【摘要】 目的 探讨HCV/HBV复合抗原PCX/S免疫原性。方法 将已证实具有良好免疫原性的人工合成的HCV复合多表位抗原基因PCX与HBV的S抗原基因融合于真核表达载体pcDNA3.0,构建真核表达载体pcDNA-PCXS。大量提取质粒并免疫小鼠,用ELISA的方法检测抗-HBs和抗-HCVAb;³H-TdR渗入法检测免疫小鼠T淋巴细胞增殖;用⁵¹Cr释放法检测免疫小鼠特异性CTLs杀伤作用。结果 免疫后均能检测到抗-HBsAb和抗-HCVAb,抗体水平随着时间的推移逐渐升高,但后者OD_450nm平均值低于抗-HBsAb的OD_450nm。免疫小鼠诱发针对PCX或S抗原的CTL反应和淋巴细胞转化。结论 复合型PCX/S抗原基因可诱发特异性免疫应答,为HCV/HBV双价疫苗的研究提供一定实验基础。更多还原

【Abstract】 Objective To explore the possibility of the hepatitis C virus / hepatitis B virus bivalent DNA immunization.Methods A synthetic multiple epitope antigen gene PCX of HCV fused with HBsAg gene was cloned into vector pcDNA3.0 to construct eukaryotic expression vectors pcDNA-PCXS. The target genes were sequenced and the plasmids were extracted, then they were used to immunize mice. The anti-HBs and anti-HCV antibodies were detected by ELISA, T lymphocytes proliferation response and the cytotoxicity of splenocytes (CTL) were respectively examined by 3H-TdR and 51Cr-release assays. Results Antibodies against HBsAg and. HCV were detected positively in the sera from immunized mice. But anti-HCV OD450nm values were lower than that of anti-HBsAg. CTL or T cell proliferation response was induced by PCX or S antigen. Conclusion The combined PCX/S antigen gene could induce specific immune response and it might be able to serve as a bivalent DNA vaccines candidate specific for HCV and HBV.更多还原