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目的探讨慢性乙型肝炎患者树突状细胞(dendriticcells,DCs)与CD4+Th细胞亚群分化的关系。方法分离慢性乙型肝炎患者外周血单核细胞,以rhIL-4(50ng/mL)、rhGM-CSF(10ng/mL)和rhTNF-α(100μ/mL)诱导培养DC。以流式细胞仪检测DCs表面CD1a、CD83、CD80、CD和HLA-DR分子的表达情况。免疫磁珠分离外周血CD4+T细胞亚群,PMA+Ionomycin刺激后胞内荧光染色,流式细胞仪检测Th细胞内特征性细胞因子IFN-γ/IL-4以判断Th1/Th2分化。ELISA法检测DCs或Th细胞培养上清中IL-6、IL-12、IFN-γ和IL-4的含量。结果慢性乙型肝炎患者的DCs表达CD1a、CD83、CD80、CD86和HLA-DR分子的水平明显低于正常人;培养至第7天,慢性乙型肝炎患者DCs分泌的IL-12水平低于正常人,而分泌的IL-6水平高于正常人。与正常人相比,慢性乙型肝炎患者外周血中Th1细胞占CD4+T细胞的百分比较低,Th细胞培养上清中IFN-γ的量也较低。患者DCs与同种异体的健康人Th细胞共培养,刺激Th1型细胞因子IFN-γ产生的能力低于正常人。结论慢性乙肝患者体内DCs功能的异常可能导致了外周血Th1分化不足。
Objective To investigate the relationship between dendritic cells (DCs) and the differentiation of CD4 + Th cells in patients with chronic hepatitis B. Methods Peripheral blood mononuclear cells from patients with chronic hepatitis B were isolated and cultured DCs were induced by rhIL-4 (50ng / mL), rhGM-CSF (10ng / mL) and rhTNF-α (100μ / mL). Flow cytometry was used to detect the expression of CD1a, CD83, CD80, CD and HLA-DR on the surface of DCs. Immunomagnetic separation of peripheral blood CD4 + T cell subsets, PMA + Ionomycin stimulated by intracellular fluorescence staining, flow cytometry Th cells characteristic cytokines IFN-γ / IL-4 to determine Th1 / Th2 differentiation. The contents of IL-6, IL-12, IFN-γ and IL-4 in DCs or Th cell culture supernatants were detected by ELISA. Results The levels of CD1a, CD83, CD80, CD86 and HLA-DR in DCs from patients with chronic hepatitis B were significantly lower than those from normal controls. Secretion of IL-12 by DCs from patients with chronic hepatitis B was lower than normal Human, while secreting IL-6 levels higher than normal. Compared with normal people, the percentage of Th1 cells in CD4 + T cells in peripheral blood of patients with chronic hepatitis B is lower, and the amount of IFN-γ in the culture supernatant of Th cells is also lower. DCs were co-cultured with allogeneic healthy human Th cells and their ability to stimulate Th1-type cytokine IFN-γ production was lower than that of normal individuals. Conclusion The abnormal function of DCs in patients with chronic hepatitis B may lead to the lack of Th1 differentiation in peripheral blood.