Department of Gastroenterology and Hepatology Department for Hepatitis Control Department of Advanced Therapeutics in Gastrointestinal Diseases, Tokyo Medical and Dental University Department of Virology II, National Institute of Infectious Disease, Tokyo Department of Internal Medicine, Soka Municipal Hospital, Saitama, Japan.
Hepatol Res. 2011 Mar;41(3):258-69. doi: 10.1111/j.1872-034X.2010.00771.x. Epub 2011 Jan 30.
Studies of the complete hepatitis C virus (HCV) life cycle have become possible with the development of a HCV-JFH1 cell culture system.
In this study, we constructed two fluorescence protein-tagged recombinant JFH1 virus clones, JFH1-EYFP and JFH1-AsRed, as well as two corresponding clones with adaptive mutations, JFH1-EYFP mutant and JFH1-AsRed mutant, that and were as effective as JFH1 in producing infectious virus particles, and investigated their viral infection life cycles.
After infection of the fluorescence-tagged mutant viruses, infected cells increased exponentially. In cells, EYFP or AsRed and NS5A were expressed as a fusion protein and co-localized in core proteins. The rate of the cell-cell spread was dependent on the cell densities with a maximum of 10(2.5) /day. Treatment of cells with interferon or a protease inhibitor suppressed expansion of virus-positive cells.
Taken together, these results indicate that fluorescence-tagged HCV is a useful tool to study virus infection life cycles and to assist in the search for novel antiviral compounds.
随着丙型肝炎病毒（HCV）JFH1 细胞培养系统的发展，对 HCV 完整生命周期的研究成为可能。
本研究构建了两个荧光蛋白标记的重组 JFH1 病毒克隆，JFH1-EYFP 和 JFH1-AsRed，以及两个具有适应性突变的相应克隆，JFH1-EYFP 突变体和 JFH1-AsRed 突变体，它们与 JFH1 一样有效地产生感染性病毒颗粒，并研究了它们的病毒感染生命周期。
感染荧光标记的突变病毒后，受感染的细胞呈指数增长。在细胞中，EYFP 或 AsRed 和 NS5A 作为融合蛋白表达，并与核心蛋白共定位。细胞间传播的速度依赖于细胞密度，最高可达 10(2.5) /天。用干扰素或蛋白酶抑制剂处理细胞可抑制病毒阳性细胞的扩增。
综上所述，这些结果表明荧光标记的 HCV 是研究病毒感染生命周期和辅助寻找新型抗病毒化合物的有用工具。
