Development of plaque assay to isolate cytopathic HCV clones

开发噬菌斑测定法来分离细胞病变的 HCV 克隆

• 批准号: 21590832
• 负责人: OOOKA Shinya
• 金额: $ 2.91万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21590832/
• 关键词: HCVレプリコン; HCV-JFH1培養系; プラークアッセイ; 細胞障害効果; ヒト肝臓キメラマウス; 小胞体ストレス; RNAポリメラーゼ

HCV-JFH1 yields subclones that develop cytopathic plaques(Sekine-Osajima Y, et al., Virology 2008 ; 371 : 71). Here, we investigated viral amino acid substitutions in cytopathic mutant HCV-JFH1 clones and their characteristics in vitro and in vivo. The mutant viruses with individual C2441S, P2938S or R2985P signature substitutions, and with all three substitutions, showed significantly higher intracellular replication efficiencies and greater cytopathic effects than the parental JFH1 in vitro. The mutant HCV-inoculated mice showed significantly higher serum HCV RNA and higher level of expression of ER stress-related proteins in early period of infection. At 8 weeks post inoculation, these signature mutations had reverted to the wild type sequences. HCV-induced cytopathogenicity is associated with the level of intracellular viral replication and is determined by certain amino acid substitutions in HCV-NS5A and NS5B regions. The cytopathic HCV clones exhibit high replication competence in vivo but may be eliminated during the early stages of infection

HCV-JFH 1产生细胞病变斑的亚克隆（Sekine-Osajima Y，et al.，Virology 2008 ; 371：71）。在这里，我们研究了病毒的氨基酸取代细胞病变突变HCV-JFH 1克隆和它们的特点，在体外和体内。具有单独的C2441 S、P2938 S或R2985 P特征替换以及具有所有三种替换的突变体病毒在体外显示出比亲本JFH 1显著更高的细胞内复制效率和更大的细胞病变效应。在感染的早期，突变型HCV感染小鼠的血清HCVRNA和ER应激相关蛋白的表达水平显著升高。在接种后8周，这些特征突变已恢复为野生型序列。HCV诱导的细胞致病性与细胞内病毒复制水平相关，并由HCV-NS 5A和NS 5 B区域的某些氨基酸取代决定。细胞病变的HCV克隆在体内表现出高复制能力，但在感染的早期阶段可能被消除

项目成果

Inhibition of hepatitis C virus NS5B polymerase by S-trityl-L-cysteine derivatives.

• DOI: 10.1016/j.ejmech.2012.01.010
• 发表时间: 2012-03
• 期刊: European journal of medicinal chemistry
• 影响因子: 6.7
• 作者: Nichols DB;Fournet G;Gurukumar KR;Basu A;Lee JC;Sakamoto N;Kozielski F;Musmuca I;Joseph B;Ragno R;Kaushik-Basu N
• 通讯作者: Kaushik-Basu N

Association of gene expression involving innate immunity and genetic variation in IL28B with antiviral response

IL28B 中涉及先天免疫和遗传变异的基因表达与抗病毒反应的关联

• 发表时间: 2011
• 期刊: Hepatology
• 影响因子: 13.5
• 作者: Asahina Y;Sakamoto N;et al.
• 通讯作者: et al.

MMP-2 and MMP-14 derived from donor cells enhance therapeutic efficacy of liver cell transplantation in mice

供体细胞来源的 MMP-2 和 MMP-14 增强小鼠肝细胞移植的治疗效果

• 发表时间: 2011
• 作者: Kakinuma S;Sakamoto N;Watanabe M;et al.
• 通讯作者: et al.

IL-6-mediated intersubgenotypic variation of interferon sensitivity in hepatitis C virus genotype 2a/2b chimeric clones.

• DOI: 10.1016/j.virol.2010.07.041
• 发表时间: 2010-11
• 期刊: Virology
• 影响因子: 3.7
• 作者: G. Suda;N. Sakamoto;Yasuhiro Itsui;M. Nakagawa;Megumi Tasaka-Fujita;Yusuke Funaoka;Takako Watanabe;S. Nitta;Kei Kiyohashi;Seishin Azuma;S. Kakinuma;K. Tsuchiya;M. Imamura;N. Hiraga;K. Chayama;Mamoru Watanabe

Association between lipid accumulation and the cannabinoid system in Huh7 cells expressing HCV genes.

• DOI: 10.3892/ijmm.2011.622
• 发表时间: 2011-05
• 期刊: International journal of molecular medicine
• 影响因子: 5.4
• 作者: Mako Toyoda;A. Kitaoka;Kazuyuki Machida;T. Nishinakagawa;R. Yada;M. Kohjima;Masaki Kato;K. Kotoh