细胞自噬对甲型流感病毒H1 N1感染A549细胞过程中病毒复制及Ⅰ型干扰素表达的影响  

Effects of autophagy on viral replication and typeⅠ interferon expression in A549 cells during influ-enza A virus （H1N1） infection

作　　者：李嘉祺[1] 胡雅洁[1] 宋杰[1] 李洪哲[1] 王晶晶[1] 郭磊[1] 郑惠文[1] 宁若彤 范海涛 杨泽宁[1] 刘龙丁[1] Li Jiaqi Hu Yafie Song Jie Li Hongzhe Wang Jingfing Guo Lei Zheng Huiwen Ning Ruotong Fan Haitao Yang Zening Liu Longding(Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Disease, Institute of Medical Biology, Chinese Academy of Medical Sciences ＆ Peking Union Medical College, Kunming 650118, China)

机构地区：[1]中国医学科学院北京协和医学院,医学生物学研究所,云南省重大传染病疫苗研发重点实验室,昆明650118

出　　处：《中华微生物学和免疫学杂志》2017年第2期85-90,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（31300143）;云南省应用基础研究计划（2015FB193）

摘　　要：目的 通过研究细胞自噬在甲型流感病毒H1 N1感染细胞过程中对病毒复制及Ⅰ型干扰素表达的影响,为进一步阐明流感病毒感染机制提供实验资料.方法 利用自噬抑制剂三甲基腺嘌呤（3-methyladenine,3-MA）抑制细胞自噬,流式细胞术检测3-MA对细胞凋亡的影响,激光共聚焦显微镜观察H1N1感染A549细胞后的自噬现象,利用real-time PCR检测自噬被抑制前后病毒拷贝数的变化以及Ⅰ型干扰素（typeⅠinterferon,IFN-Ⅰ）产生通路各基因mRNA表达量的变化.分析细胞自噬对病毒复制及干扰素表达的影响.结果 当H1N1感染A549细胞后可诱导细胞产生不完全自噬,导致自噬体积累.当自噬被3-MA抑制后,TLR3-TLR7-MyD88-IRF7-IFN-α/β信号通路各基因mRNA表达量上调,与此同时病毒拷贝数及血凝滴度下调.结论 H1N1可能通过诱导不完全自噬逃逸了机体的抗病毒固有免疫应答,促进了自身复制和存活.Objective To investigate the effects of autophagy on viral replication and type Ⅰ in-terferon （IFN-Ⅰ） expression in A549 cells during influenza A virus （H1N1） infection for providing refer-ences for further elucidation of the mechanisms of influenza virus infection. Methods The autophagy inhibi-tor 3-methyladenine （3-MA） was used to inhibit autophagy. Flow cytometry was used to detect the effects of 3-MA on cell apoptosis. Status of autophagy after infecting A549 cells with H1N1 virus was observed under laser confocal microscopy. Real-time PCR was used to detect the changes in viral copy number and in the mRNA levels of various genes related to the IFN-Ⅰ signaling pathway before and after inhibition of autoph-agy. Results Infecting A549 cells with H1N1 virus induced incomplete autophagy and resulted in autopha-gosome accumulation. Inhibiting the autophagy of A549 cells with 3-MA up-regulated the expression of vari-ous genes in TLR3-TLR7-MyD88-IRF7-IFN-α/β signaling pathway at mRNA level, but down-regulated the viral copy number and hemagglutinin （HA） titer. Conclusion Influenza A virus （H1N1） might be able to facilitate its replication and survival by inducing incomplete autophagy to escape from the innate immune re-sponses.

关 键 词：细胞自噬 甲型流感病毒 A549细胞 Ⅰ型干扰素 

分 类 号：R373[医药卫生—病原生物学]