蓝舌病病毒感染对BHK-21细胞中Ⅰ型干扰素应答的影响  

Effect of bluetongue virusinfection on typeⅠinterferon response in BHK-21 cells

作　　者：罗世美 陈韵伊 李其沙 周艳梅 王怡斐 廖芯玉 胡雪柔 魏远健 李梦琴 朱萌 张迅 陈贝蕊 马鲜平 谢佳芮 寇美玲 苗海生[2] 李芳 易华山 LUO Shimei;CHEN Yunyi;LI Qisha;ZHOU Yanmei;WANG Yifei;LIAO Xinyu;HU Xuerou;WEI Yuanjian;LI Mengqin;ZHU Meng;ZHANG Xun;CHEN Beirui;MA Xianping;XIE Jiarui;KOU Meiling;MIAO Haisheng;LI Fang;YI Huashan(College of Veterinary Medicine,Southwest University,Chongqing 402460,China;Yunnan Tropical and Subtropical Animal Virus Disease Laboratory,Yunnan Veterinary and Animal Science Institute,Kunming 650224,China;Dazu Center for Animal Disease Prevention and Control,Dazu,Chongqing 402360,China)

机构地区：[1]西南大学动物医学院,重庆荣昌402460 [2]云南省畜牧兽医科学院云南省热带亚热带动物病毒病重点实验室,云南昆明650224 [3]重庆市大足区动物疫病预防控制中心,重庆大足402360

出　　处：《中国兽医学报》2024年第8期1639-1644,1690,共7页Chinese Journal of Veterinary Science

基　　金：重庆市自然科学基金面上资助项目(CSTB2022NSCQ-MSX0419);重庆市基础研究与前沿探索专项资助项目(cstc2018jcyjAX0615)。

摘　　要：蓝舌病病毒(bluetongue virus,BTV)是一种严重危害绵羊等反刍动物的一种虫媒病毒,为研究蓝舌病病毒感染与宿主细胞干扰素抗病毒免疫应答的分子机制。研究以感染复数(MOI)为1的BTV诱导18、24、36 h的BHK-21细胞通过实时荧光定量PCR分析干扰素通路基因mRNA表达特征,以及Western blot分析MDA5、TRAF3、RIG-Ⅰ和TBK1蛋白表达。结果表明,在诱导24 h时,干扰素信号通路基因表达的变化最为明显,IFN-α、IFN-β、RIG-Ⅰ、TBK1、MDA5、VISA、TRAF3基因的mRNA表达水平呈上调表达,而IKKε、TRAF6基因的mRNA表达水平呈下调表达,在蛋白水平上MDA5、TBK1蛋白上调表达而RIG-Ⅰ、TRAF3蛋白下调表达,表明BTV感染诱导BHK-21细胞中Ⅰ型干扰素免疫应答,为进一步探究IFN-Ⅰ信号通路调控基因在BTV感染的宿主细胞抗病毒免疫机制研究奠定基础。Bluetongue virus is an arbovirus that seriously harms ruminants such as sheep,this study aims to investigate the molecular mechanism of bluetongue virus infection and host cell interferon antiviral immune response.The study was conducted to characterize the mRNA expression of interferon pathway genes by real-time fluorescence quantitative PCR,as well as Western blot analysis of MDA5,TRAF3,RIG-Ⅰ,and TBK1 protein expression in BHK-21 cells induced by BTV with a multiplicity of infections(MOI)of 1 for 18,24,and 36 h.The results showed that the most pronounced changes in the expression of interferon signaling pathway genes were observed at 24 h of induction,the gene mRNA expression levels of the IFN-α,IFN-β,RIG-Ⅰ,TBK1,MDA5,VISA,and TRAF3 genes were upregulated.However,the mRNA expression levels of IKKεand TRAF6 genes were downregulated.At the protein level,MDA5 and TBK1 proteins were upregulated while RIG-I and TRAF3 proteins were downregulated,which showed that BTV infection induces a typeⅠinterferon immune response in BHK-21 cells.This study lays the foundation for further exploring the antiviral immunity mechanism of IFN-Ⅰsignaling pathway regulatory genes in host cells infected with BTV infection.

关 键 词：蓝舌病病毒 天然免疫 Ⅰ型干扰素 BHK-21 

分 类 号：S852.65[农业科学—基础兽医学]