Modeling GBV-B Replication in Immortalized Cell Lines

描述(由申请人提供)：目前尚无可供丙型肝炎病毒(丙型肝炎病毒)复制的强大细胞培养系统。因此，丙型肝炎病毒复制的代理模型经常用于抗病毒化合物的评估。GB病毒B(GBV-B)是与丙型肝炎病毒亲缘关系最近的病毒，是丙型肝炎病毒药物开发的有吸引力的替代模型。不幸的是，到目前为止，GBV-B只能在取自小型非人灵长类动物的原代肝细胞中生长。这使得目前的GBV-B抗病毒检测难以进行、昂贵，并且不适用于高通量(HTP)筛查。这项应用旨在利用永生化细胞系开发一种稳健的GBV-B复制细胞培养模型。该项目的具体目标是使GBV-B易感动物的原代肝细胞永生化，鉴定这些细胞系和其他已建立的细胞系是否能够支持GBV-B体外复制，并标准化和验证用于抗病毒药物评估的模型系统。该项目将使GBV-B抗病毒试验广泛可用，负担得起，并适用于HTP应用。这将促进新的抗丙型肝炎病毒药物的发现和开发，这些药物可能会改善丙型肝炎病毒感染者的生活质量。

DESCRIPTION (provided by applicant): There is no robust cell culture system in which the hepatitis C virus (HCV) can replicate. As such, surrogate models of HCV replication are frequently used for the evaluation of antiviral compounds. The GB virus B (GBV-B) is the closest related virus phylogenetically with HCV and is an attractive surrogate model for HCV drug development efforts. Unfortunately, GBV-B can so far only be grown in primary hepatocytes taken from small non-human primates. This makes the current GBV-B antiviral assay difficult to perform, expensive, and not amenable to high-throughput (HTP) screening. This application aims to develop a robust cell culture model of GBV-B replication utilizing immortalized cell lines. The specific aims of this project are to immortalize the primary hepatocytes of GBV-B-susceptible animals, identify if these and other established cell lines can support GBV-B replication in vitro, and to standardize and validate the model system produced for use in the evaluation of antiviral agents. This project will make the GBV-B antiviral assay widely available, affordable, and appropriate for HTP applications. This will facilitate the discovery and development of new anti-HCV drugs that may improve the quality of life for HCV-infected patients.