利用丙型肝炎病毒复制子模型检测重组人λ2干扰素的体外活性

Detection of Interferon λ2 Activity In Vitro by HCV Replicon Model

目的:利用丙型肝炎病毒(HCV)复制子细胞模型,体外研究重组人λ2干扰素(hIFNλ2)对HCV复制子的抑制作用。方法:构建重组质粒pcDNA3.1-hIFNλ2-HA,转染293T细胞,表达带有HA标签的hIFNλ2,表达后的上清通过Western印迹检测,然后稀释至不同浓度后作用于带有HCV复制子的Huh7.5细胞系,每隔24h取样并更换培养液,通过报告基因检测hIFNλ2对HCV复制子的抑制作用。结果:构建了pcDNA3.1-hIFNλ2-HA真核表达重组体,并在293T细胞中表达出hIFNλ2,该蛋白能有效抑制HCV复制子的复制。结论:hIFNλ2对HCV复制子具有抑制作用。

Objective:To study the inhibitory effect of recombinant human interferonλ2(hIFNλ2)on HCV replicon in vitro by using cell culture system of HCV replicon.Methods:This study constructed recombinant plasmidpcDNA3.1-hIFNλ2-HA,and expressed the hIFNλ2with HA tag in293T cells,then detected the supernatant ofthe cell by Western blot,and then used supernatant diluted with different ratio to effect on Huh7.5cell culturesystem with HCV replicon,samples were collected every24hours and replaced the medium,finally using the report gene to detect the inhibitory effect of the hIFNλ2in HCV virus replicon.Results:The eukaryotic expressionrecombinant plasmid pcDNA3.1-hIFNλ2was successfully contructed,and hIFNλ2was expressed in293T cells,and confirmed that the hIFNλ2could inhibit the replication of HCV.Conclusion:The results proved that hIFNλ2has a direct inhibitory effect on HCV replicon.