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为研发安全广谱有效的丙型肝炎病毒(Hepatitis C virus,HCV)T细胞疫苗,本研究构建了表达HCV截短型非结构蛋白3(Nonstructural protein 3,NS3)与核心蛋白(core)融合抗原的重组腺病毒疫苗。体外免疫荧光及蛋白印迹实验表明该融合抗原可有效表达;小鼠免疫结果表明该重组腺病毒疫苗除了激发抗原特异的抗体反应外,还可激发较强的针对NS3抗原特异的T细胞免疫应答。该T细胞免疫应答主要表现为IFN-γ+与TNF-α+CD4+T细胞亚群。采用异型(JFH1,2a型)HCV重组痘病毒接种小鼠进行保护效果分析,与对照组相比,表达截短型NS3与core融合抗原的重组腺病毒疫苗2针免疫后可产生明显的交叉免疫保护。本研究为进一步研究HCV免疫保护机制及新型疫苗研制提供了参考。
In order to develop a safe and broad-spectrum Hepatitis C virus (HCV) T cell vaccine, a recombinant plasmid expressing HCV truncated Nonstructural protein 3 (NS3) and core fusion antigen Recombinant adenovirus vaccine. In vitro immunofluorescence and Western blotting showed that the fusion antigen was expressed efficiently. The results of immunization showed that the recombinant adenovirus vaccine could stimulate the T-cell immune response specific to NS3 antigen in addition to the antigen-specific antibody response. The T cell immune response mainly manifested as IFN-γ + and TNF-α + CD4 + T cell subsets. The protective effect of mice vaccinated with heterologous (JFH1,2a) HCV recombinant poxvirus was analyzed. Compared with the control group, 2-needle immunized recombinant adenovirus vaccine expressing truncated NS3 and core fusion antigen could produce significant cross-immunization protection. This study provides a reference for further study of HCV immune protection mechanism and new vaccine development.