Ait-Goughoulte Malika, Kanda Tatsuo, Meyer Keith, Ryerse Jan S, Ray Ratna B, Ray Ranjit
Division of Infectious Diseases and Immunology, Center for Vaccine Development, Edward A. Doisy Research Center, 1100 S. Grand Blvd., 8th Floor, St. Louis, MO 63104, USA.
J Virol. 2008 Mar;82(5):2241-9. doi: 10.1128/JVI.02093-07. Epub 2007 Dec 12.
We have previously reported that immortalized human hepatocytes (IHH) support the generation of infectious hepatitis C virus (HCV) genotype 1a (clone H77). In the present study, we have investigated the growth of HCV genotype 1a (clone H77) through serial passages and accompanying changes in IHH in response to infection. Eleven serial passages of HCV genotype 1a (clone H77) in IHH were completed. Virus replication was ascertained from the presence of HCV-specific sequences, the detection of core antigen, the virus genome copy number, and the virus titer in IHH culture fluid. Electron microscopy suggested that HCV infection induces autophagic vacuole formation in IHH. Fluorescence microscopy displayed localization of autophagic markers, microtubule-associated protein-1 light chain-3 and Apg5, on the vacuoles of HCV-infected hepatocytes. Taken together, our results suggested that HCV genotype 1a (clone H77) can be serially passaged in IHH and that HCV infection induces an autophagic response in hepatocytes.
我们之前报道过，永生化人肝细胞（IHH）支持传染性丙型肝炎病毒（HCV）1a基因型（克隆H77）的产生。在本研究中，我们通过连续传代研究了HCV 1a基因型（克隆H77）的生长情况以及IHH中伴随感染发生的变化。完成了HCV 1a基因型（克隆H77）在IHH中的11次连续传代。通过HCV特异性序列的存在、核心抗原的检测、病毒基因组拷贝数以及IHH培养液中的病毒滴度来确定病毒复制情况。电子显微镜检查表明，HCV感染可诱导IHH中自噬泡的形成。荧光显微镜显示，自噬标记物微管相关蛋白1轻链3和Apg5定位于HCV感染的肝细胞的液泡上。综合来看，我们的结果表明，HCV 1a基因型（克隆H77）可在IHH中连续传代，并且HCV感染可诱导肝细胞发生自噬反应。
