Cell culture and infection system for hepatitis C virus
Cell culture and infection system for hepatitis C virus
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DOI：
10.1038/nprot.2006.395
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Wakita, Takaji
中科院分区：
文献类型：
--
作者：
Kato, Takanobu;Date, Tomoko;Wakita, Takaji
丙型肝炎病毒（HCV）感染会导致慢性肝脏疾病，是一个全球性的健康问题。尽管对病毒复制和发病机制的了解需求日益增加，但由于缺乏有效的病毒培养系统，详细分析受到了阻碍。我们从一名暴发性肝炎患者体内分离出了丙型肝炎病毒2a基因型JFH - 1株。该毒株可在Huh7细胞中高效复制。通过将体外转录的全长JFH - 1 RNA转染到Huh7细胞中，可在细胞培养中实现重组病毒颗粒的高效复制和分泌。在细胞培养中产生的JFH - 1病毒对未感染过的Huh7细胞和黑猩猩都具有感染性。在易感细胞系中，病毒产生的效率和所产生病毒的感染性都有显著提高。本实验方案描述了如何使用该系统，它为研究病毒生命周期、构建抗病毒策略以及开发有效疫苗提供了有力工具。按照本方案，12天即可获得病毒颗粒。
Hepatitis C virus (HCV) infection causes chronic liver disease and is a worldwide health problem. Despite ever-increasing demand for knowledge on viral replication and pathogenesis, detailed analysis has been hampered by a lack of efficient viral culture systems. We isolated HCV genotype 2a strain JFH-1 from a patient with fulminant hepatitis. This strain replicates efficiently in Huh7 cells. Efficient replication and secretion of recombinant viral particles can be obtained in cell culture by transfection of in vitro - transcribed full-length JFH-1 RNA into Huh7 cells. JFH-1 virus generated in cell culture is infectious for both naive Huh7 cells and chimpanzees. The efficiency of viral production and infectivity of generated virus is substantially improved with permissive cell lines. This protocol describes how to use this system, which provides a powerful tool for studying viral life cycle and for the construction of antiviral strategies and the development of effective vaccines. Viral particles can be obtained in 12 days with this protocol.