Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
Laboratory of Veterinary Microbiology, School of Veterinary Medicine, Kitasato University, Aomori, Japan.
Sci Rep. 2017 Mar 22;7:45228. doi: 10.1038/srep45228.
It is well documented that a variety of viral quasispecies are found in the patients with chronic infection of hepatitis C virus (HCV). However, the significance of quasispecies in the specific infectivity to individual cell types remains unknown. In the present study, we analyzed the role of quasispecies of the genotype 2a clone, JFH1 (HCVcc), in specific infectivity to the hepatic cell lines, Huh7.5.1 and Hep3B. HCV RNA was electroporated into Huh7.5.1 cells and Hep3B/miR-122 cells expressing miR-122 at a high level. Then, we adapted the viruses to Huh7 and Hep3B/miR-122 cells by serial passages and termed the resulting viruses HCVcc/Huh7 and HCVcc/Hep3B, respectively. Interestingly, a higher viral load was obtained in the homologous combination of HCVcc/Huh7 in Huh7.5.1 cells or HCVcc/Hep3B in Hep3B/miR-122 cells compared with the heterologous combination. By using a reverse genetics system and deep sequence analysis, we identified several adaptive mutations involved in the high affinity for each cell line, suggesting that quasispecies of HCV participate in cell-specific infectivity.
有大量文献记载表明，慢性丙型肝炎病毒（HCV）感染者体内存在多种病毒准种。然而，病毒准种在对特定细胞类型的特异性感染中的作用仍不清楚。本研究分析了基因型 2a 克隆 JFH1（HCVcc）的准种在对 Hep3B/miR-122 细胞和 Huh7.5.1 细胞这两种肝系细胞株的特异性感染中的作用。我们用电穿孔的方法将 HCV RNA 转染到高表达 miR-122 的 Hep3B/miR-122 细胞和 Huh7.5.1 细胞中，然后通过连续传代使病毒适应 Huh7 细胞和 Hep3B/miR-122 细胞，并分别将得到的病毒命名为 HCVcc/Huh7 和 HCVcc/Hep3B。有趣的是，与异源组合相比，HCVcc/Huh7 在 Huh7.5.1 细胞或 HCVcc/Hep3B 在 Hep3B/miR-122 细胞中的同源组合中获得了更高的病毒载量。通过使用反向遗传学系统和深度测序分析，我们鉴定出了几个与每种细胞系高亲和力相关的适应性突变，提示 HCV 的病毒准种参与了细胞特异性感染。
