原钙黏蛋白β5介导丙型肝炎病毒入胞的作用及机制研究

项目名称： 原钙黏蛋白β5介导丙型肝炎病毒入胞的作用及机制研究

项目编号： No.81201291

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学四处

项目作者： 朱海红

作者单位： 浙江大学

项目金额： 23万元

中文摘要： 全球HCV感染者约为1.7亿，目前尚无预防用疫苗,因此HCV感染是日益严重的公共卫生问题。HCV入胞相关宿主因子是潜在药物靶点。近年来随着HCV体外细胞培养体系的突破性进展，多个参与HCV入胞的宿主因子的发现使HCV入胞过程逐渐清晰，但尚未完全明了。本课题前期工作发现细胞黏附分子- - 原钙黏蛋白β5（PCDHB5）可介导HCV的入胞过程。本申请拟构建不同截短长度PCDHB5表达质粒，研究PCDHB5与HCV E2的结合功能域；分别共转染PCDHB5和已知HCV入胞相关分子CD81/SR-B1/CLDN1/OCLN的表达质粒，用免疫共沉淀研究PCDHB5与这些相关分子的相互作用；制备抗PCDHB5单克隆抗体，筛选具有抑制不同基因型HCV入胞功能的高亲和力的中和抗体。深入研究PCDHB5介导HCV入胞作用及机制，以进一步解析HCV生活周期，为全面阐明HCV致病机制和研制新的防治策略提供理论依据。

中文关键词： 丙型肝炎病毒；原钙黏蛋白；病毒入胞；细胞黏附分子；受体

英文摘要： Hepatitis C virus (HCV) infection represents a major public health threat with a world-wide 170 million peoples infected and no vaccine available for prevention. Viral entry represents a promising target for antiviral intervention. More recently, a cell culture system that supports the production and propagation of infectious HCV in cell culture (HCVcc) has been developed. Several host cell receptors have been proposed to be involved in the viral entry. Our preliminary studies discovered protocadherin beta 5 (PCDHB5), a cellular adhesion molecule, as a cellular co-factor for HCV entry. In this proposal, we plan to further discover the role of PCDHB5 in HCV entry, and generate a neutralizing monoclonal antibody for future in vivo studies to address the feasibility of an antibody therapeutic. First, the functional domains of PCDHB5 involved in conferring HCV infectivity and E2 binding will be mapped. The specific interaction of PCDHB5 with HCV E2 may be part of the mechanism for its ability to confer infectivity. It would be important to identify specific regions of the respective proteins involved or the nature of the interactions, as well as to determine if such interaction can induce down-stream signaling that is important for virus infection. Different truncated PCDHB5 expression plasmids will be constructed

英文关键词： Hepatitis c virus；protocadherin beta 5；virus entry；cellular adhesion molecule；receptor