HEPATITIS C--MODELS FOR REPLICATION

丙型肝炎病毒（HCV）是慢性肝炎的主要原因， 肝衰竭和癌症的并发症。 前景 开发一种有效的疫苗仍然是不可能的， 新的抗病毒药物由于缺乏HCV模型系统而受到限制。 的 对选定的慢性乙型肝炎患者人群的进一步表征 HCV，以及研究HCV复制所需的模型系统的开发 并确定基于分子的治疗方法和模型的开发 研究HCV复制和识别基于分子的 治疗学代表了主要的未解决的问题。 HCV RNA依赖性RNA 聚合酶（RDRP）是HCV抗病毒药物的可能靶点。 我们已经表示 重组HCV RDRP，并将其作为活性酶分离。 另外我们 已经在一些人中检测到了针对重组HCV RDRP的循环抗体， 患者-这可能提供病毒复制的另一个指标。 此外，委员会认为， 我们已经在开发一个系统方面取得了进展， 培养细胞 待检验的假设是：体外模型和 可以开发细胞系统来研究HCV复制的关键方面， 并确定直接靶向HCV的分子疗法 RNA依赖性RNA聚合酶。 具体目标： 1)纯化和鉴定重组HCV RNA依赖性RNA聚合酶 （RDRP），我们在E.大肠杆菌，并已分离出一个活跃的 形式; 2）使用我们的HCV RDRP检测来鉴定 直接抑制这种酶。 3. 用纯化的HCV RDRP进行 将用于进一步表征患者的免疫测定， 慢性丙型肝炎，以及4）为未来晶体做可行性研究 成长工作。 使用表达活性HCV RNA依赖性RA聚合酶的基因， 开发模型细胞系统，将允许进一步研究这一点， 酶和识别小分子，进入完整的细胞，并抑制 HCV RDRP。 继续研究一种有前途的细胞培养系统， 小鼠研究。 我们的长期目标是利用这个系统来研究 病毒基因表达和复制的详细信息，并确定移动的HCV 抗病毒剂。

Hepatitis C virus (HCV) is a major cause of chronic hepatitis, with complications of liver failure and cancer, throughout the world. Prospects for developing an effective vaccine remain unlikely, and the development of new antivirals has been limited by the lack of model HCV systems. The further characterization of selected populations of patients with chronic HCV, and the development of model systems needed to study HCV replication and to identify molecular-based therapeutics and the development of model systems needed to study HCV replication and to identify molecular-based therapeutics represents major unsolved problems. The HCV RNA-dependent RNA polymerase (RDRP) is a likely target for HCV antivirals. We have expressed recombinant HCV RDRP an isolated it as an active enzyme. In addition, we have detected circulating antibodies to recombinant HCV RDRP in some patients - this may provide another index of viral replication. Moreover, we have made progress in developing a system that propagates HCV in cultured cells. The hypothesis to be tested is: that model in vitro and cellular systems can be developed to study key aspects of HCV replication, and to identify molecular-based therapeutics that directly target the HCV RNA-dependent RNA polymerase. Specific Objectives: 1) to purify and characterize recombinant HCV RNA-dependent RNA polymerase (RDRP) that we are expressing in E. coli and have isolated in an active form; 2) to use our HCV RDRP assay to identify small molecules that directly inhibit this enzyme. 3. To use purified HCV RDRP to perform immunoassays that will be used to further characterize patients with chronic hepatitis C, and 4) to do feasibility studies for future crystal growth work. The use the gene that expresses active HCV RNA=dependent RA polymerase to develop model cellular systems that will permit further studies of this enzyme and identify small molecules that enter intact cells and inhibit the HCV RDRP. To continue studies on a promising cell culture system that propagates HCV mouse studies. Our long-range goal will be to use this system to study details of viral gene expression and replication, and to identify moved HCV antiviral agents.