Albert Einstein. The specimens lymphoma not otherwise specified. Conclusion: Flow cytometry were prepared in hematoxylin/eosin, Giemsa, or monoclonal antibody adds to the results of morphologic and immunohistochemical studies, stained slides for detecting specific antibodies for the purposes facilitating a rapid and accurate diagnosis of lymphoproliferative of pathology/immunohistochemical analysis.
    [Show full text]
  • Flow Cytometry CD45 Gating for Immunophenotyping of Acute
    Leukemia (1997) 11, 1878–1886 1997 Stockton Press All rights reserved 0887-6924/97 $12.00 Flow cytometry CD45 gating for immunophenotyping of acute myeloid leukemia F Lacombe, F Durrieu, A Briais, P Dumain, F Belloc, E Bascans, J Reiffers, MR Boisseau and Ph Bernard Laboratoire d’He´matologie, Service des Maladies du Sang, Hoˆpital du Haut-Le´veˆque, Pessac, France A flow cytometry method has been introduced into the routine age values within the malignant blast cell populations or investigation of whole bone marrow samples following red within the total nucleated cells present in each sample. In blood cell lysis on the basis of a primary CD45/side scatter (SSC) gating procedure. Blast cells were first identified by order to give numbers of malignant cells and proportions of CD45/SSC gating in 74 cases of acute myeloid leukemia (AML) cells within this cohort, a reliable practical discrimination and the results were compared to a conventional FSC/SSC gat- between malignant blasts and normal cell types would be ing procedure and to MGG-staining smears. The percentages required. In this paper, as previously recommended by of blast cells in these samples as defined by the morphological Borowitz et al,6 we suggest that such a discrimination can be analysis of MGG smears correlated better with the values readily facilitated by introducing primary gating for CD45 determined by CD45/SSC gating (r 5 0.94) than with the blast antigen expression and side scatter (CD45/SSC). We also sug- cell counts recorded with FSC/SSC gating (r 5 0.76). These findings were not surprising because while CD45 expression gest that this step should replace the first gating step for for- was regularly lower on leukemic blasts than on normal lymph- ward scatter and side scatter (FSC/SSC), as this latter procedure oid and monocytic cells, the FCS/SSC characteristics of these does not discriminate well between leukemic blasts, lympho- populations were overlapping.
    [Show full text]
  • Development of a Regional Flow Cytometry Center for Diagnosis Of
    Leukemia (2005) 19, 323–325 & 2005 Nature Publishing Group All rights reserved 0887-6924/05 $30.00 www.nature.com/leu EDITORIAL Development of a regional flow cytometry center for diagnosis of childhood leukemia in Central America SC Howard1,2,3, D Campana1,2,3,7, E Coustan-Smith1, FG Antillon3,4, M Bonilla3,5,LFu3,6, C-H Pui1,2,3,7, RC Ribeiro1,2,3, JA Wilimas1,2,3 and R Lorenzana8 1Department of Hematology-Oncology, St Jude Children’s Research Hospital, Memphis, TN, USA; 2Department of Pediatrics, University of Tennessee College of Medicine, Memphis, TN, USA; 3International Outreach Program, St Jude Children’s Research Hospital, Memphis, TN, USA; 4Unidad Nacional de Oncologı´a Pedia´trica, Guatemala City, Guatemala; 5Hospital Benjamin Bloom, San Salvador, El Salvador; 6Hospital Escuela, Tegucigalpa, Honduras; 7Department of Pathology, St Jude Children’s Research Hospital, Memphis, TN, USA; and 8Laboratorio de Patologı´a y Citometrı´a de Flujo, Guatemala City, Guatemala Leukemia (2005) 19, 323–325. doi:10.1038/sj.leu.2403624 From April 30, 1998 to December 31, 2002, specimens from pediatric patients with suspected leukemia at the Hospital Assessment of immunophenotype and DNA content of leukemia Escuela (Tegucigalpa, Honduras), the Hospital Benjamin Bloom is important for diagnosis and risk assessment, but requires (San Salvador, El Salvador), and the Unidad Nacional de costly equipment, reagents and technical expertise. Samples Oncologı´a Pedia´trica (Guatemala City, Guatemala) were from 1118 children in Central America were sent for flow analyzed by the Pathology and Flow Cytometry Laboratory in cytometric examination to a reference laboratory in Guatemala Guatemala City.
    [Show full text]