传染性支气管炎病毒诱导Vero细胞发生自噬促进病毒的复制  

IBV promotes self-replication by inducing autophagy in Vero cells

作　　者：杨雪晴 孙军峰 李慧昕[2] 韩宗玺[2] 刘胜旺[2] YANG Xueqing;SUN Junfeng;LI Huixin;HAN Zongxi;LIU Shengwang(College of Animal Science and Technology,Heilongjiang Bayi A gricultural University,Daqing 163319,China;Division of Avian Infectious Diseases,State Key Laboratory for Animal Disease Control and Prevention,Harbin Veterinary Research Institute,Chinese Academy of A gricultural Sciences,Harbin 150069,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]中国农业科学院哈尔滨兽医研究所,动物疫病防控全国重点实验室,禽呼吸道传染病创新团队,黑龙江哈尔滨150069

出　　处：《中国兽医科学》2024年第8期1019-1026,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2021YFD1801105);黑龙江省自然科学基金团队项目(TD2021C001);国家蛋鸡产业体系技术岗位科学家项目(CARS-40)。

摘　　要：为探究禽传染性支气管炎病毒(infectious bronchitis virus,IBV)能否诱导细胞自噬及其对病毒复制的影响,本研究利用IBV细胞适应株CEA接种DF-1和Vero细胞,通过Western-blot、透射电镜和激光共聚焦显微镜检测自噬通路关键蛋白LC3和P62的表达和分布以及自噬小体的形成情况,并通过自噬抑制剂和诱导剂处理评估了自噬对IBV复制的影响。结果显示,IBV不能在DF-1细胞中诱导LC3的转化,但能够诱导Vero细胞中LC3-II和P62的水平升高,并能在Vero细胞中诱导形成自噬小体以及LC3-II向自噬体中的聚集,表明IBV能够诱导Vero细胞发生完整自噬。用自噬抑制剂渥曼青霉素和氯喹处理细胞能够抑制IBV诱导的自噬以及病毒的复制,而用自噬激活剂雷帕霉素处理则能够促进病毒的复制。综上所述,IBV能够在Vero细胞中诱导自噬的发生,从而促进病毒的复制。In this study,to investigate whether avian Infectious bronchitis virus(IBV)can induce autophagy in cells and its effect on viral replication,DF-1 and Vero cells were inoculated with cell-adapted strain CEA of IBV.Then the expression and distribution of key proteins of autophagy pathway,LC3 and P62,and the formation of autophagosomes were detected by Western-blot,transmission electron microscopy and confocal laser microscopy,respectively.The effect of autophagy on IBV replication was further evaluated by the treatment of autophagy inhibitor and inducer.The results showed that IBV did not induce LC3 transformation in DF-1 cells,but could increase levels of LC3-II and P62 in Vero cells,as well as the formation of autophagosomes in Vero cells and the aggregation of LC3-II into autophagosomes,suggesting that IBV could induce complete autophagy in Vero cells.The treatment with autophagy inhibitors,chloroquine and Wortmannin,inhibited IBV-induced autophagy and viral replication in Vero cells,meanwhile the treatment with autophagy activator rapamycin promoted viral replication.In summary,IBV could induce autophagy in Vero cells and promote viral replication.

关 键 词：禽传染性支气管炎病毒 自噬 病毒复制 

分 类 号：S852.659.6[农业科学—基础兽医学]