| HBcAg肽特异性CD8+T细胞抑制乙型肝炎病毒的体外实验研究 |
| 投稿时间：2010-01-11 修订日期：2010-03-12 点此下载全文 |
| 引用本文：郑纪山,唐雨德,何长伦,高健,常静霞.HBcAg肽特异性CD8+T细胞抑制乙型肝炎病毒的体外实验研究[J].医学研究杂志,2010,39(5):24-27 |
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| 基金项目:国家自然科学基金资助项目（30671929） |
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| 中文摘要:目的观察HBV感染者的HBcAg肽特异性CD8+T细胞体外抑制乙型肝炎病毒（HBV）的作用和探讨非溶细胞机制清除病毒的效应分子。方法以HepG2.2.15细胞为靶细胞。用HLA-A匹配的HBcAg肽特异性CD8+T细胞克隆（效应细胞）与靶细胞（效靶比例1∶50）共同培养，于24h、48h和72h收集培养上清，通过检测其中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IFN-γ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。共育后对HBsAg、 HBeAg和HBV-DNA的最高抑制率分别为54.55%、5036%和74.55%，均在72h。IFN-γ被抗体封闭后，对HBV DNA的抑制率显著下降，24h和48h分别为6.22%和17.48%。细胞毒活性最高见于24h，15.66%。结论①病毒特异性CD8+T细胞对靶细胞中HBV的清除既有溶细胞机制，也有非溶细胞机制参与；②IFN-γ是非溶细胞机制清除病毒的主要效应分子。 |
| 中文关键词:乙型肝炎病毒 特异性CD8+T细胞 干扰素-γ HepG2.2.15细胞 |
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| HBcAg-specific CD8+T Cells Inhibit HBV Replication in vitro |
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| Abstract:ObjectiveTo investigate the effects of HBcAg-specific CD8+T cells on inhibiting HBV replication in vitro,and to search the cytokine of noncytolytic mechanisms in viral clearance. MethodsBy the method of coculture of HepG2.2.15 cell (target cells) with HLA-A2 matched HBcAg-specific CD8+T cell clone (effector cells) at E:T ratios of 1∶50, and monitoring HBV production (HBsAg,HBeAg, and HBV-DNA)in coculture supernatants at 24h,48h and 72h, the percentage of decrease in HBV replication level was observed. Furthermore, blocking experiment with neutralizing mAbs to IFN-γ was performed to evaluate the effect of this cytokine. ResultsCD8+T clone produced high levels of IFN-γfollowing coculture with 2.2.15 cells. HBsAg,HBeAg and HBV-DNA in coculture supernatants were significantly reduced, and the greatest effect was observed at 72h by 54.55%,50.36% and 74.55%, respectively. The reduction of HBV DNA was decreased followed by using neutralizing mAbs to IFN-γ. The maximum activity of cytotoxicity of target cells was at 24h by 15.66%. Conclusion①HBV- specific CD8+T cells inhibit HBV replication by cytolytic and noncytolytic mechanisms.②The effect of noncytolytic mechanisms is mainly mediated by IFN-γ. |
| keywords:Hepatitis B virus Specific CD8+T cells IFN-γ HepG2.2.15 cells |
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