The Functional Study Of Immune Cell Subsets In Patients With Immune Thrombocytopenia

Objective: To explore Ig G concentration in B cells supernatants and the concentrations of cytokines in Dendritic cells, CD8+T cells and CD4+T cells supernatants, we can investigate the relationship between the function of immune cell subsets in patients with Immune Thrombocytopenia and disease occurrence; moreover, further study on the pathological mechanism will be helpful for the diagnosis and treatment of ITP.Methods:1.The peripheral blood mononuclear cells(PBMCs) of ITP patients and health adults were separated by Ficoll-Hypaque method; 2.The B cells, Dendritic cells, CD8+T cells and CD4+T cells were isolated by the Magnetic cell sorting(MACS) from from the PBMCs respectively; 3.The purity of B cells, Dendritic cells, CD8+T cells and CD4+T cells were detected by flow cytometry respectively; 4. Enzyme-linked immunosorbent assay(ELISA) was employed to determine PAIg G concentration in B cell supernatants; The concentrations of IL-2, IL-6, IL-12, IL-27, TNF-? and IFN-? in Dendritic cells supernatants were determined by ELISA; The concentrations of IL-2, IL-12, IL-17, TNF-?, IFN-? and TGF-? in CD8+T cells supernatants were determined by ELISA; The concentrations of IL-2, IL-4, IL-10, IL-17, TNF-?, IFN-? and TGF-? in CD4+T cells supernatants were determined by ELISA;Results: 1.The Ig G concentration in B cell supernatants was(16.79±1.12 pg /ml) in ITP group, which was significantly higher than that(5.18±0.79pg/ml) in normal control group(p<0.01); 2.The expression level of cytokines IL-12, IL-27 in Dendritic cells supernatants were(51.39±4.55pg/ml, 178.07±50.31pg/ml) in ITP group, which was significantly higher than that(23.64±2.82pg/ml, 92.74±43.57 pg/ml) in normal control group(p<0.01); 3.The expression level of cytokine TGF-? in CD8+T cells supernatants was(203.26±48.51pg/ml) in ITP group, which was significantly lower than that(395.80±74.05pg/ml) in normal control group(p<0.05); 4.The expression level of cytokines IL-4, IL-10, IL-17 in CD4+T cells supernatants were(17.26±2.11pg/ml, 4.12±0.39pg/ml, 15.89±1.31pg/ml) in ITP group, which was significantly higher than that(10.53±1.32pg/ml, 2.10±0.44pg/ml, 9.42±0.24pg/ml) in normal control group(p<0.01), While the expression level of cytokine TGF-? was(379.47±44.83pg/ml) in ITP group, which was significantly lower than that(667.42±67.25pg/ml) in normal control group(p<0.01);Conclusion:1.The PAIg G concentration in ITP group was significantly higher than that in normal control group, and suggest the detection may have a relative significance in clinical diagnosis.2. The expression level of cytokines IL-12, IL-27 in Dendritic cells supernatants in ITP group was significantly higher than that in normal control group,and Dendritic cells play an important role in ITP pathogenesis through secreting IL-12 and IL-27. 3.CD8+T cell may play an important role in ITP pathogenesis through secreting TGF-?. 4.The abnormality of Th cells in patients with ITP implied that ITP was a kind of autoimmune disease with Th2 predominant pathogenic features. 5. IL-17 and TGF-? were involved in the pathogenesis of ITP, that may exist Th17/Tregs balance disorders in patients with ITP.