Castet Valérie, Moradpour Darius
Department of Medicine II, University of Freiburg, Freiburg, Germany.
Hepatology. 2003 Sep;38(3):771-4. doi: 10.1002/hep.510380328.
The study of hepatitis C virus (HCV), a major cause of chronic liver disease, has been hampered by the lack of a cell culture system supporting its replication. Here, we have successfully generated infectious pseudo-particles that were assembled by displaying unmodified and functional HCV glycoproteins onto retroviral and lentiviral core particles. The presence of a green fluorescent protein marker gene packaged within these HCV pseudo-particles allowed reliable and fast determination of infectivity mediated by the HCV glycoproteins. Primary hepatocytes as well as hepato-carcinoma cells were found to be the major targets of infection in vitro. High infectivity of the pseudo-particles required both E1 and E2 HCV glycoproteins, and was neutralized by sera from HCV-infected patients and by some anti-E2 monoclonal antibodies. In addition, these pseudo-particles allowed investigation of the role of putative HCV receptors. Although our results tend to confirm their involvement, they provide evidence that neither LDLr nor CD81 is sufficient to mediate HCV cell entry. Altogether, these studies indicate that these pseudo-particles may mimic the early infection steps of parental HCV and will be suitable for the development of much needed new antiviral therapies.
丙型肝炎病毒（HCV）是慢性肝病的主要病因，对其研究因缺乏支持病毒复制的细胞培养系统而受阻。在此，我们成功构建了感染性假病毒颗粒，该颗粒通过将未修饰的功能性HCV糖蛋白展示在逆转录病毒和慢病毒核心颗粒上组装而成。这些HCV假病毒颗粒中包装的绿色荧光蛋白标记基因使得能够可靠且快速地测定由HCV糖蛋白介导的感染性。原代肝细胞以及肝癌细胞被发现是体外感染的主要靶细胞。假病毒颗粒的高感染性需要E1和E2两种HCV糖蛋白，并且能被HCV感染患者的血清以及一些抗E2单克隆抗体中和。此外，这些假病毒颗粒有助于研究推定的HCV受体的作用。尽管我们的结果倾向于证实它们的参与，但结果表明LDLr和CD81均不足以介导HCV进入细胞。总之，这些研究表明这些假病毒颗粒可能模拟亲本HCV的早期感染步骤，将适用于开发急需的新型抗病毒疗法。
