流感病毒为载体的HCV疫苗构建及免疫效果研究

HCV is important infectious disease that endanger human health. So far no vaccine is available. To construct HCV C,E1/E2,C+E1/E2 antigen molecular which have B cell neutralizing epitopes and CTL epitopes by using immunological and bioinformatics techniques，chimeric influenza virus vaccines HCV-FLU were generated using reverse genetics in which influenza virus was acted as delivery system which contain six internal backbone from cold-adapted attenuated influenza virus strain A/AA/6/60,NA fragment were targeted for inserting HCV epitopes. Then the safety, immunogenicity and stability of HCV-FLU was evaluated in immunized animals. Sera neutralized infections with HCV pseudoparticles and cell-cultured viruses derived from different heterologous HCV strains or mice challenged with wild-type influenza A virus. Meanwhile the cellular and molecular immune responses mechanism of the recombinant attenuated vaccines were investigeted. This work provides novel powerful tool for HCV vaccine development and theoretic evidence for relizing one kind of vaccine could protect from two infectious disease infection.

丙肝已成为危害人类健康重大传染病，至今没有批准的疫苗使用。本课题采用抗原免疫生物信息学技术，构建具有B 细胞中和抗体和T细胞杀伤效应的HCV 优势抗原表位C、E1/E2、C+E1/E2分子；以冷适应、减毒流感病毒株A/AA/6/60作为递送系统，将NA片段为插入外源基因的靶点，通过反向遗传学技术重配嵌合HCV抗原表位的流感减毒疫苗株HCV-FLU；免疫小鼠实验评价其安全性、免疫原性和稳定性；采集免疫动物血清，与HCV假病毒、感染性克隆细胞培养制备的HCV病毒体外中和试验评价其免疫效果，同时用流感病毒野生株攻毒评价免疫保护效果；进一步探索嵌合HCV-FLU重组减毒疫苗产生免疫应答与保护的细胞、分子机制。本研究不仅为HCV疫苗研究提供新的科学思路，也将为传染病疫苗实现"一苗两用"提供了理论依据。

丙肝已成为危害人类健康重大传染病，至今没有批准的疫苗使用。本课题采用抗原免疫生物信息学技术，构建具有B细胞中和抗体和T细胞杀伤效应的HCV优势抗原表位C、E1/E2、C+E1/E2分子；分别以流感病毒株A/PR/8/34、A/AA/6/60作为递送系统，通过反向遗传学技术重配获得了嵌合HCV抗原表位的流感减毒疫苗株rHCV/FLU、AA/NA/rHCV/FLU；免疫小鼠实验评价其安全性、免疫原性和稳定性；HCV慢性感染病人PBMC的免疫应答反应评价其免疫效果，同时用流感病毒野生株攻毒评价免疫保护效果；进一步探索嵌合HCV-FLU重组减毒疫苗产生免疫应答与保护的细胞、分子机制。本课题的实施和完成，发表及待发表论文6篇，其中SCI 5篇；授权新加坡专利1项，授权国家发明专利2项，申报PCT专利2项；；初步建立了流感病毒载体表达外源抗原的技术平台，为HCV疫苗的研制提供了新思路，同时为呼吸道传染病疫苗实现“一苗两用”或“一苗多用”奠定了理论基础和技术支撑。