Ezelle Heather J, Markovic Dubravka, Barber Glen N
Department of Microbiology and Immunology and Sylvester Comprehensive Cancer Center, University of Miami School of Medicine, Miami, Florida 33136, USA.
J Virol. 2002 Dec;76(23):12325-34. doi: 10.1128/jvi.76.23.12325-12334.2002.
Hepatitis C virus (HCV), a major etiologic agent of hepatocellular carcinoma, presently infects approximately 400 million people worldwide, making the development of protective measures against HCV infection a key objective. Here we have generated a recombinant vesicular stomatitis virus (VSV), which expresses the HCV structural proteins, by inserting the contiguous Core, E1, and E2 coding region of HCV into the VSV genome. Recombinant VSV expressing HCV Core, E1, and E2 (VSV-HCV-C/E1/E2) grew to high titers in vitro and efficiently expressed the incorporated HCV gene product, which became fully processed into the individual HCV structural proteins. Biochemical and biophysical analysis indicated that the HCV Core, E1, and E2 proteins assembled to form HCV-like particles (HCV-LPs) possessing properties similar to the ultrastructural properties of HCV virions. Mice immunized with VSV-HCV-C/E1/E2 generated cell-mediated immune responses to all of the HCV structural proteins, and humoral responses, particularly to E2, were also readily evident. Our data collectively indicate that engineered VSVs expressing HCV Core, E1, and E2 and/or HCV-LPs represent useful tools in vaccine and immunotherapeutic strategies designed to address HCV infection.
丙型肝炎病毒（HCV）是肝细胞癌的主要病因，目前全球约有4亿人感染该病毒，因此开发针对HCV感染的防护措施成为一项关键目标。在此，我们通过将HCV连续的核心蛋白、E1蛋白和E2蛋白编码区插入水泡性口炎病毒（VSV）基因组，构建了一种表达HCV结构蛋白的重组水泡性口炎病毒（VSV）。表达HCV核心蛋白、E1蛋白和E2蛋白的重组VSV（VSV-HCV-C/E1/E2）在体外能达到高滴度，并能有效表达整合的HCV基因产物，该产物能完全加工成单个的HCV结构蛋白。生化和生物物理分析表明，HCV核心蛋白、E1蛋白和E2蛋白组装形成了具有类似于HCV病毒粒子超微结构特性的HCV样颗粒（HCV-LPs）。用VSV-HCV-C/E1/E2免疫的小鼠对所有HCV结构蛋白产生了细胞介导的免疫反应，并且体液免疫反应，特别是针对E2蛋白的反应也很明显。我们的数据共同表明，表达HCV核心蛋白、E1蛋白和E2蛋白的工程化VSV和/或HCV-LPs是疫苗和免疫治疗策略中用于应对HCV感染的有用工具。
