摘要
目的:探究Ku70在cGAS介导的抗DNA病毒天然免疫反应中的调控作用。方法:蛋白免疫沉淀实验检测Ku70与cGAS相互作用。荧光素酶报告基因实验验证Ku70是否调控cGAS信号通路。采用特异shRNA沉默THP-1细胞中靶基因Ku70表达,qPCR和Western blot检测下游因子表达。结果:Ku70与cGAS存在相互作用。过表达Ku70显著增强HEK293T细胞中cGAS介导的IFN-β启动子活性。干扰Ku70表达显著降低DNA病毒诱导的Ⅰ型干扰素mRNA和蛋白表达。结论:Ku70参与cGAS介导的天然免疫反应,并在DNA病毒刺激下,上调Ⅰ型干扰素信号通路活性,进而正向调控宿主抗病毒的天然免疫信号反应。
Objective:To investigate regulatory role of Ku70 in cGAS-mediated innate immune responses against DNA viruses.Methods:Co-immunoprecipitation experiments to detect interaction between Ku70 and cGAS.Luciferase reporter gene experiment to verify whether Ku70 regulated cGAS-mediated signaling pathway.Ku70 expression in THP-Ⅰwas silenced with specific shRNA,and expressions of downstream genes were detected by qPCR and Western blot.Results:There was an interaction between Ku70 and cGAS.Overexpression of Ku70 dose-dependently enhanced cGAS-mediated IFN-βactivity in HEK293T cells.Ku70 silencing significantly reduced expression of typeⅠinterferon induced by DNA viruses.Conclusion:Ku70 was involved in cGAS-mediated innate immune response.Upon DNA virus infection,Ku70 upregulated production of typeⅠinterferon,and therefore positively regulated host antiviral innate immune responses.
作者
储贝
孙钦秒(指导)
CHU Bei;SUN Qin-Miao(School of Life Sciences,University of Science and Technology of China,Hefei 230027,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第18期2177-2181,共5页
Chinese Journal of Immunology
基金
国家自然科学基金项目(31970895)。