Sn受体N端V-set结构域和CD163受体SRCR5结构域在PRRSV-ADE感染中的作用  

Roles of N-terminal V-set domain of Sn receptor and SRCR5 domain of CD163 receptor in PRRSV-ADE infection

作　　者：张留君 师瑞龙 王焕弟 赵磊[1] 王大胜 叶春妹 郭敏娜 刘德义[1] 夏平安[2] ZHANG Liujun;SHI Ruilong;WANG Huandi;ZHAO Lei;WANG Dasheng;YE Chunmei;GUO Minna;LIU Deyi;XIA Pingan(College of Animal Science,Anhui Science and Technology University,Fengyang,Anhui 233100,China;College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]安徽科技学院动物科学学院,安徽凤阳233100 [2]河南农业大学动物医学院,河南郑州450046

出　　处：《中国兽医学报》2022年第12期2458-2463,共6页Chinese Journal of Veterinary Science

基　　金：安徽省高等学校自然科学研究重点资助项目(KJ2021A0870);安徽科技学院大学生创新创业训练计划创新训练资助项目(省级)(S202110879173)。

摘　　要：为了研究Sn受体N端V-set结构域和CD163受体SRCR5结构域在猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)抗体依赖性增强作用(ADE)感染中的作用,对2种结构域蛋白进行表达、纯化,并免疫大白兔制备其特异性多克隆抗体IgG。应用制备的抗Sn-N-V-set IgG、CD163-SRCR5 IgG和正常兔IgG分别封闭PAMs细胞,然后使用猪抗PRRSV IgG与PRRSV悬液混合后制成的感染性免疫复合物感染已封闭的PAMs细胞。感染后每隔12 h收集细胞上清液1次,用已建立的实时荧光定量RT-PCR方法测定PRRSV的RNA拷贝数,同时以Marc-145细胞测定PRRSV的病毒滴度。结果显示,当Sn受体N端V-set结构域和CD163受体SRCR5结构域被其特异性抗体封闭后,能够显著抑制体外PRRSV感染PAMs细胞的抗体依赖性增强作用(ADE)(P<0.01)。结果表明,2种受体结构域均参与PRRSV-ADE感染。To study the roles of Sn receptor N-terminal V-set domain and CD163 receptor SRCR5 domain in PRRSV-ADE infection,the domain proteins of Sn receptor N-terminal V-set and CD163 receptor SRCR5 were expressed,purified and used to generate the anti-Sn-N-V-set IgG and anti-CD163-SRCR5 IgG by immunizing the white rabbits.PAMs cells were pre-blocked with anti-Sn-N-V-set IgG,anti-CD163-SRCR5 IgG or rabbit-negative IgG before they were infected with PRRSV+ICs.The cell culture supernatants collected every 12 h post-infection were used to measure the PRRSV RNA copies and its virus titers by the established real-time fluorescence quantitative RT-PCR and the Reed-Muench method,respectively.The results showed that the blocking of Sn receptor N-terminal V-set domain or CD163 receptor SRCR5 domain significantly inhibited PRRSV-ADE infection in PAMs in vitro(P<0.01),suggesting that the N-terminal V-set domain of Sn receptor and the SRCR5 domain of CD163 receptor were involved in PRRSV-ADE infection.

关 键 词：Sn受体 CD163受体 猪繁殖与呼吸综合征病毒(PRRSV) 抗体依赖性增强作用(ADE) 

分 类 号：S852.2[农业科学—基础兽医学] S852.65[农业科学—兽医学]