抗H9亚型禽流感病毒血凝素广谱单克隆抗体的筛选与鉴定  

CHARACTERIZATION OF BROAD-SPECTRUM MONOCLONAL ANTIBODIES AGAINST HEMAGGLUTININ OF H9 SUBTYPE AVIAN INFLUENZA VIRUS

作　　者：屈孟锦 李雪松[1] 滕巧泱[1] 李鲁兆 刘性坡 崔宏锐 杨健美[1] 李泽君[1] QU Meng-jin;LI Xue-song;TENG Qiao-yang;LI Lu-zhao;LIU Xing-po;CUI Hong-rui;YANG Jian-mei;LI Ze-jun(Shanghai Veterinary Research Institute,CAAS,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：《中国动物传染病学报》2020年第6期48-53,共6页Chinese Journal of Animal Infectious Diseases

基　　金：国家重点研发计划(2016YFD0500800,2017YFD0500806);国家自然科学基金项目(31702237,31771000);上海市自然科学基金项目(17ZR1437400);上海科委创新行动计划农业项目(17391901700)

摘　　要：本研究通过血凝抑制实验和间接免疫荧光实验,在前期利用表达H9N2亚型禽流感病毒(AIV)A/Chicken/Shanghai/441/2009(H9N2)(SH441)血凝素(HA)基因的真核表达质粒免疫小鼠,成功获得多株抗H9亚型HA蛋白的单克隆抗体中,筛选到4株与2009-2015年间分离到的H9N2 AIV具有良好反应性的单克隆抗体;中和实验表明,这4株单抗均对SH441毒株具有中和活性;通过细胞瞬时表达HA蛋白和间接免疫荧光实验发现,这4株单抗均可以与HA的HA1区域结合,不与HA2反应。以上结果表明,筛选获得的4株单抗均可识别H9N2 AIV HA1上相对保守的氨基酸序列,对H9N2 AIV具有广谱的血凝抑制活性,将为H9N2 AIV通用疫苗研制及高通量抗体检测方法的建立提供重要材料。In the previous study,multiple monoclonal antibodies(MAbs)against H9 subtype was obtained by immunizing mice with eukaryotic expression plasmid expressing H9N2 subtype Avian influenza virus(AIV)A/Chicken/Shanghai/441/2009(H9N2)(SH441)hemagglutinin(HA)gene.In this study,four MAbs with good reactivity with H9N2 AIV isolated from 2009 to 2015 were screened by hemagglutination inhibition(HI)test and indirect immunofluorescence assay(IFA).The results showed that these 5 MAbs had neutralizing activity against SH441 strain.These four MAbs bound to the HA1 region of HA and did not react with HA2 by transient expression of HA protein with cells and IFA.In conclusion,these four MAbs recognized the relatively conserved amino acid sequence on H9N2 AIV HA1 and had broad-spectrum HI activity on H9N2 AIV,which would provide important materials for the development of H9N2 AIV universal vaccine and high-throughput antibody detection method.

关 键 词：H9N2亚型禽流感病毒 血凝素 单克隆抗体 广谱性 

分 类 号：S852.659.5[农业科学—基础兽医学]