摘要
非洲猪瘟(African swine fever,ASF)是由非洲猪瘟病毒(African swine fever virus,ASFV)感染引起的高致死率传染病。骨髓基质细胞抗原2(BST-2,也称为Tetherin/HM1.24/CD317)被鉴定为一种抑制包膜病毒释放的新型宿主限制因子,它可以将病毒颗粒结合到细胞表面,随后在溶酶体中进行内化和降解,从而抑制病毒复制。因此,它对宿主的抗病毒作用十分重要。本实验室前期研究表明,怀孕猪血清会促进ASFV的复制。ITRAQ分析表明,BST-2蛋白属于下调前十的蛋白之一。为探究宿主蛋白BST-2与ASFV复制之间的关系,本试验通过蛋白免疫印迹(Western-blot)和实时荧光定量PCR(RT-qPCR)分析了ASFV感染对BST-2表达的影响;构建并合成了BST-2真核表达质粒以及siRNA干扰序列,通过RT-qPCR、Western-blot去探究外源过表达BST-2及敲低BST-2对ASFV体外复制的调控作用。结果显示,外源过表达BST-2显著抑制了ASFV的复制,敲低BST-2促进了ASFV的复制。本研究通过对BST-2进行过表达和敲低,证实了BST-2作为一种宿主抗病毒分子能抑制ASFV的体外复制,为进一步深入研究宿主蛋白抗ASFV功能提供了新的途径,也为ASF的防控提供了新思路。
African swine fever(ASF)is a highly lethal infectious disease caused by the African swine fever virus(ASFV).Bone marrow stromal cell antigen 2(BST-2,also known as Tetherin/HM1.24/CD317),which binding virus particles to the cell surface,internalizing them,and subsequently degrading them in lysosomes,has been identified as a novel host factor that restricts the release of enveloped virus particles.That’s why BST-2 can effectively inhibits virus replication.Moreover,BST-2 plays a crucial role in the host's antiviral effect.Notably,previous research conducted in our laboratory has demonstrated that pregnant pig serum can enhance the replication of ASFV.Present study utilized ITRAQ to deanalyzethe disparity in proteins between empty pregnant pig serum and pregnant pig serum,revealingthat BST-2 protein was one of the top ten down-regulated proteins.In order to investigate the relationof host protein BST-2 and ASFV,the effect of ASFV infection on BST-2 expression was assessed throughWestern-blot and real-time fluorescence quantitative PCR(RT-qPCR).Furthermore,we constructed andsynthesized the eukaryotic expression plasmid of BST-2 and siRNA interference sequence.Using RT-qPCRand Western - blot analysised the regulation function of exogenous overexpression and knock-down ofBST-2 on the in vitro replication of ASFV.The findings revealed a significant inhibition of ASFV replicationupon exogenous overexpression of BST-2,while knock-down of BST-2 facilitated ASFV replication.This article demonstrated the overexpression and knockdown of BST-2,establishing its role as a hostantiviral molecule that effectively impedes the replication of ASFV in vitro.These findings offer anovel avenue for future investigations into the antiviral function of the host proteins against ASFV,while also providing a new idea for the prevention and control of ASF.
作者
赵美玉
杨行
史喜绢
张大俊
赵登率
陈玲玲
别鑫恬
闫文倩
陈国辉
赵思越
李平
郑海学
栗孟飞
张克山
ZHAO Meiyu;YANG Xing;SHI Xijuan;ZHANG Dajun;ZHAO Dengshuai;CHEN Lingling;BIE Xintian;YAN Wenqian;CHEN Guohui;ZHAO Siyue;LI Ping;ZHENG Haixue;LI Mengfei;ZHANG Keshan(College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;State Key Laboratory for Animal Disease Control and Prevention/College of Veterinary Medicine of Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730000,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2024年第2期143-150,共8页
Chinese Veterinary Science
基金
国家重点研发计划项目(2021YFD1801300)
国家生猪产业体系(CARS-35)
中央高校基本科研业务费专项
国家生猪技术创新中心先导科技项目(NCTIP-XD/C03)
甘肃省科技重大专项(22ZD6NA001)
“十四五”广东省农业科技创新十大主攻方向“揭榜挂帅”项目(2022SDZG02)
中国农业科学院科技创新工程(CAAS-ASTIP-2022-LVRI)
中国农业科学院基础科学中心重大任务(CAAS-CSLPDC-202302)。
