Real-time imaging of hepatitis C virus infection using a fluorescent cell-based reporter system.

Real-time imaging of hepatitis C virus infection using a fluorescent cell-based reporter system.
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DOI:
10.1038/nbt.1604
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发表时间:
2010-02
影响因子:
46.9
通讯作者:
--
中科院分区:
工程技术1区
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丙型肝炎病毒(HCV)感染世界人口的2-3%,是慢性肝炎的病原体,也是肝移植的主要适应症。在细胞培养中繁殖HCV的能力(HCV-HCV)是一个相对较新的突破,也是寻求特异性抗病毒治疗的关键工具。监测培养物中的HCV感染通常涉及大量群体测定和/或潜在珍贵样品的最终处理。活细胞成像避免了这一点,但需要基因修饰的报告病毒,这往往表现出深刻的复制缺陷。在这里,我们开发了一种基于细胞的荧光报告系统,允许活的或固定的样品中的单个HCV感染的细胞的敏感地区分。我们证明了使用这种技术的几个以前棘手的应用,包括活细胞成像的病毒繁殖和宿主反应,以及可视化感染的原代肝细胞培养。该报告与现代图像分析方法的结合可能为HCV研究打开新的大门。
Hepatitis C virus (HCV), which infects 2-3% of the world population, is a causative agent of chronic hepatitis and the leading indication for liver transplantation. The ability to propagate HCV in cell culture (HCVcc) is a relatively recent breakthrough, and a key tool in the quest for specific antiviral therapeutics. Monitoring HCV infection in culture generally involves bulk population assays and/or terminal processing of potentially precious samples. Live-cell imaging avoids this, but necessitates genetically modified reporter viruses, which often exhibit profound replication defects. Here we develop a cell-based fluorescent reporter system that allows sensitive distinction of individual HCV-infected cells in live or fixed samples. We demonstrate use of this technology for several previously intractable applications, including live-cell imaging of viral propagation and host response, as well as visualizing infection of primary hepatocyte cultures. Integration of this reporter with modern image-based analysis methods could open new doors for HCV research.
DOI: 10.1073/pnas.0504877103
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影响因子: 11.1
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