蓝舌病病毒NS3蛋白抑制细胞Ⅰ型干扰素的产生  

Bluetongue virus NS3 protein inhibiting the production of type Ⅰ interferon from epithelial cells

作　　者：朱沛 肖雷[1] 吴梅 李珂 信爱国[1] ZHU Pei;XIAO Lei;WU Mei;LI Ke;XIN Aiguo(Yunnan Tropical and Subtropical Animal Virus Disease Laboratory,Yunnan Animal Science and Veterinary Institute,Kunming 650224,China)

机构地区：[1]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

出　　处：《中国兽医学报》2022年第6期1156-1162,1204,共8页Chinese Journal of Veterinary Science

基　　金：云南省畜牧兽医科学院自主研究资助项目(2022RW008);云南省应用基础研究计划青年资助项目(2017FD056)。

摘　　要：蓝舌病病毒(bluetongue virus, BTV)是一种双链RNA病毒,能感染多种细胞诱导强烈的Ⅰ型IFN效应,而BTV非结构蛋白NS3对IFN-β的生成有强烈抑制作用,但目前BTV感染与诱导IFN-Ⅰ产生的关联仍不清楚,BTV NS3蛋白如何影响IFN-Ⅰ合成也有待研究。为此本研究通过BTV-1和BTV-16感染人非小细胞肺癌细胞(A549)和犊牛原代肾细胞(MDBK)探索不同血清型、不同病毒载量、不同作用时间的BTV对细胞IFN-Ⅰ产生的影响,通过构建BTV NS3真核表达质粒,探索NS3蛋白对细胞IFN-Ⅰ转录及表达水平的调控和影响。qPCR结果表明BTV-1和BTV-16能感染A549和MDBK并诱发强烈的IFN-β效应,双荧光素酶报告基因结果显示BTV NS3对BTV诱导的IFN-β启动子活性具有抑制作用,且表达量与抑制作用呈正相关。本研究揭示了BTV增殖与细胞产生IFN-Ⅰ的关系,阐明了BTV NS3蛋白对IFN-β转录和表达水平的影响,其结果有助于了解BTV的致病机理及其干扰宿主的天然免疫应答机制,对BTV疫苗研发及抗病毒治疗具有科学意义。Bluetongue virus(BTV) is double-stranded RNA virus which infected multiple cells and induce strong IFN-Ⅰ effect,while BTV NS3 protein can strongly inhibit the synthesis of IFN-β,meanwhile the link between BTV infection and IFN-Ⅰ expression is still not clear,and how NS3 protein affects the signaling pathway of IFN-β synthesis remains to be studied.To clarify the connection between BTV replication efficiency and IFN-Ⅰ production,and the impact of IFN-Ⅰ when over expression of NS3 protein in cell,the study used different BTV serotype,different infection quantity and different infection time to explore the impact on the IFN-Ⅰ production in human nonsmall cell lung cancer cells(A549) and calf primary kidney cells(MDBK).BTV nonstructural protein NS3 eukaryotic expression plasmid was constructed to explore the influence of NS3 protein on IFN-Ⅰ transcription regulation and expression level.The results showed that both BTV-1 and BTV-16 could infect A549 and MDBK and induce strong IFN-β response,the overexpression of NS3 protein in cell weakly stimulated the production of IFN-β mRNA,while the dual luciferase report gene showed that NS3 protein could obviously induce the stimulation of BTV on IFN-β promoter.The research revealed the influence on the induction of IFN-Ⅰ by BTV-1 and BTV-16,illuminated the effect of NS3 protein on IFN-β transcription and expression level,these results help to understand pathogenic mechanism of BTV and the mechanism of interfering with the natural immune response of the host,as well as whether NS3 protein affect the production of IFN-Ⅰ,which has scientific significance on BTV vaccine development and antiviral therapy.

关 键 词：蓝舌病病毒 NS3蛋白 病毒感染 天然免疫 Ⅰ型干扰素 

分 类 号：S852.65[农业科学—基础兽医学]