IL-10对体外培养的乙肝患者的树突状细胞的作用研究(1)
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【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<0.05)。IL-10共培养的树突状细胞可在HBcAg刺激下维持不成熟状态;而普通培养的DC在HBcAg刺激下迅速成熟,高表达HLA-DR,CD83,CD86分子,分泌大量IFN-γ,IL-12p70,再加入IL-10不能逆转。结论:IL-10可抑制慢性乙型重型肝炎患者DC的成熟,并能诱导自体淋巴细胞毒活性减弱。
【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<0.05).Conclusion: IL-10 can inhibit dendritic cells maturation, and produced tolerogenic dendritic cells can reduced cytotoxicity of autologous lymphocytes.
【Key words】hepatitis B;dendritic cells;interleukin-10
【中图分类号】R620【文献标识码】A【文章编号】1005-0515(2010)011-0009-02
慢性乙型重型肝炎是由乙型肝炎病毒(HBV)感染引起的死亡率较高的疾病,其发病机理被认为主要是由于HBV感染引发的免疫超敏反应。树突状细胞(Dendrtic cell, DC)是已知功能最强的专职抗原呈递细胞,在免疫应答中居于中心调控地位。未成熟DC,称为致耐受DC (tolerogenic DC,Tol DC) [ 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 ......
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