Center for the Study of Hepatitis C, Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, New York, USA.
Nat Biotechnol. 2010 Feb;28(2):167-71. doi: 10.1038/nbt.1604. Epub 2010 Jan 31.
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, use of genetically modified viruses and/or terminal processing of potentially precious samples. 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.
丙型肝炎病毒(HCV)感染了世界人口的 2-3%,是慢性肝炎的致病因子,也是肝移植的主要指征。在细胞培养中繁殖 HCV(HCVcc)的能力是最近的一个突破,也是寻找特定抗病毒治疗方法的关键工具。在培养物中监测 HCV 感染通常涉及批量群体检测、使用基因修饰病毒和/或对潜在珍贵样本进行末端处理。在这里,我们开发了一种基于细胞的荧光报告系统,可在活细胞或固定样本中灵敏地区分单个 HCV 感染细胞。我们证明了该技术在几个以前难以解决的应用中的用途,包括病毒增殖和宿主反应的活细胞成像,以及可视化原发性肝细胞培养物的感染。将这种报告器与现代基于图像的分析方法相结合,可能为 HCV 研究开辟新的途径。