表达丙型肝炎病毒NS3和Core融合蛋白基因的DNA疫苗免疫方案优化  

Enhancement of the immune response of a novel DNA vaccine encoding conserved NS3 and Core fusion gene of HCV injected by intradermal electrotransfer in mice

作　　者：殷霄[1] 鲁健[2] 谭文杰[1,2] 邓瑶[2] 管洁[2] 田瑞光[2] 王文[2] 陈红[2] 毕胜利[2] 阮力[2] 

机构地区：[1]中国疾病预防控制中心病毒基因工程国家重点实验室,北京100052 [2]中国疾病预防控制中心病毒病预防控制所,北京100052

出　　处：《中华微生物学和免疫学杂志》2010年第1期41-45,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30671961）;国家863课题资助项目（2007AA02Z455） 志谢：感谢美国NIH VRC的Dr.Nabel惠赠pVRC8301载体质粒.

摘　　要：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.

关 键 词：丙型肝炎病毒 DNA疫苗 体内电转 

分 类 号：R512.63[医药卫生—内科学] R392-33[医药卫生—临床医学]