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

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Citation

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

Copyright

Copyright: © 2021 Shirin Tarafder. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.