Flow Cytometry In The Differential Diagnosıs Of Neutropenic Patients
Research Article
DOI:
https://doi.org/10.5281/zenodo.7908431Keywords:
neutropenia, lymphocyte, monocyte, flow cytometry, myelomonocytic antigensAbstract
Introduction: Although immuno-phenotyping with flow cytometry is mainly used in the diagnosis of hematological neoplasia, it also provides important contributions to revealing changes in the immune system. Flow cytometric studies are also frequently performed in neutropenic patients, as changes in neutrophil count may occur in relation to hematological neoplasms and immune system disorders.
Objective: Within the scope of this study, we aimed to elucidate the expressions of lymphocyte, granulocyte, and monocyte subtypes and myelomonocytic antigens on monocytes and neutrophils in neutropenia caused by different reasons and compare them with the results of healthy volunteers.
Method: A total of 24 patients and 13 age-and gender-matched healthy volunteers who applied to our institution have been enrolled in this study. The patient group consisted of individuals with isolated neutropenia. Examination findings, lymph node size, spleen size, and infection findings,and the laboratory parameters, CRP level, and pathological findings in the peripheral smear were recorded. Total blood obtained from the patients has been processed with flow cytometry.
Results: Neutrophil/lymphocyte (N/L) MFI ratio was found to be higher in the patient group than in the control group (p=0.022).A positive correlation was found between leukocyte counts and the percentage of atypical monocyte expressing CD16 (p=0.033). A negative correlation was found between platelet counts and N/L mean fluorescence intensity ratio (p=0.411) and between the platelet count and the percentage of CD11b on neutrophils (p=0.026). The percentage of neutrophils determined on leukocytes and the percentage of natural killer cells (CD30/CD56+) was statistically significantly lower in the patient group (p=0.008, p=0.001 respectively).
Conclusion: Regarding the results of this study one could say that clinically significant changes have been observed in lymphocyte and monocyte subtypes of patients with isolated neutropenia. Granulation was found in the ratio of N/L side scatter, mostly related to neutrophils.
References
Mithoowani S, Cameron L, Crowther MA. Neutropenia. CMAJ. 2022;194(49):E1689. doi:10.1503/cmaj.220499.
Spoor J, Farajifard H, Rezaei N. Congenital neutropenia and primary immunodeficiency diseases. Crit Rev Oncol Hematol. 2019;133:149-162. doi:10.1016/j.critrevonc.2018.10.003
Frater JL. How I investigate neutropenia. Int J Lab Hematol. 2020;42(1):121-132. doi:10.1111/ijlh.13210
Atallah-Yunes SA, Ready A, Newburger PE. Benign ethnic neutropenia. Blood Rev. 2019;37:100586. doi:10.1016/j.blre.2019.06.003
Escrihuela-Vidal F, Laporte J, Albasanz-Puig A, Gudiol C. Update on the management of febrile neutropenia in hematologic patients. Rev Esp Quimioter. 2019;32(2):55-58.
Manohar SM, Shah P, Nair A. Flow cytometry: principles, applications and recent advances. Bioanalysis. 2021;13(3):181-198. doi:10.4155/bio-2020-0267
Robinson JP. Flow cytometry: past and future. Biotechniques. 2022;72(4):159-169. doi:10.2144/btn-2022-0005
Delmonte OM, Fleisher TA. Flow cytometry: Surface markers and beyond. J Allergy Clin Immunol. 2019;143(2):528-537. doi:10.1016/j.jaci.2018.08.011
Steinway SN, LeBlanc F, Loughran TP Jr. The pathogenesis and treatment of large granular lymphocyte leukemia. Blood Rev. 2014;28(3):87-94. doi:10.1016/j.blre.2014.02.001
Lamy T, Loughran TP Jr. How I treat LGL leukemia. Blood. 2011;117(10):2764-74. doi:10.1182/blood-2010-07-296962
Watters RJ, Liu X, Loughran TP Jr. T-cell and natural killer-cell large granular lymphocyte leukemia neoplasias. Leuk Lymphoma. 2011;52(12):2217-25. doi:10.3109/10428194.2011.593276
Zhang J, Xu X, Liu Y. Activation-induced cell death in T cells and autoimmunity. Cell Mol Immunol. 2004;1(3):186-92.
Swerdlow SH, Campo E, Pileri SA, et al. The 2016 revision of the World Health Organization classification of lymphoid neoplasms. Blood. 2016;127(20):2375-90. doi:10.1182/blood-2016-01-643569
Moosic KB, Ananth K, Andrade F, Feith DJ, Darrah E, Loughran TP Jr. Intersection Between Large Granular Lymphocyte Leukemia and Rheumatoid Arthritis. Front Oncol. 2022;12:869205. doi:10.3389/fonc.2022.869205
Lundell R, Hartung L, Hill S, Perkins SL, Bahler DW. T-cell large granular lymphocyte leukemias have multiple phenotypic abnormalities involving pan-T-cell antigens and receptors for MHC molecules. Am J Clin Pathol. 2005;124(6):937-46.
Chandra A, Zhang F, Gilmour KC, et al. Common variable immunodeficiency and natural killer cell lymphopenia caused by Ets-binding site mutation in the IL-2 receptor γ (IL2RG) gene promoter. J Allergy Clin Immunol. 2016;137(3):940-2.e4. doi:10.1016/j.jaci.2015.08.049
Gay D, Kwon O, Zhang Z, et al. Fgf9 from dermal γδ T cells induces hair follicle neogenesis after wounding. Nat Med. 2013;19(7):916-23. doi:10.1038/nm.3181
van Oostveen JW, Breit TM, de Wolf JT, et al. Polyclonal expansion of T-cell receptor-gamma delta+ T lymphocytes associated with neutropenia and thrombocytopenia. Leukemia. 1992;6(5):410-8.
Arber DA, Orazi A, Hasserjian R, et al. The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood. 2016;127(20):2391-405. doi:10.1182/blood-2016-03-643544
Westers TM, Ireland R, Kern W, et al. Standardization of flow cytometry in myelodysplastic syndromes: a report from an international consortium and the European LeukemiaNet Working Group. Leukemia. 2012;26(7):1730-41. doi:10.1038/leu.2012.30
Ogata K. Diagnostic flow cytometry for low-grade myelodysplastic syndromes. Hematol Oncol. 2008;26(4):193-8. doi:10.1002/hon.857
Cherian S, Moore J, Bantly A, et al. Flow-cytometric analysis of peripheral blood neutrophils: a simple, objective, independent and potentially clinically useful assay to facilitate the diagnosis of myelodysplastic syndromes. Am J Hematol. 2005;79(3):243-5. doi:10.1002/ajh.20371
Licona-Limón I, Garay-Canales CA, Muñoz-Paleta O, Ortega E. CD13 mediates phagocytosis in human monocytic cells. J Leukoc Biol. 2015;98(1):85-98. doi:10.1189/jlb.2A0914-458R
Alhan C, Westers TM, Cremers EM, et al. The myelodysplastic syndromes flow cytometric score: a three-parameter prognostic flow cytometric scoring system. Leukemia. 2016;30(3):658-65. doi:10.1038/leu.2015.295.
Boxer L, Dale DC. Neutropenia: causes and consequences. Semin Hematol. 2002;39(2):75-81. doi:10.1053/shem.2002.31911
Boxer LA, Newburger PE. A molecular classification of congenital neutropenia syndromes. Pediatr Blood Cancer. 2007;49(5):609-14. doi:10.1002/pbc.21282
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