IL-10对体外培养乙肝患者树突状细胞作用研究.docVIP

IL-10对体外培养乙肝患者树突状细胞作用研究 　　【摘要】目的:研究IL-10对慢性乙型重型肝炎患者树突状细胞的作用。方法:抽取受试慢性乙型重型肝炎患者外周静脉血,分离提取外周血淋巴细胞,分别采用HBcAg、rhIL-4、rhGM-CSF刺激培养获得不同分化的树突状细胞,用不同剂量的IL-10刺激培养树突状细胞,检测各组树突状细胞表型与分泌的细胞因子及体外诱导的淋巴细胞毒活性的变化。结果:IL-10处理的树突状细胞膜具备低表达HLA-DR,CD83,CD86分子,低分泌IFN-γ,IL-12p70的特点,体外诱导减弱自体淋巴细胞毒活性,与DC比较结果具有显著性差异(P 　　【关键词】乙型肝炎;树突状细胞;白细胞介素-10 　　The Effects of Interleukin-10 on Dendritic Cells from Chronic Severe Hepatitis B Patients 　　WANG Sikui SHI Qingfeng YUAN Jianguo SONG Jikui GUAN Shan CHA Yan Wang Weihong Wang Kai Xia Qingjie 　　 　　【Abstract】 Objective:To study the effects of Interleukin-10 on dendritic cells in chronic severe hepatitis B patients.Methods: Monocytes were firstly isolated from fresh peripheral blood of chronic severe hepatitis B patients by Ficoll-Hypaque density gradient centrifugating and then cultured with plastic-adherence method.Dendritic cells were induced and proliferated from the monocytes with granulocyte-macrophage clony stimulating factor and interleukin-4 for 8 days. Dendritic cells cocultured with Hepatitis B Virus core antigen induced mature dendritic cells. Also dendritic cells were exposed to interleukin-10(IL-10).The expression of surface marker on mature/ tolerogenic dendritic cells was detected by FACS.The cytotoxic T lymphocyte activity, as well as the IL-12p70 , IFN-γ secretion were detected. Results: The secretion of IL-12p70,IFN-γ and expression of human leukocyte antigens, as well as the cytotoxic T lymphocyte activity by dendritic cells were inhibited while treated with IL-10. The Hepatitis B Virus core antigen induced dendritic cells maturation, IL-10 cocultured dendritic cells can be maintained in the immature state when Hepatitis B Virus core antigen plused,too. (P[ 1~2],不表达共刺激分子,可以诱导T细胞无能,从而引起自身免疫耐受。Tol DC还可以诱导调节/抑制T细胞来清除免疫反应性T细胞,也可以直接产生IL-10、TGF等细胞因子抑制免疫反应性T细胞,达到诱导外周耐受的目的。IL-10能抑制健康者DC的生长及DC的成熟,产生Tol DC,诱导CTL对抗原产生耐受。我们利用IL-10刺激慢性乙型重型肝炎患者外周静脉血培养的树突状细胞后,检测DC的分化表达和诱导的淋巴细胞毒活性,观察IL-10对慢性乙型重型肝炎患者DC的分化成熟的作用,以期获得稳定的Tol DC,为通过抑制免疫超敏反应治疗慢性乙型重型肝炎的临