欧洲禽源NA和M基因对重组猪流感病毒致病性的影响  

Impact of European avian-like NA and M genes on pathogenicity of reassortant swine influenza viruses

作　　者：刘晓敏[1] 宫晓倩[1] 阮宝阳 张鹏[1] 汪琪[1] 杨海明[1] 单同领[1] 童武[1] 李国新[1] 郑浩[1] 周艳君[1] 童光志[1] 于海[1] LIU Xiao-min GONG Xiao-qian RUAN Bao-yang ZHANG Peng WANG Qi YANG Hai-ming SHAN Tong-ling TONG Wu LI Guo-xin ZHENG Hao ZHOU Yan-jun TONG Guang-zhi YU Hai(Shanghai Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Shanghai 200241, China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国兽医科学》2016年第11期1341-1348,共8页Chinese Veterinary Science

基　　金：上海市自然科学基金青年项目(16ZR1444000;12ZR1453500);国家青年自然科学基金项目(31201916);中央级公益性科研院所基本科研业务费项目(2015JB07);中国农业科学院创新工程"猪病毒性繁殖障碍综合征团队"项目

摘　　要：为探究欧洲禽源N A和M基因对重组猪流感病毒致病性的影响,通过对猪流感病毒A/swine/Shanghai/1/07(H 1N2)(S12)与A/swine/Zhejiang/1/07(H1N1)(Z11)进行测序以及基因型分析,确定了2009甲型H1N1流感病毒的PB 2、PB1、PA、HA、NP和NS基因均起源于三源重组H1N2亚型SIV(S12),NA和M基因均起源于欧洲禽源H1N1亚型SIV(Z11)。利用RT-PCR技术对Z11的NA和M基因分别进行扩增及克隆,以S12为骨架,成功拯救到含有欧洲禽源NA、M基因的重组病毒。通过检测不同病毒在MD CK、A549、PK15细胞上的生长特性,并以小鼠为模型对不同病毒的致病性进行了差异性分析。研究结果表明,欧洲禽源H1N1亚型SIV的NA和M基因相互协同地提高了病毒在体外多种细胞以及小鼠肺内的复制能力,增强了病毒对小鼠的致病性,这为甲型H1N1流感病毒的分子致病机制研究提供了理论依据,对猪流感的防控及预警具有重要意义。To determine the pathogenicity of reassortant swine influenza viruses containing European avian-like NA or M gene in mammals,A/swine/Shanghai/i/07 （H1N2） （S12）and A/swine/Zhejiang/1/07 （H1N1） （Zll）isolated by our laboratory were sequenced. By genetic analysis of PB2,PB1,PA,HA,NPand NS genes showed that 2009 pandemic H1N1 influenza A virus originated from triple-reassortant H1N2 swine influenza virus（S12）,the NA and M genes were closely related to European avian-like H1N1 swine influenzavirus（Zll）.The NA and M genes of Zll were amplified by RT-PCR,separately, and cloned into the vector. Basedon the reverse genetic system of S12,reassortant viruses bearing NAor M gene from Zll were rescued successfully. To evaluate the replicative ability of reassortant swine influenza viruses, the growth kinetics were measured in MDCK,A549 and PK-15 cells. To investigate the impact of European avian-likedNAor M gene on pathogenicity,we compared the virulence of different viruses using a BALB/cmouse model. The results show that the NAand M genes of European avian-like H1N1 swine influenza virus synergistically enhance the ability of replication both in vitro and in vivo. Besides,the combination of European avian-likeNAand M genes significantly increase the virulence in mice. Our findings provide theoretical foundation for the molecular pathogenesis of pandemic H1N1 influenza A virus,which are of great significance for the control and the early warning of the swine influenza.

关 键 词：欧洲禽源H1N1亚型SIV 基因型 重组病毒 生长特性 致病性 

分 类 号：S852.612[农业科学—基础兽医学]