Acta Scientific Microbiology (ASMI) (ISSN: 2581-3226)

Research Article Volume 4 Issue 1

Immunophenotypic Profile of Patients with Clinically Diagnosed Chronic Lymphocytic Leukemia

Shirin Tarafder*

Professor, Department of Microbiology and Immunology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh

*Corresponding Author: Shirin Tarafder, Professor, Department of Microbiology and Immunology, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh.

Received: November 09, 2020; Published: December 28, 2020

×

Abstract

Background: The diagnosis of Chronic Lymphocytic Leukemia (CLL) is based on cell morphology, clinical manifestations and immunophenotyping. Immunophenotyping by Flow cytometry is the most reliable methodology for the diagnosis of CLL which is a clonal mature B-cell neoplasm (MBCN). This study aims to evaluate the application of multiparametric Flow Cytometry Immunophenotyping (FCI) as a standard methodology for the confirmation of or exclusion of CLL diagnosis in clinically suspected CLL patients. Methods: Four color FCI method was used in EDTA peripheral blood samples taken from 50 patients diagnosed preliminary as CLL through clinical data, complete blood count, peripheral blood film and bone marrow examination. The following fluorescent monoclonal antibodies were used: CD19, CD5, CD20, CD22, CD23, CD79b, FMC7, Kappa and Lambda light chains, CD200, CD123, CD10, CD11c, CD3, CD7, CD25, CD30, CD56, CD95, BCL2, CD34. FCI analysis was performed on a Beckman coulter Cytomics FC500 flow cytometer using software CXP to analyze data.

Result: Among 50 patients, 8(16%) showed normal T-cell population, while 42(84%) showed pathological B-cell lines. From these, 32 (64%) of 42 patients expressed typical CLL markers, whilst 10 (20%) of them showed B-cell prolymphocytic leukemia (B-PLL) profile. Scoring system proposed by Matutes., et al. and Moreau., et al. using CD5, CD23, CD22, FMC7, CD79b and SmIg (CLL, score 4-5 and other MBCN, score ≤ 3) differentiated CLL from B-PLL. CD200 was expressed in all (100%) CLL but not in B-PLL cases except 2(20%). Sensitivity and specificity of CD200 was 100% and 90% respectively.

Conclusion: FCI is a fundamental laboratory method without which final diagnosis of CLL can’t be established. CD200 expression is pivotal to MBCN classification. Expression of CD200 in CLL and other MBCNs, supports inclusion of this marker in flow cytometric panels for the differential diagnosis of CLL.

Keywords: Chronic Lymphocytic Leukemia; Flowcytometry; Immunophenotyping; Monoclonal Antibodies; Mature B-cell Neoplasms; CD200

×

References

  1. Jaffe ES., et al. “World Health Organization Classification of Tumours: Pathology and Genetics of Tumours of Haematopoietic and Lymphoid Tissues”. Lyon, France: IARC Press (2001): 127-130.
  2. Foucar K. “Bone marrow pathology”. In: Chronic lymphoproliferative disorders. Chicago: ASCP Press (1995).
  3. Huang JC., et al. “CD5-negative B-cell leukemias are rarely classifiable as chronic lymphocytic leukemia”. American Journal of Clinical Pathology 111 (1999): 123-130.
  4. Chiorazzi N., et al. “Chronic Lymphocytic Leukemia”. The New England Journal of Medicine 352 (2005): 804-815.
  5. Swerdlow SH., et al. “World Health Organization Classification of Tumours of Haematopoietic and Lymphoid Tissues”. Lyon: IARC Press (2008): 180-182.
  6. Hallek M., et al. “Guidelines for the diagnosis and treatment of chronic Lymphocytic leukemia: a report from the international workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute- Working group 1996 guidelines”. Blood 111 (2008): 5446-5456.
  7. Lin TS., et al. “Chronic Lymphocytic Leukemia”. In: Hoffman R, Benz. Jr EJ, Shattil SJ, Furie B, Silberstein LE, McGlave P, et al, editors. Hematology: Basic Principles and Practice. 5th ed. Churchill livingstone/Elsevier Philadelphia, USA (2008): Ch.83.
  8. Gribben JG. “Molecular profiling in CLL”. Hematology: the American Society of Hematology Education Program (2008): 444-449.
  9. Melo JV., et al. “The relationship between chronic lymphocytic leukemia and prolymphocytic leukemia: IV. Analysis of survival and prognostic features”. British Journal of Haematology 63 (1986): 377-387.
  10. Bennet JM., et al. “Proposal for the classification of chronic (mature) B and T lymphoid leukemias”. Journal of Clinical Pathology 42 (1989): 567-584.
  11. Bain BJ. “Blood cells. A Practical Guide”. Blackwell Publishing Limited. (2006): 439.
  12. Rai KR., et al. “Clinical staging of chronic lymphocytic leukemia”. Blood 46 (1975): 219-234.
  13. Rai KR. “A critical analysis of staging in CLL”. In: Gale RP, Rai KR, editors. Chronic Lymphocytic Leukemia: Recent Progress and Future Directions. New York, NY: Liss (1987): 253-264.
  14. Marti G., et al. “Diagnostic criteria for monoclonal B-cell lymphocytosis”. British Journal of Haematology 130 (2005): 325-332.
  15. D’Archangelo M. “Flow cytometry: new guidelines to support its clinical application Enumeration of Immunologically Defined Cell Populations by Flow Cytometry Approved Guideline, 2nd ed”. Cytometry 72B (2007): 209-10.
  16. Muller Hermelink HK., et al. “Chronic lymphocytic leukemia/small lymphocytic lymphoma”. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, et al., editors. World Health Organization Classification of Tumours of Haematopoietic and Lymphoid Tissues. Lyon: IARC Press (2008): 180-182.
  17. Matutes E., et al. “Diagnostic issues in chronic lymphocytic leukemia (CLL)”. Best Practice and Research Clinical Haematology 23 (2010): 3-20.
  18. Habib Nk and Finn WG. “Unsupervised immunophenotypic profiling of chronic lymphocytic leukemia”. Cytometry Part B: Clinical Cytometry 70 (2006): 124-135.
  19. Orfao A., et al. “Diagnosis of hematological malignancies: new applications for flow cytometry”. Hematology (EHA) 2 (2006): 6-13.
  20. Matutes E., et al. “The immunological profile of B-cell disorders and proposal of a scoring system for the diagnosis of CLL”. Leukemia 8 (1994): 1640-1645.
  21. Moreau EJ., et al. “Improvement of the chronic lymphocytic leukemia scoring system with the monoclonal antibody SN8 (CD79b)”. American Journal of Clinical Pathology 108 (1997): 378-382.
  22. Bene MC., et al. “Immunophenotyping of acute leukemia and lymphoproliferative disorders: a consensus proposal of the European Leukemia Net Work Package10”. Leukemia 25 (2011): 567-574.
  23. Ginaldi L., et al. “Levels of expression of CD19 and CD20 in chronic B cell leukaemias”. Journal of Clinical Pathology 51 (1998): 364-369.
  24. Chorazzi N., et al. “Cellular origin (s) of chronic lymphocytic leukemia: cautionary notes and additional considerations and possibilities”. Blood 117 (2011): 1781-1791.
  25. Davis BH., et al. “Bethesda International Consensus recommendations on the flow cytometric immunophenotypic analysis of hematolymphoid neoplasia: medical indications”. Cytometry Part B: Clinical Cytometry 72B (2007): S5-S13.
  26. Landgren O., et al. “B cell clones as early markers of chronic lymphocytic leukemia”. The New England Journal of Medicine 360 (2009): 659-667.
  27. Pangalis GA., et al. “B- chronic lymphocytic leukemia, small lymphocytic lymphoma and lymphoplasmacytic lymphoma, including Waldenstorm’s macroglobulinemia: a clinical, morphologic, and biologic spectrum of similar disorders”. Seminar on Hematology 36 (1999): 104-114.
  28. Deneys V., et al. “Atypical lymphocytic leukemia and mantle cell lymphoma immunologically very close: flow cytometric distinction by use of CD20 and CD54 expression”. Leukemia 15 (2001): 1458-1465.
  29. Ballesteros E., et al. “CD5+ low-grade marginal zone B-cell lymphomas with localized presentation”. American Journal of Surgical Pathology 22 (1998): 201-217.
  30. Garcia DP., et al. “Diagnostic Usefulness of CD23 and FMC-7 antigen Expression Patterns in B-cell lymphoma Classification”. American Journal of Clinical Pathology 115 (2001): 258-265.
  31. Fournier S., et al. “CD23 antigen regulation and signaling in chronic lymphocytic leukemia”. Journal of Clinical Investigation 89 (1992): 1312-1321.
  32. kilo MN and Dorfman DM. “The utility of flow cytometric immunophenotypic analysis in the distinction of small lymphocytic lymphoma/ chronic lymphocytic leukemia from mantle cell lymphoma”. American Journal of Clinical Pathology 105 (1996): 451-457.
  33. Ahmad E., et al. “Clinical utility of CD23 and FMC-7 antigen coexistent expression in B-cell lymphoproliferative disorder subclassification”. Cytometry1 (2002): 1-7.
  34. Catovsky D., et al. “Heterogenicity of B-cell leukemias demonstrated by the monoclonal antibody FMC-7”. Blood 58 (1981): 406-408.
  35. Stark AN., et al. “Prolymphocytoid transformation of CLL: a clinical and immunological study of 22 cases”. Leukemia Research 10 (1986): 1225-1232.
  36. Jennings CD and Foon KA. “Recent advances in flow cytometry: application to the diagnosis of hematologic malignancy”. Blood 90 (1997): 2863-2892.
  37. Geisler CH., et al. “Prognostic importance of flow cytometric immunophenotyping of 540 consecutive patients with B-cell chronic lymphocytic leukemia”. Blood 78 (1991): 1795-1802.
  38. Schlette E., et al. “CD79b Expression in chronic lymphocytic leukemia. Association with trisomy 12 and atypical immunophenotype”. Archives of Pathology and Laboratory Medicine 127 (2003): 561-566.
  39. Garcia vela JA., et al. “CD79b expression in B cell chronic lymphocytic leukemia: its implication for minimal residual disease detection”. Leukemia 13 (1999): 1501-1505.
  40. Palumbo GA., et al. “CD200 as a tool for differential diagnosis between B-cell chronic lymphocytic leukemia and mantle cell lymphoma”. Blood 93 (2008): 33.
  41. El-Sewefy DA., et al. “Flow cytometric evaluation of CD200 as a tool for differentiation between chronic lymphocytic leukemia and mantle cell lymphoma”. Egyptian Journal of Haematology2 (2014): 42-46.
  42. Sandes AF., et al. “CD200 Has an Important Role in the Differential Diagnosis of Mature B-Cell Neoplasms by Multiparameter Flow Cytometry”. Cytometry Part B 86B (2014): 98-105.
  43. Alapat D., et al. “Diagnostic usefulness and prognostic impact of CD200 expression in lymphoid malignancies and plasma cell myelomas”. American Journal of Clinical Pathology 137 (2012): 93-100.
  44. Palumbo GA., et al. “CD200 expression may help in differential diagnosis between mantle cell lymphoma and B- cell chronic lymphocytic leukemia”. Leukemia Research 33 (2009): 1212-1216.
  45. Giudice ID., et al. “B-Cell Prolymphocytic Leukemia (B-PLL) and chronic lymphocytic leukemia (CLL) Express Distinct Genomic Profiles”. Blood 106 (2005): 4372.
×

Citation

Citation: Shirin Tarafder. “Immunophenotypic Profile of Patients with Clinically Diagnosed Chronic Lymphocytic Leukemia". Acta Scientific Microbiology 4.1 (2021): 103-113.




Metrics

Acceptance rate30%
Acceptance to publication20-30 days

Indexed In






News and Events


  • Certification for Review
    Acta Scientific certifies the Editors/reviewers for their review done towards the assigned articles of the respective journals.
  • Submission Timeline for Upcoming Issue
    The last date for submission of articles for regular Issues is December 25, 2024.
  • Publication Certificate
    Authors will be issued a "Publication Certificate" as a mark of appreciation for publishing their work.
  • Best Article of the Issue
    The Editors will elect one Best Article after each issue release. The authors of this article will be provided with a certificate of "Best Article of the Issue"

Contact US