Production of Hepatitis C Virus Lacking the Envelope-Encoding Genes for Single-Cycle Infection by Providing Homologous Envelope Proteins or Vesicular Stomatitis Virus Glycoproteins in trans
Production of Hepatitis C Virus Lacking the Envelope-Encoding Genes for Single-Cycle Infection by Providing Homologous Envelope Proteins or Vesicular Stomatitis Virus Glycoproteins in trans
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DOI：
10.1128/jvi.02313-10
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发表时间：
2010-12
影响因子：
5.4
通讯作者：
Rui Li;Yan Qin;Ying He;Wanyin Tao;N. Zhang;Cheguo Tsai;P. Zhou;Jin Zhong
中科院分区：
文献类型：
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作者：
Rui Li;Yan Qin;Ying He;Wanyin Tao;N. Zhang;Cheguo Tsai;P. Zhou;Jin Zhong
丙型肝炎病毒（HCV）感染是一个世界性的主要健康问题。包膜糖蛋白是病毒颗粒的主要成分。在这里，我们开发了一个反式互补系统，允许生产感染性HCV颗粒，其基因组中编码包膜蛋白的区域被删除（HCVΔE）。在反式中表达同源包膜蛋白可以有效地弥补包膜蛋白的缺失。HCVΔE的产生可以通过先前描述的NS3和NS5A的适应性突变显著增强。此外，HCVΔE可以在表达HCV包膜蛋白的包装细胞中稳定地繁殖传代，在野生型细胞中仅产生单轮感染。有趣的是，我们发现水疱性口炎病毒（VSV）糖蛋白可以有效地挽救缺乏内源性包膜蛋白的HCV的产生，而内源性包膜蛋白不再需要载脂蛋白E来产生病毒。VSV糖蛋白介导的病毒进入可以绕过自然的HCV进入过程，并将HCV复制子RNA递送到HCV受体缺陷细胞中。我们的研究为单循环感染性HCV颗粒的生产提供了一种新的工具，这将有助于研究HCV生命周期的各个步骤，也可能为HCV疫苗的开发提供新的策略。
ABSTRACT Hepatitis C virus (HCV) infection is a major worldwide health problem. The envelope glycoproteins are the major components of viral particles. Here we developed a trans-complementation system that allows the production of infectious HCV particles in whose genome the regions encoding envelope proteins are deleted (HCVΔE). The lack of envelope proteins could be efficiently complemented by the expression of homologous envelope proteins in trans. HCVΔE production could be enhanced significantly by previously described adaptive mutations in NS3 and NS5A. Moreover, HCVΔE could be propagated and passaged in packaging cells stably expressing HCV envelope proteins, resulting in only single-round infection in wild-type cells. Interestingly, we found that vesicular stomatitis virus (VSV) glycoproteins could efficiently rescue the production of HCV lacking endogenous envelope proteins, which no longer required apolipoprotein E for virus production. VSV glycoprotein-mediated viral entry could allow for the bypass of the natural HCV entry process and the delivery of HCV replicon RNA into HCV receptor-deficient cells. Our development provides a new tool for the production of single-cycle infectious HCV particles, which should be useful for studying individual steps of the HCV life cycle and may also provide a new strategy for HCV vaccine development.