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LETTER TO EDITOR  
Year : 2020  |  Volume : 63  |  Issue : 3  |  Page : 505-506
Levels of expression of CD45 for normal lymphocytes in different leukemic cases by flowcytometry


Laboratory Oncology Unit, Dr. B.R.A Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India

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Date of Submission09-Mar-2019
Date of Acceptance20-Mar-2019
Date of Web Publication7-Aug-2020
 

How to cite this article:
Rai S, Singh S, Gupta R. Levels of expression of CD45 for normal lymphocytes in different leukemic cases by flowcytometry. Indian J Pathol Microbiol 2020;63:505-6

How to cite this URL:
Rai S, Singh S, Gupta R. Levels of expression of CD45 for normal lymphocytes in different leukemic cases by flowcytometry. Indian J Pathol Microbiol [serial online] 2020 [cited 2020 Oct 30];63:505-6. Available from: https://www.ijpmonline.org/text.asp?2020/63/3/505/291659




Dear Editor,

A lymphocyte is one of the subtypes of white blood cell in a vertebrate's immune system. Lymphocytes include natural killer cells which function in cell-mediated, cytotoxic innate immunity, T cells for cell-mediated, cytotoxic adaptive immunity and B cells for humoral, antibody-driven adaptive immunity.[1] The common leukocyte antigen, CD45, is a complex family of high molecular weight glycoproteins expressed on all lymphohematopoietic cells. CD45 exhibits protein tyrosine phosphatase activity and plays a role in the regulation of cell differentiation. It is present on normal and malignant cells of the myeloid, T- and B-cell lineage. On B-cell lymphocytes, CD45 expression increases during normal cell maturation in bone marrow and remains stable on mature cells. It is widely known that inpatients with B-acute lymphocytic leukemia (B-ALL), CD45 expression is decreased.[2],[3],[4],[5] In contrast, only scarce and contradictory information is available on CD45 expression in patients with chronic B-cell malignancies. For example, Caldwell and Patterson [1] found decreased CD45 expression on chronic lymphocytic leukemia (CLL) cells, whereas Lavabre-Bertrand et al.[3] reported that CD45 expression on CLL cells was variable but comparable to normal B lymphocytes. For patients with hairy cell leukemia (HCL), decreased as well as increased CD45 expression has been described when compared with normal B lymphocytes.[1],[4] In the present study, we performed a quantitative analysis of levels of expression of CD45 for normal lymphocytes in various leukemic malignancies (B-ALL, T-ALL, AML and CLL). Normal blood samples were obtained from 10 healthy volunteers. Flow cytometric immunophenotyping records of 146 cases including B-ALL, T-ALL, AML and CLL retrospectively evaluated. Standard method for cells staining was used in all the cases. CD45 bright positive lymphocytes were gated on CD45 vs SSC plot to exclude normal immature B-cell progenitors.

Data were analyzed using Kaluza software v2.0 (Beckman Coulter, USA). Data were recorded as the mean fluorescence intensity (MFI) for signal and noise. Signal-to–noise (S/N) ratio was calculated for CD45 expressed lymphocytes. In healthy adults, the MFI and S/N ratio of CD45 expression for the whole lymphocyte population (n = 14) was 82.2 and 62.24, respectively. Within the normal lymphocyte population, the S/N ratio for CD45 expression was variable in different leukemic states with progressively increasing values from B-ALL (254.4; n = 42), T-ALL (280.2; n = 42), AML excluding APL (335.4; n = 42) and APL (455.5; n = 20). Finally, we evaluated CD45 expression for the whole lymphocyte population in APL patients (n = 20) was more than other cases [Figure 1] and [Table 1].
Figure 1: S/N ratio for different leukemic cases with their expression in bar graph

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Table 1: The MFI and S/N ratio for different leukemic cases and their signific

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In the present report, we described that, when compared with CD45 expression on the normal lymphocyte population of leukemic malignancies, B-ALL cells have decreased CD45 expression than APL, AML and T-ALL. When compared with CD45 expression on the whole lymphocyte population of healthy adults, CD45 expression was lower than other malignancies. Therefore, above result explains that normal lymphocytes expression was not stable in all the malignancies as compared with healthy sample. The expression of CD45 and hence SNR is variable in different leukemic states. This information may be useful in deciding on appropriate fluorcochrome for CD45 while designing panels for leukemia immunophenotyping in clinical practice.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

1.
Berrington JE, Barge D, Fenton AC, Cant AJ, Spickett GP. Lymphocyte subsets in term and significantly preterm UK infants in the first year of life analysed by single platform flow cytometry. Clin Exp Immunol 2005;140:289-92.  Back to cited text no. 1
    
2.
Caldwell CW, Patterson WP. Relationship between CD45 antigen expression and putative stages of differentiation in B-cell malignancies. Am J Hematol 1991;36:111-5.  Back to cited text no. 2
    
3.
Lavabre-Bertrand T, Duperray C, Brunet C, Poncelet P, Exbrayat C, Bourquard P, et al. Quantification of CD24 and CD45 antigens in parallel allows a precise determination of B-cell maturation stages: Relevance for the study of B-cell neoplasias. Leukemia 1994;8:402-8.  Back to cited text no. 3
    
4.
Caldwell CW, Patterson WP. Relationship between T200 antigen expression and stages of B-cell differentiation in resurgent hyperplasia of bone marrow. Blood 1987;70:1165-72.  Back to cited text no. 4
    
5.
Shah VO, Civin CI, Loken MR. Flow cytometric analysis of human bone marrow IV. Differential quantitative expression of T-200 common leukocyte antigen during normal hemopoiesis. J Immunol 1988;140:1861-7.  Back to cited text no. 5
    

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Correspondence Address:
Sandeep Rai
C-801, Fortune Residency, Rajnagar Extension, Ghaziabad - 201 017, Uttar Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/IJPM.IJPM_188_19

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