目的本研究旨在构建一种包含丙型肝炎病毒(HCV)保守区基因的新型DNA疫苗,并在小鼠模型中使用电转技术优化其免疫原性。方法首先,我们构建了包含HCV非结构蛋白NS3和核心蛋白Core部分基因序列的DNA疫苗,并证实了其表达;然后采用不同的体内电转方式于第0、4周分别免疫BALB/c小鼠,比较分析不同免疫方案的体液免疫(特异性IgG与抗体亚类)与细胞免疫应答(IFN-γ ELISPOT)的效果。结果使用电转技术可显著增强新型DNA疫苗免疫原性,采用皮内注射加卡钳电极电转的方式产生最强NS3特异性T细胞免疫反应。结论包含HCV保守区基因的新型DNA疫苗可通过优化电转技术增强免疫应答效果。这为我们下一步优化HCVDNA疫苗的免疫方案提供了依据。
Objective To characterize the immunogenicity in gene immunization of the conserved regions of hepatitis C virus(HCV) based on different delivery strategies. Methods We first constructed a novel DNA vaccine encoding a fusion gene( from partial NS3 and Core) of HCV. Then we compared different protocols based on naked DNA injection twice or DNA injection with gene electrotransfer(GET) in BALB/c mice. The immune response was measured by antibody ELISA and by IFN-gamma ELISPOT. Results Our data showed that a protocol based on intradermally injection of DNA with optimal GET induced the strongest humoral and cellular immunity, and DNA with GET induced a substantially higher anti-NS3/Core T cell response than naked DNA injection. Conclusion Our data suggest that DNA vaccines encoding NS3/Core fusion protein of HCV immunized by the present strategy could merit further study in the context of future prophylactic and therapeutic HCV T cell based vaccines.