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HCV/HBV复合抗原基因的克隆、表达及免疫应答研究

Cloning,Expression and Immune Responses of a Hybrid Antigen Corresponding to Hepatitis C Virus and Hepatitis B Virus

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【作者】 黄建生许谆SaidA.SALEH解咏梅张明徽任大明

【Author】 HUANG Jiansheng,XU Zhun, Said A.SALEH, XIE Yongmei,ZHANG Minghui,REN Daming (National Laboratory of Genetic Engineering, Institute of Genetics,Fudan University,Shanghai 200433)

【机构】 第一军医大学!广州51051复旦大学遗传学研究所遗传工程国家重点实验室!上海200433第二军医大学免疫学教研室!上海200433复旦大学遗传学研究所遗传工程国家重

【摘要】 为探讨HCV/HBV 复合疫苗的可行性,将合成的丙型肝炎病毒(HCV)复合多表位抗原基因PCX与HBsAg 基因连接成PCXS基因,与β-半乳糖苷酶(GZ)基因融合后在大肠杆菌及减毒鼠伤寒沙门氏菌中获得表达.目的蛋白GZ-PCXS可被抗-HBs 及抗-HCV 抗体所特异识别.GZ-PCXS抗原皮下注射免疫ICR小鼠后,诱发了较高水平的抗-GZ-PCXSIgG反应.构建的重组减毒鼠伤寒沙门氏菌SL3261(pWR/PCXS)口服免疫小鼠后,诱发了高水平的CD8+ T细胞增殖反应及抗GZ-PCXSIgG反应.所有免疫小鼠均未见明显的毒副作用.该研究揭示,HCV/HBV 复合抗原可诱发特异性体液免疫及细胞免疫应答,而活菌苗口服可能是理想的免疫途径,为HCV/HBV 双价疫苗研究提供了一定的理论及实验依据.

【Abstract】 In order to explore the possibility of HCV/HBV bivalent vaccine,a synthetic multi epitopes antigen gene PCX of HCV was fused with HBsAg gene.The recombinant gene PCXS was cloned into a β galactosidase expression vector pWR450 1 to highly express a fusion protein GZ PCXS in E.coli JM105 and attenuated Salmonella typhimurium SL3261,whose products were specifically recognized by anti HBs and anti HCV antibodies.After GZ PCXS antigen and recombinant SL3261(pWR/PCXS) bacteria were used to immunize ICR mice,high level of anti GZ PCXS IgG was induced by GZ PCXS antigen.Oral live SL3261(pWR/PCXS) bacteria also elicited significant proliferation responses of CD8 + T cells.All the immunized mice were safe without obvious toxicity.The results showed that the possibility of the bivalent of HCV/HBV vaccines and oral live vaccine might be the best route for this purpose.

【基金】 国家自然科学基金
  • 【文献出处】 中国生物化学与分子生物学报 ,CHINESE JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY , 编辑部邮箱 ,1999年06期
  • 【分类号】R392.1
  • 【被引频次】1
  • 【下载频次】44
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