Studies on virus kinetics using infectious fluorescence-tagged hepatitis C virus cell culture
Studies on virus kinetics using infectious fluorescence-tagged hepatitis C virus cell culture
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DOI：
10.1111/j.1872-034x.2010.00771.x
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发表时间：
2011-03-01
影响因子：
4.2
通讯作者：
Watanabe, Mamoru
中科院分区：
文献类型：
--
作者：
Yamamoto, Machi;Sakamoto, Naoya;Watanabe, Mamoru
目的：随着HCV- jfh1细胞培养系统的发展，研究完整的丙型肝炎病毒（HCV）生命周期成为可能。方法：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，以及对应的两个具有适应性突变的克隆JFH1- eyfp突变体和JFH1- asred突变体，它们在产生感染性病毒颗粒方面与JFH1一样有效，并研究了它们的病毒感染生命周期。结果：荧光标记突变病毒感染后，感染细胞呈指数增长。在细胞中，EYFP或AsRed和NS5A作为融合蛋白表达，并在核心蛋白中共定位。细胞-细胞扩散速率与细胞密度有关，最大可达102.5/d。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。结论：综上所述，这些结果表明荧光标记HCV是研究病毒感染生命周期和协助寻找新的抗病毒化合物的有用工具。
Aim:Studies of the complete hepatitis C virus (HCV) life cycle have become possible with the development of a HCV-JFH1 cell culture system.Methods: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.Results: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 102.5/day. Treatment of cells with interferon or a protease inhibitor suppressed expansion of virus-positive cells.Conclusion: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.