PTD-HBcAg融合蛋白诱导特异性CTL在HepG2.2.15细胞抑制HBV复制的研究  

Study on specific cytotoxic T lymphocytes induced by PTD-HBcAg fusion protein to inhibit HBV replication in HepG2215 cells

作　　者：赖静兰[1] 陈小华[1] 潘庆春[1] 臧国庆[1] 

机构地区：[1]上海交通大学附属第六人民医院感染病科,上海市200233

出　　处：《实用肝脏病杂志》2010年第3期166-169,共4页Journal of Practical Hepatology

基　　金：国家自然科学基金资助项目(30571669)

摘　　要：目的探讨PTD-HBcAg融合蛋白诱导小鼠体内特异性CTL并抑制HepG2.2.15细胞HBV复制的作用。方法 PTD-HBcAg、HBcAg和阴性对照分别与等体积的弗氏佐剂乳化后皮下免疫小鼠;第14d,分离脾淋巴细胞并分别用PTD-HBcAg、HBcAg、PTD和PBS加强刺激后收集上清,检测细胞因子IFN-γ、IL-12、IL-4和IL-10;刺激后的淋巴细胞作为效应细胞与HepG2.2.15细胞共培养,检测,效应细胞对HBsAg、HBVDNA的抑制作用及对HepG2.2.15细胞、HepG2细胞的杀伤效果。结果 PTD-HBcAg组分泌的IFN-γ、IL-12、IL-4和IL-10与HBcAg组和阴性对照组比较差异有统计学意义(P<0.05);PTD-HBcAg融合蛋白组较HBcAg组和阴性对照组有更明显的病毒抑制作用(P<0.05);PTD-HBcAg组对HepG2.2.15细胞的杀伤率明显高于HBcAg组和阴性对照组(P<0.05)。结论 PTD-HBcAg可诱导HBV特异性CTL,能有效抑制HepG2.2.15细胞HBV的复制。Objective To study the effects of PTD-HBcAg inducing HBV specific cytotoxic T lymphocytes in mice on inhibiting HBV replication in HepG2.2.15 cells. Methods BALB/c mice were subcutaneously immunized at the tail base with 100μl of freshly prepared emulsion containing with PTD-HBcAg,HBcAg,PTD,PBS and an equal volume of CFA. 14days later,T lymphocytes in spleen were isolated and stimulated with corresponding proteins to detect the level of IFN-Υ,IL-12,IL-4 and IL-10 in the supernatants. The stimulated T lymphocytes as effective cells were cocultured with HepG2.2.15 cells to determine the levels of HBsAg and HBV DNA and to observe the cytotoxicity of effective cells to HepG2.2.15 cells. Results The cytokines in PTD-HBcAg groups were markedly higher than HBcAg groups and normal controlled groups;In comparison with normal controlled groupsand HBcAg groups,HBsAg and HBV DNA decreased more obviously in PTD-HBcAg groups;The T lymphocytes stimulated by PTD-HBcAg could kill HepG2.2.15 cells more effectively than others. Conclusions PTD-HBcAg can highly promote HBV specific CTL to inhibit HBV replication in HepG2.2.15 cells.

关 键 词：乙型肝炎病毒 HEPG2.2.15细胞 PTD-HBcAg 融合蛋白 细胞毒性T淋巴细胞 

分 类 号：R512.62[医药卫生—内科学]