Study on molecular mechanisms of hepatitis C virus infection.

丙型肝炎病毒感染的分子机制研究。

• 批准号: 16017252
• 负责人: MATSUURA Yoshiharu
• 金额: $ 9.6万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16017252/
• 关键词: HCV; Infection; Receptor; FGFR; 細胞融合; リセプター

Although several cell surface molecules such as hCD81, LDL-R, SR-B1, and DC-SIGN have been proposed as candidates for hepatitis C virus (HCV) receptor, it is still unclear if any of these molecules can play a role as a functional cellular receptor, due to the lack of robust and reliable in vitro cell culture systems to propagate HCV. As a surrogate system for the study of HCV infection, pseudotype viruses bearing HCV envelope glycoproteins based on vesicular stomatitis viruses (HCVpv) and retroviruses (HCVpp) have been developed. We have previously shown that human fibroblast growth factor receptor (hFGFR) 4 is a binding receptor for HCV based on the specific binding with HCVpv, HCV-like particles and authentic HCV particles in patient sera. Recently we found that HCVpv generated in 293T cells and CHO cells (HCVpv/CHO) exhibit hCD81-dependent and -independent infection, respectively. Infection of HCVpv/CHO to HepG2 cells was inhibited by hFGF2, a soluble protein representing the ectodomain of hFGFR5 fused with the Fe region of IgG (hFGFR5/Fc), or anti-hFGFR5 antibody. Overexpression of hFGFR5 in 293T and Huh7 cells enhanced the susceptibility to HCVpv/CHO infection. In contrast, siRNA-mediated knockdown of hFGFR5 in HepG2 cells resulted in the reduction of infectivity of HCVpv/CHO. Furthermore, binding of the authentic HCV particles in patient sera to HepG2 cells was inhibited by the hFGFR5/Fc. Finally, expression of hFGFR5 (but not hFGFR4) in CHO cells rendered them permissive for HCVpv/CHO infection. Together, these results suggest the possible involvement of hFGFR5 in the internalization process of HCV in a hCD81-independent fashion.

尽管 hCD81、LDL-R、SR-B1 和 DC-SIGN 等几种细胞表面分子已被提议作为丙型肝炎病毒 (HCV) 受体的候选分子，但仍不清楚这些分子中的任何一个是否可以发挥丙型肝炎病毒 (HCV) 受体的作用。功能性细胞受体，由于缺乏强大且可靠的体外细胞培养系统来传播 HCV。作为研究 HCV 感染的替代系统，基于水泡性口炎病毒 (HCVpv) 和逆转录病毒 (HCVpp) 的带有 HCV 包膜糖蛋白的假型病毒已被开发出来。我们之前已经证明，人成纤维细胞生长因子受体（hFGFR）4是HCV的结合受体，基于与患者血清中的HCVpv、HCV样颗粒和真实HCV颗粒的特异性结合。最近我们发现293T细胞和CHO细胞(HCVpv/CHO)中产生的HCVpv分别表现出hCD81依赖性和非hCD81依赖性感染。 HCVpv/CHO 对 HepG2 细胞的感染被 hFGF2 或抗 hFGFR5 抗体抑制，hFGF2 是一种代表与 IgG Fe 区融合的 hFGFR5 胞外域的可溶性蛋白（hFGFR5/Fc）。 293T 和 Huh7 细胞中 hFGFR5 的过度表达增强了对 HCVpv/CHO 感染的易感性。相反，HepG2细胞中siRNA介导的hFGFR5敲低导致HCVpv/CHO感染性降低。此外，患者血清中的真实HCV颗粒与HepG2细胞的结合被hFGFR5/Fc抑制。最后，CHO 细胞中 hFGFR5（但不是 hFGFR4）的表达使其允许 HCVpv/CHO 感染。总之，这些结果表明 hFGFR5 可能以不依赖 hCD81 的方式参与 HCV 的内化过程。

项目成果

Intramembrane proteolysis and ER retention of HCV core protein.

HCV 核心蛋白的膜内蛋白水解和 ER 保留。

• 发表时间: 2004
• 期刊: Journal of Virology 78
• 作者: Shimokata K;Yamada Y;Konda T;Izawa H;Nagata K;Murohara T;Ohno M;Yokota M;Okamoto K.
• 通讯作者: Okamoto K.

Identification of basal promoter and enhancer elements in an untranslated reeion of the Ti' virus eenome.

Ti病毒基因组非翻译区中基础启动子和增强子元件的鉴定。

• 发表时间: 2004
• 期刊: J. Virol. 78
• 作者: Suzuki T.;Suzuki H.;Hiplcata M.;Matsuda M.;Li T-C.;Matsuura Y.;Mishiro S.;Miyamura T.
• 通讯作者: Miyamura T.

Human VAP-B Is Involve in Hapatitis C Virus Replication through Interaction with NS5A and NS5B.

人 VAP-B 通过与 NS5A 和 NS5B 相互作用参与丙型肝炎病毒复制。

• 发表时间: 2005
• 期刊: Journal of Virology 79
• 作者: Tsuda Y.;Mori Y.;Abe T.;Yamashita T.;Okamoto T.;Ichimura T.;Moriishi K;Matsuura Y.;Hamanoto I.
• 通讯作者: Hamanoto I.

Evidence for an alternative topology of the E1 envelope glycoprotein of hepatitis C virus.

丙型肝炎病毒 E1 包膜糖蛋白替代拓扑结构的证据。

• 发表时间: 2004
• 期刊: Virus Research 105
• 作者: Li T.C.;Takeda N.;Miyamura T.;Matsuura Y.;Wang J.C.;Engvall H.;Hammar L;Xing L;Cheng R.H.;Hamamoto I.;Okamoto K.;Kaimori A.;Migliaccio C.T.
• 通讯作者: Migliaccio C.T.

Essential elements of the capsid protein for self-assembly into empty virus-like particles of hapatitis E virus.

衣壳蛋白自组装成戊型肝炎病毒空病毒样颗粒的基本要素。

• 发表时间: 2005
• 期刊: Journal of Virology 79
• 作者: Tsuda Y.;Mori Y.;Abe T.;Yamashita T.;Okamoto T.;Ichimura T.;Moriishi K;Matsuura Y.;Hamanoto I.;Abe T.;Kitagawa Y.;Mori Y.;Li T.C.
• 通讯作者: Li T.C.