摘要
目的快捷筛选到适合丙型肝炎病毒（HCV）培养的细胞系。方法将荧光素酶报告基因耦联到JFHlHCV基因组中，检测了多个人肝细胞系对HCV的易感性，进一步利用耦联荧光素酶的HCV复制子模型筛选并经过IFN-Ot处理更适宜HCV培养的Huh7亚细胞克隆株。结果实验结果显示BEL7402、BEL7404、QSG7701、SMMC7721、QGY7701、QGY7703和HepG等肝细胞均不易感染HCV，仅限于Huh7细胞能被HCV感染。同时实验获得了更适宜HCV培养的Huh7亚细胞克隆株——Huh7G3。结论在利用IFN-仅处理细胞的过程中，通过检测荧光素酶活性，显示IFN-Ot对HCV亚基因组的复制抑制效果具有剂量依赖效应，研究同时显示：HCV复制子细胞能否抵抗HCV的超感染依赖于细胞内是否存在HCVRNA复制而不是其复制水平的高低。
Objective To select appropriate cell line for Hepatitis C virus （HCV） production. Methods A JFHl-based luciferase reporter virus HCV-tGlu was generated. Using the reporter virus, sever- al human hepatic cell lines were checked for their permissiveness to HCV infection. Furthermore, HCV bicistronic reporter system for subgenomic replication and IFN-α treatment approach were used to establish a new Huh7 subclone cell line. Results The results showed that HCV infection was restricted to Huh7, but not to BELT402, BELT404, QSG7701, SMMC7721, QGY7701, QGY7703 or HepaG2 cell lines. An IFN-a treated Huh7 subclone cell line, Huh7G3, which bear an increased permissiveness for HCVcc replication was established. Conclusion During IFN treatment, the inhibitory effect of IFN on HCV subgenome replication was dose-dependent as indicated by the luciferase activities. The results also showed that the resistance of HCV replicon-containing cells to the HCV superinfection relied on the presence of HCV RNA in the ceUs but not denend on the intracellular virus zenomic reolication level.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2013年第3期161-167,共7页
Chinese Journal of Microbiology and Immunology
基金
国家科技重大专项重大新药创制课题（2012ZX09105302）
中国医学科学院医学生物学研究所人才引进科研启动费项目
病原生物学研究所基本科研业务费项（2011IPBl07）
协和新星专项资金资助
中央高校基本科研业务费专项资金资助（2012Y08）
关键词
丙型肝炎病毒
细胞培养
报告基因
超感染
Hepatitis C virus
Cell culture
Reporter gene
Super infection
