Flowcytometric Cytokine Profiling of T- Lymphocytes in Pediatric Aplastic Anemia
Keywords:
Aplastic Anaemia, Cytokines, Flowcytometry, Immunosuppression, Interferon Gamma
Abstract
Background: Aplastic anemia is a result of complex immune mediated marrow injury. As activated T cells play a pivotal role in immune response, study of cytokine profile in patients of aplastic anaemia (AA) will be of use in understanding the initiation and propagation of the disease. Methods: Thirty children with AA between ages 6 months to 18 years were included along with age & sex matched controls. Complete blood count (CBC) and bone marrow examination was done. They were grouped as Very severe aplastic anemia (VSAA), Severe aplastic anemia (SAA) and Non severe aplastic anemia (NSAA) as per standard guidelines. Intracytoplasmic expression of Type 1 cytokine (IFNγ) and Type 2 cytokine (IL4) was determined by Flow cytometry (FCM). CD8 and CD4 cell counts were also determined by FCM. Result: CD8 cell count was higher than the CD4 counts with reversal of CD4:CD8 ratio in all patients. Mean intracytoplasmic IFNγ expression in CD8 and CD4 cells was higher in cases than in controls (p<0.0001) and showed graded positive correlation with the severity of the disease (p<0.0001). Type 2 cytokine (IL4) did not show any such correlation. Conclusion: IFN gamma expression in T cells can be useful in predicting severity of the disease and also response to immunosuppression.References
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2. Robbins and Cotran. Pathologic basis of disease. Aplastic anaemia 8th edition. Elsevier publishing 2011; 14: 662-665.
3. Nistico A and Young NS. Gamma interferon gene expression in bone marrow of patients with aplastic anaemia .Ann intern med. 1994;120(6):463-469.
4. Mossman TR, Li L, Sad S, et al. Function of CD8 T-cell subsets secreting different cytokine patterns. Semin Immunol.1997; 9: 87–92.
5. Jung T, Schauer U, Heuser C, et al. Detection of intracellular cytokines by flow cytometry. J Immunol Methods. 1993; 159: 197–207.
6. Zoumbos NC, Gascon P, Djeu J, et al. Interferon is a mediator of hematopoietic suppression in aplastic anemia in vitro and possible in vivo. Proc Natl Acad Sci USA 1985;82: 188–192
7. Linenberger ML, Jacobson FW, Bennett LG, Broudy VC, Martin FH, Abkowitz JL. Stem cell factor production by human marrow stromal fibroblasts . Exp hematol 1995;23(10):1104-14
8. Richard A, Davis MH.Henry's Clinical Diagnosis and Management by Laboratory Methods: Laboratory Evaluation of Immunoglobulin function and Humoral immunity .21 st edition 2007;835-875.
9. Feng X, Scheinberg P, Wu CO, et al. Cytokine signature profiles in acquired aplastic anemia and myelodysplastic syndromes.Hematologica 2009;94(12):1743-1747
10. Tsuda H and Yamasaki H. type I and type II T cell profiles in Aplastic Anaemia and refractory anemia. Am J Hematol 2000;64:271-274
11. Dufour C, Corcione A, Svahn J, et al. Interferone gamma and TNF alpha are over expressed in bone marrow T lymphocytes from paediatric patients with aplastic anemia. Br J hematol 2001;115:1023-1031
12. Sloand E, Kim S, Jaroslaw P, et al. Intracellular IFN γ in circulating and marrow T cells detected by flow cytometry and the response to immunosuppressive therapy in patients with aplastic anemia. Blood 2002; 100: 1185-1191.
Published
2020-04-29
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