EVALUATION OF CELL SURFACE ANTIGEN EXPRESSIONS IN CASE OF CHRONIC LYMPHOCYTIC LEUKEMIA

Year 2022, , 183 - 189, 20.04.2022

Filiz Yavaşoğlu , Çiğdem Özdemir , Tülay Köken

https://doi.org/10.18229/kocatepetip.832270

Abstract

OBJECTIVE: Immunophenotyping is critical for the diagnosis of B cell malignancy. In this study, we aimed to evaluate the cell surface antigen expression percentages and densities by flow cytometry method in patients diagnosed with Chronic Lymphocytic Leukemia (CLL) clinically and morphologically.
MATERIAL AND METHODS: Flow cytometry results of peripheral venous blood of 33 CLL patients (12 females / 21 males) were retrospectively analyzed. First, lymphocytes were marked on the CD45-Side Scatter chart and antibody expression percentages and densities in this population were determined.
RESULTS: It was observed that all patients had CD5, CD19, CD20, CD23, CD200 and CD43 antigen expressions as in the classical pattern. Unlike the classical pattern, the expression of FMC7, which is not expected is over 30% in eight patients. While 6 of the cases showed kappa monoclonality, 15 of them showed lambda monoclonality, it was seen that both light chain expressions were below 30% in 11 cases. ZAP 70 expression was found in three cases (16%, 23%, 42%). Looking at the average percentage of antigen expressions of all cases, it is seen that CD5, CD19, CD20, CD23, CD200 and CD43 are above 80%.
CONCLUSIONS: Surface antigen expressions recommended to be checked in the diagnosis of CLL may not follow the rules defined in each case. In these cases, there is a need to search for more specific additional cell markers to enter the diagnostic algorithm.

Keywords

Chronic lymphocytic Leukemia, Flow cytometry, Cell surface antigen

KRONİK LENFOSİTİK LÖSEMİ VAKALARINDA HÜCRE YÜZEY ANTİJEN EKSPRESYONLARININ DEĞERLENDİRİLMESİ

Abstract

AMAÇ: İmmünofenotipleme, B hücre malignitesi teşhisi için kritik öneme sahiptir. Bu çalışmada klinik ve morfolojik olarak Kronik Lenfositik Lösemi (KLL) tanısı almış vakalarımızda akım sitometri yöntemi ile hücre yüzey antijen ekspresyon yüzdelerini ve yoğunluklarını değerlendirmeyi amaçladık.
GEREÇ VE YÖNTEM: 33 KLL hastasının (12 kadın / 21 erkek) periferik venöz kanına ait akım sitometri sonuçları geriye dönük incelenmiştir. İlk olarak CD45-Side Scatter grafiği üzerinde lenfositler işaretlendi ve bu popülasyondaki antikor ekspresyon yüzdeleri ve yoğunlukları belirlendi.
BULGULAR: Tüm hastalarda klasik paternde olduğu gibi CD5, CD19, CD20, CD23, CD200 ve CD43 antijen ekspresyonlarının mevcut olduğu görülmüştür. Klasik paternden farklı olarak ekspresyonu beklenmeyen FMC7’nin sekiz hastada ekspresyonu %30 un üzerindedir. Vakaların 6’sı kappa, 15’i lambda monoklonalitesi gösterirken 11 vakada da her iki hafif zincir ekspresyonunun %30’un altında olduğu görüldü. ZAP 70 ekspresyonu üç vakada (%16, %23, %42) rastlanmıştır. Tüm vakaların antijen ekspresyonlarının ortalama yüzdelerine bakıldığında CD5, CD19, Cd20, Cd23, CD200 ve CD43 ün %80’nin üzerinde olduğu görülmektedir.
SONUÇ: KLL tanısında bakılması önerilen yüzey antijen ekspresyonları her vakada tanımlanan kurallara uymayabilir. Bu vakalarda tanı algoritmasına girecek daha spesifik ek hücre markırlarının araştırılmasına ihtiyaç ortaya çıkmaktadır.

Keywords

Kronik lenfositik lösemi, Akım sitometri, Hücre yüzey antijeni

References

• 1. Wierda WG, Byrd JC, Abramson JS, et al. Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma, Version 4.2020, NCCN Clinical Practice Guidelines in Oncology. J Natl Compr Canc Netw. 2020;18(2):185-217.
• 2. Hallek M, Cheson BD, Catovsky D, 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. 2008;(12):5446-56.
• 3. Eichhorst B, Robak T, Montserrat E, et al. ESMO Guidelines Committee. Chronic lymphocytic leukaemia: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2015;(5)78-84.
• 4. Rawstron AC, Kreuzer KA, Soosapilla A, et al. Reproducible diagnosis of chronic lymphocytic leukemia by flow cytometry: An European Research Initiative on CLL (ERIC) & European Society for Clinical Cell Analysis (ESCCA) Harmonisation project. Cytometry B Clin Cytom. 2018;(1):121-8.
• 5. Matutes E, Owusu-Ankomah K, Morilla R, et al. The immunological profile of B-cell disorders and proposal of a scoring system for the diagnosis of CLL. Leukemia. 1994;8(10):1640-5.
• 6. Wood BL, Arroz M, Barnett D, et al. 2006 Bethesda International Consensus recommendations on the immunophenotypic analysis of hematolymphoid neoplasia by flow cytometry: optimal reagents and reporting for the flow cytometric diagnosis of hematopoietic neoplasia. Cytometry B Clin Cytom. 2007;72(1):14–22.
• 7. Rawstron A, Böttcher S, Letestu R, et al. Improving efficiency and sensitivity: European Research Initiative in CLL (ERIC) update on the international harmonised approach for flow cytometric residual disease monitoring in CLL. Leukemia. 2013;27(1)142–9.
• 8. Almasri NM, Duque RE, Iturraspe J, et al. Reduced expression of CD20 antigen as a characteristic marker for chronic lymphocytic leukemia. Am J Hematol. 1992;40(4)259- 63.
• 9. Ivancević TD, Kurtović NK, Knezević V, et al. The role of immunophenotyping in differential diagnosis of chronic lymphocytic leukemia. Srp Arh Celok Lek. 2014;142(3-4):197- 203.
• 10. Fang C, Zhuang Y, Wang L, et al. High levels of CD20 expression predict good prognosis in chronic lymphocytic leukemia. Cancer Sci. 2013;104(8):996-1001.
• 11. Bikah G, Lynd FM, Aruffo AA, et al. A role for CD5 in cognate interactions between T cells and B cells, and identification of a novel ligand for CD5. Int Immunol 1998;10(8):1185- 96.
• 12. Lee JH, Noh J, Noh G, et al. IL-10 is predominantly produced by CD19(low)CD5(+) regulatory B cell subpopulation: characterisation of CD19 (high) and CD19(low) subpopulations of CD5(+) B cells. Yonsei Med J. 2011;52(5):851-5.
• 13. Cartron G, Linassier C, Bremond JL, et al. CD5 negative B-cell chronic lymphocytic leukemia: clinical and biological features of 42 cases. Leuk Lymphoma. 1998;31(1-2):209-16.
• 14. Demir C, Kara E, Ekinci Ö, et al. Clinical and Laboratory Features of CD5-Negative Chronic Lymphocytic Leukemia. Med Sci Monit. 2017;23:2137-42.
• 15. Deneys V, Michaux L, Leveugle P, et al. Atypical lymphocytic leukemia and mantle cell lymphoma immunologically very close: flow cytometric distinction by the use of CD20 and CD54 expression. Leukemia. 2001;15(9):1458-65.
• 16. Sarfati M. CD23 and chronic lymphocytic leukemia. Blood Cells.1993;19(3):591-6.
• 17. Kilo MN, Dorfman DM. The utility of flow cytometric immunophenotypic analysis in the distinction of small lymphocytic lymphoma/chronic lymphocytic leukemia from mantle cell lymphoma. Am J Clin Pathol. 1996;105(4):451-7.
• 18. Fateh, Numan. Mantle Cell Lymphoma Misdiagnosed as Chronic Lymphocytic Leukemia: Optimization of Diagnostic Approach. Journal of Clinical & Experimental Oncology. 2017;(6)1-3.
• 19. DiRaimondo F, Albitar M, Huh Y, et al. The clinical and diagnostic relevance of CD23 expression in the chronic lymphoproliferative disease. Cancer. 2002;94(6):1721-30.
• 20. Dadmarz R, Cawley JC. Heterogeneity of CLL: high CD23 antigen and alpha IFN receptor expression are features of favourable disease and of cell activation. Br J Haematol. 1988;68(3):279-82.
• 21. Lavabre-Bertrand T, Exbrayat C, et al. CD23 antigen density is related to serum gamma globulin level, bone marrow reticulin pattern, and treatment in B chronic lymphocytic leukemia. Leuk Lymphoma. 1994;13(1-2):89-94.
• 22. Wright GJ, Jones M, Puklavec MJ, et al. The unusual distribution of the neuronal/lymphoid cell surface CD200 (OX2) glycoprotein is conserved in humans. Immunology. 2001;102(2):173-9.
• 23. Moreaux J, Hose D, Reme T, et al. CD200 is a new prognostic factor in multiple myeloma. Blood. 2006;108(13):4194-7.
• 24. Damiani D, Tiribelli M, Raspadori D, et al. Clinical impact of CD200 expression in patients with acute myeloid leukemia and correlation with other molecular prognostic factors. Oncotarget. 2015;6(30):30212-221.
• 25. Palumbo GA, Parrinello N, Fargione G, et al. CD200 expression may help in differential diagnosis between mantle cell lymphoma and B-cell chronic lymphocytic leukemia. Leuk Res. 2009;33(9):1212-16.
• 26. Sandes AF, de Lourdes Chauffaille M, Oliveira CR, et al. CD200 has an important role in the differential diagnosis of mature B-cell neoplasms by multiparameter flow cytometry. Cytometry B Clin Cytom. 2014;86(2):98-105.
• 27. Delgado J, Matutes E, Morilla AM, et al. Diagnostic significance of CD20 and FMC7 expression in B-cell disorders. Am J Clin Pathol. 2003;120(5):754-9.
• 28. Reddy P, Dabbas B, Gama M, et al. FMC-7 Expression Identifies Phenotypically Atypical Chronic Lymphocytic Leukemia with Distinct Clinical and Molecular Genetic Features. Blood. 2012;120 (21): 2478.
• 29. Criel A, Wlodarska I, Meeus P, et al. Trisomy 12 is uncommon in typical chronic lymphocytic leukaemias. Br J Haematol. 1994;87:523-8.
• 30. Peltomäki P, Bianchi NO, Knuutila S, et al. Immunoglobulin kappa and lambda light chain dual genotype rearrangement in a patient with kappa-secreting B-CLL. Eur J Cancer Clin Oncol. 1988;24(7):1233-8.
• 31. Bhola RK, Das PK, Pradhan S, et al. Multiplexing 8 colors with 12 antibodies in a single lymphoid screening tube by flow cytometry for evaluating suspected chronic lymphoproliferative disorders (CLPD). J Hematopathol. 2020;13:13–24.
• 32. Alves G. VA, Silva L. KF, Albuquerque D. GB, et al. Use of Flow Cytometry to Evaluate the Immunophenotypic Profile of Patients with Chronic Lymphocytic Leukemia from Rio Grande Do Norte State, Brazil. Blood 2017;130(1):5325.
• 33. Paiva A.S, Jardim A.S, Soares V.L, et al. Diagnostic Value of Kappa/Lambda Ratios Determined By Flow Cytometric Analysis of Peripheral Blood and Bone Marrow Specimens in B-Cell Chronic Proliferative Disease. Blood. 2018;132:5431.
• 34. Abdel-Ghafar AA, El Din El Telbany MA, Mahmoud HM, El-Sakhawy YN. Immunophenotyping of chronic B-cell neoplasms: flow cytometry versus immunohistochemistry. Hematol Rep. 2012;4(1):e3.
• 35. Zhou J, Tewari A, Nassiri M, et al. Changes in Antigen Expression in a Follow-up of Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma. J Leuk. 2015;3:177.
• 36. Hamblin TJ, Davis Z, Gardiner A, et al. Unmutated Ig VH Genes Are Associated With a More Aggressive Form of Chronic Lymphocytic Leukemia. Blood. 1999;94 (6):1848–54.
• 37. Wiestner A, Rosenwald A, Barry TS, et al. ZAP-70 expression identifies a chronic lymphocytic leukemia subtype with unmutated immunoglobulin genes, inferior clinical outcome, and distinct gene expression profile. Blood 2003;101:4944–51.
• 38. Crespo M, Bosch F, Villamor N, et al. ZAP-70 expression as a surrogate for immunoglobulin-variable-region mutations in chronic lymphocytic leukemia. N Engl J Med. 2003;348:1764–75.
• 39. Liu Y, Wang Y, Yang J, Bi Y, Wang H. ZAP-70 in chronic lymphocytic leukemia: A meta-analysis. Clin Chim Acta. 2018;483:82-8.