Analysis of virus adaptations in the novel HCV cell culture and infection system

新型 HCV 细胞培养和感染系统中病毒适应性分析

• 批准号: 18590462
• 负责人: MURAYAMA Asako
• 金额: $ 2.57万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590462/
• 关键词: HCV; Virus; Virus replication; adaptation; Infection; ウイルスゲノム複製; 変異; 慢性肝炎; 肝臓癌

We have established HCV culture system using JFH-1 strain isolated from a fulminant hepatitis patient. This system enabled us to analyze HCV life cycles in detail. In this research project, we have analyzed how HCV adapted in cultured cells and organisms. We have also tried to determine which virus components involved in HCV adaptation.HCV adaptation in cultured cells: we transfected in vitro-synthesized HCV RNA into Huh7 cells and collected infectious HCV particles. HCV infection sometimes causes plaque formation. We collected plaque forming HCVs, and infected to naive Huh7 cells. Isolated virus subclones from individual plaque showed greater replication efficiency and cytopathogenicity than parental virus. We analyzed the sequence of this HCV and found several nucleotide mutations with amino acid substitution in NS5B coding region. These mutations may be involved in plaque forming ability and virus genome replication.

我们用从一例重型肝炎患者中分离的JFH-1株建立了丙型肝炎病毒的培养体系。这个系统使我们能够详细分析丙型肝炎病毒的生命周期。在这个研究项目中，我们分析了丙型肝炎病毒如何在培养的细胞和生物体中适应。我们还试图确定哪些病毒成分参与了丙型肝炎病毒的适应。培养细胞中的丙型肝炎适应：我们将体外合成的丙型肝炎病毒RNA导入Huh7细胞，并收集具有感染性的丙型肝炎病毒颗粒。丙型肝炎病毒感染有时会导致斑块形成。收集形成丙型肝炎病毒的空斑，感染未成熟的HuH7细胞。从单个斑块中分离的病毒亚克隆显示出比亲本病毒更高的复制效率和细胞致病性。我们分析了该丙型肝炎病毒的序列，发现在NS5B编码区有几个氨基酸替换的核苷酸突变。这些突变可能与空斑形成能力和病毒基因组复制有关。

项目成果

C型肝炎ウイルス脂質成分の感染における役割

丙型肝炎病毒脂质成分在感染中的作用

• 发表时间: 2007
• 作者: 相崎 英樹、鈴木 哲朗;他
• 通讯作者: 他

HCV research and anti-HCV drug discovery: toward the next generation.

• DOI: 10.1016/j.addr.2007.08.021
• 发表时间: 2007-10
• 期刊: Advanced drug delivery reviews
• 影响因子: 16.1
• 作者: T. Wakita

レジデントノート

居民笔记

• 发表时间: 2007
• 作者: K Morikawa;T Wakita. Infectious hepatitis C virus particle binding to the Huh7 cell surface is mediated by glycosaminoglycans;its internalization by CD81.;森川賢一
• 通讯作者: 森川賢一

Development of plaque assays for hepatitis C virus-JFH1 strain and isolation of mutants with enhanced cytopathogenicity and replication capacity.

• DOI: 10.1016/j.virol.2007.09.019
• 发表时间: 2008-02
• 期刊: Virology
• 影响因子: 3.7
• 作者: Yuko Sekine‐Osajima;N. Sakamoto;Kako Mishima;M. Nakagawa;Yasuhiro Itsui;M. Tasaka;Yuki Nishimura‐Sakurai;Cheng-Hsin Chen;T. Kanai;K. Tsuchiya;T. Wakita;N. Enomoto;Mamoru Watanabe

Subcellular localization of hepatitis C virus structural proteins in a cell culture system that efficiently replicates the virus

• DOI: 10.1128/jvi.80.6.2832-2841.2006
• 发表时间: 2006-03-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Rouillé, Y;Helle, F;Dubuisson, J
• 通讯作者: Dubuisson, J