Development of a flow cytometry live cell assay for the screening of inhibitors of hepatitis C virus (HCV) replication.

The Open Virology Journal Pub Date : 2012-01-01 Epub Date: 2012-11-26 DOI:10.2174/1874357901206010097
Jose A Garcia-Rivera, Kai Lin, Sam Hopkins, Matthew A Gregory, Barrie Wilkinson, Philippe A Gallay
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引用次数: 5

Abstract

In this study, we established a flow cytometry live cell-based assay that permits the screening of hepatitis C virus (HCV) inhibitors. Specifically, we created a stable cell line, which harbors a subgenomic replicon encoding an NS5A-YFP fusion protein. This system allows direct measurement of YFP fluorescence in live hepatoma cells in which the HCV replicon replicates. We demonstrated that this stable fluorescent system permits the rapid and sensitive quantification of HCV replication inhibition by direct-acting antiviral agents (DAA) including protease and NS5A inhibitors and host-targeting antiviral agents (HTA) including cyclophilin inhibitors. This flow cytometry-based live cell assay is well suited for multiple applications such as the evaluation of HCV replication as well as antiviral drug screening.

Abstract Image

Abstract Image

用于筛选丙型肝炎病毒(HCV)复制抑制剂的流式细胞术活细胞试验的发展。
在这项研究中,我们建立了一种基于流式细胞术的活细胞检测方法,可以筛选丙型肝炎病毒(HCV)抑制剂。具体来说,我们创建了一个稳定的细胞系,其中包含一个编码NS5A-YFP融合蛋白的亚基因组复制子。该系统允许在HCV复制子复制的活肝癌细胞中直接测量YFP荧光。我们证明,这种稳定的荧光系统允许快速和敏感地定量直接作用抗病毒药物(DAA),包括蛋白酶和NS5A抑制剂和宿主靶向抗病毒药物(HTA),包括亲环蛋白抑制剂抑制HCV复制。这种基于流式细胞术的活细胞检测非常适合多种应用,如HCV复制的评估以及抗病毒药物筛选。
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