CD8^+T淋巴细胞非溶细胞功能抑制HepG2.2.15细胞表达HBV的实验研究  

Effect of CD8^+T Cell on HBV Replication of HepG2.2.15 Cells by Noncytolytic Functions

作　　者：唐祖明[1] 张胜子[1] 常静霞[2] 郑纪山[3] 

机构地区：[1]东南大学生物电子学国家重点实验室,江苏南京210096 [2]解放军第八一医院,江苏南京210002 [3]南京军区军事医学研究所,江苏南京210002

出　　处：《临床医学工程》2010年第2期1-3,共3页Clinical Medicine & Engineering

基　　金：国家自然科学基金资助项目(编号:30671929)

摘　　要：目的观察慢性乙肝患者病毒特异性CD8+T细胞体外非溶细胞功能抑制HepG2.2.15细胞表达乙型肝炎病毒的作用。方法选择低溶细胞活性的HBcAg肽特异性CD8+T细胞克隆(效应细胞)与HepG2.2.15细胞(靶细胞)以1∶10共同培养,于24h、48h和72h收集培养上清,通过检测其中细胞因子及HBV产物的变化,观察CD8+T克隆对HBV的抑制作用。用抗体中和法观察CD8+T细胞分泌的细胞因子被封闭后HBV抑制的变化。结果HBV特异性CD8+T克隆与靶细胞共育后,培养上清可检出高水平IFN-γ和少量TNF-α。共育后对HBsAg、HBeAg和HBV-DNA的最高抑制率分别为71.2%、68.5%和78.3%,均在72h。IFN-γ和TNF-α单独和同时被抗体封闭后,对HBV-DNA的抑制率显著下降。对靶细胞的最大细胞毒活性是7.2%(24h)。结论IFN-γ和TNF-α是CD8+T细胞非溶细胞机制清除病毒的主要效应分子。Objective To investigate the effects ofHBV-specific CD8^＋T cells on inhibiting HBV replication in vitro. Methods By using coculture of HBcAg-specific CD8^＋T cell clone （effector cells）that were characterized by lower cytotoxicity, with HepG2.2.15 cell （target cells） at E：T ratios of 1 ： 10, and monitoring HBV （HBsAg, HBeAg, and HBV-DNA）in coculture supernatants at 24 h, 48 h and 72 h, the percentage of decrease in HBV replication level were observed. Furthermore, blocking experiment with neutralizing mAbs to cytokine was performed to evaluate the effect of the cytokine. Results CD8^＋F clone produced high levels of IFN-γ and lower levels of TNF-α following coculture with HepG2.2.15 cells. FIBsAg, HBeAg and HBV-DNA in coculture supematants were significantly reduced, and the greatest effect were observed at 72 h by 71.2%、 68.5% and 78.3%, respectively. The reduction of HBV-DNA decreased followed by using neutralizing mAbs to IFN-γ and TNF-α. The maximum activity of cytotoxicity of target cells at 24 h by 7.2%. Conclusion CD8^＋T cell control of HBV replication by noncytolytic mechanisms is mainly mediated by IFN-3, and TNF-α.

关 键 词：乙型肝炎病毒 CD8+T细胞 HEPG2.2.15细胞 干扰素-Γ 肿瘤坏死因子-α 

分 类 号：R512.6[医药卫生—内科学]