Immunophenotypic study of bone marrow erythrokaryocyte differentiation in patients with diffuse large B-cell lymphoma
https://doi.org/10.17650/1726-9784-2018-17-4-52-57
Abstract
Introduction. Previously we»ve shown that there are typical changes in haematopoiesis in diffuse large B-cell lymphoma patients (DLBCL). Attention was paid to the elevation of oxyphylic erythroblasts, as this sign was of prognostic significance. The aim of this work was to see
if there is possible to identify bone marrow erythrokaryocyte maturation stages using flow cytometry method.
Materials and methods. Study was done in 60 DLBCL patients. Elevation of oxyphilic erythrobast levels was noted in 50 patients (83,3 %). Flow cytometric study of bone marrow were done in 30 patients. We used monoclonal antibodies to Glycophorin A, CD45, CD36, CD117, CD71, CD105, CD34, HLA-DR, as well as nuclear staining for SYTO 16, SYTO 41.
Results. Use of Glycophorin A together with of SYTO 16 or SYTO 41 is effective for identification of erythrocaryocytes, main proportion of which is represented by CD71+ and Glycophorin A+ cells, which does not express any precursor markers. In CD45-negative cellular elements there are as well CD71- glycophorin А– cells, which represents mainly precursors – CD34+ CD117+ HLA-DR+/–. Erythro caryocytes with low levels of glycophorin A expression and prominent expression of CD71 (committed to erythroid differentiation) show slight expresiion of CD117 and HLA-DR, but CD34 is completely absent. The informative method for characterization of most mature erythrocaryocyes was the approach based on identification of erythrocaryocytes according to CD36 expression on cels with low SSC (in that case usage of nuclear stain, CD45 and glycophorin A may be omitted as CD36 does not stain erythrocytes). After peak of CD36, CD71, CD105 expression levels of these antigens are declining. Namely that fraction represents most mature erythrocaryocytes. Cor relation of the percentages of that cells with oxyphilic normoblasts (identified morphologically) was near to significant value.
Conclusion. We proved that the most mature erythrocaryocyte fraction may be identified by flow cytometry based on the folowing parameters – SSClow CD71low CD105low CD36+/low.
About the Authors
N. N. Tupitsyn
N.N. Blokhin Russian Cancer Research Center
Russian Federation
24 Kashyrskoe shosse, Moscow 115478
Russian Federation
24 Kashyrskoe shosse, Moscow 115478
Cheng Jiao
N.N. Blokhin Russian Cancer Research Center; Sechenov University
Russian Federation
Russian Federation
24 Kashyrskoe shosse, Moscow 115478
2 build., 8 Trubetskaya St., 119991 Moscow
P. A. Zeynalova
N.N. Blokhin Russian Cancer Research Center; Sechenov University
Russian Federation
Russian Federation
24 Kashyrskoe shosse, Moscow 115478
2 build., 8 Trubetskaya St., 119991 Moscow
References
1. Владимирская Е.Б. Основы кроветворения. В кн.: Гериатрическая гематология. Под ред. Л.Д. Гриншпун, А.В. Пивник. М.: Медиум, 2011. С. 21–44. [Vladimirskaya E.B. Fundamentals of hematopoiesis. In the book: Geriatric hematology. Еd. by L.D. Grinshpun, A.V. Pivnik. M.: Medium, 2011. Pp. 21–44. (In Russ.)].
2. Тупицын Н.Н., Зейналова П.А., Френкель М.А., Османов Е.А. Роль костного мозга в прогнозе диффузной В-крупноклеточной лимфомы. Вопросы биологической, медицинской и фармацевтической химии 2015;1:72–7. [Tupitsyn N.N., Zeynalova P.A., Frenkel M.A., Osmanov E.A. The role of the bone marrow in the prognosis of diffuse B-large cell lymphoma. Voprosy biologicheskoj, medicinskoj i farmacevticheskoj himii = Questions of biological, medical and pharmaceutical chemistry 2015;1:72–7. (In Russ.)].
3. Фалалеева Н.А., Моженкова А.В., Османов Е.А. и др. Особенности эритропоэза больных фолликулярной лимфомой. Вестник ФГБУ «РОНЦ им. Н.Н. Блохина» 2016;3:34–7. [Falaleeva N.A., Mozhenkova A.V., Osmanov E.A. et al. Peculiarities of erythropoiesis in patients with follicular lymphoma. Vestnik FGBU “RONC im. N.N. Blokhin” = Journal of N.N. Blokhin Russian Cancer Research Center 2016;3:34–7. (In Russ.)].
4. Зейналова П.А., Шолохова Е.Н., Френкель М.А. и др. Взаимосвязь иммунофенотипа диффузной В-крупноклеточной лимфомы с особенностями кроветворения у больных. Технологии живых систем 2015;1:63–8. [Zeynalova P.A., Sholokhova E.N., Frenkel M.A. et al. Interrelation of immunophenotype of diffuse B-large cell lymphoma with features of hematopoiesis in patients. Tekhnologii zhivyh sistem = Technologies of living systems 2015;1:63–8. (In Russ.)].
5. Тимонина Е.Г., Френкель М.А., Колбацкая О.П., Подвязников С.О. Характеристика гемопоэза у больных плоскоклеточным раком головы и шеи. Вестник РОНЦ им. Н.Н. Блохина РАМН 2009;20 (3):75–81. [Timonina E.G., Frenkel M.A., Kolbatskaya O.P., Podviaznikov S.O. Characterization of hemopoiesis in patients with squamous cell carcinoma of the head and neck. Vestnik RONC im. N.N. Blokhin RAMN = Journal of N.N. Blokhin Russian Cancer Research Center 2009;20(3):75–81. (In Russ.)].
6. Френкель М.А. Исследование костного мозга в онкологии. Иммунология гемопоэза 2014;12(1–2):18–41. [Frenkel M.A. Study of bone marrow in oncology. Immunologiya gemopoehza = Immunology of hemopoiesis 2014;12(1–2):18–41. (In Russ.)].
7. van Dongen J.J. M., Lhermitte L., Bottcher S. et al. EuroFlow antibody panels for standardized n-dimentional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia 2012;26:1908–75. PMID: 22552007. DOI: 10.1038/leu.2012.120.
8. Stein H., Warnke R.A., Chan W.C. et al. Diffuse large B-cell lymphoma not otherwise specified. In: WHO classification of tumours of haematopoietic and lymphoid tissues (Eds. S.H. Swerdlow, E. Campo, N.L. Harris et al.). Lyon, France: IARC Press, 2008. Pp. 233–237.
9. Cheson B.D., Fisher R.I., Barrington S.F. et al. Recommendation for initial evaluation, staging, and response assessment of Hodgkin and non-Hodgkin lymphoma: The Lugano Classification. J Clin Oncol 2014;32:3059–67. PMID: 25113753. DOI: 10.1200/JCO.2013.54.8800.
10. McGrath K.E., Catherman S.C., Palis J. et al. Delineating stages of erythropoiesis using imaging flow cytometry. Methods 2017;112:68–74. PMID: 27582124. DOI: 10.1016/j.ymeth.2016.08.012.
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For citations:
Tupitsyn N.N., Jiao Ch., Zeynalova P.A. Immunophenotypic study of bone marrow erythrokaryocyte differentiation in patients with diffuse large B-cell lymphoma. Russian Journal of Biotherapy. 2018;17(4):52-57. (In Russ.) https://doi.org/10.17650/1726-9784-2018-17-4-52-57
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