新型抗原亚群H9病毒的血凝素蛋白单克隆抗体的制备与鉴定  

Preparation and Identifi cation of Monolonal Antibodies Againist Hemagglutinin Protein of Novel Antigen Subgroup of H9 Avian Infl uenza Virus

作　　者：闫婉婉 李雪松[1] 宿鑫 石晓娜 陈新武 魏朋华 刘芹防[1] 杨健美[1] 闫大为[1] 张志飞 李泽君[1] 滕巧泱[1] YAN Wanwan;LI Xuesong;SU Xin;SHI Xiaona;CHEN Xinwu;WEI Penghua;LIU Qinfang;YANG Jianmei;YAN Dawei;ZHANG Zhifei;LI Zejun;TENG Qiaoyang(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》2023年第4期158-163,共6页Chinese Journal of Animal Infectious Diseases

基　　金：十三五重点研发计划(2018YFD0501506);上海自然科学基金(18ZR1449000)。

摘　　要：H9N2亚型禽流感病毒(AIV)抗原变异迅速,相继于2009年及2013年分别出现新的抗原亚群病毒,对该病毒的防控带来了极大的挑战。目前,关于H9N2禽流感病毒形成不同抗原亚群机制,尚不清楚。流感病毒血凝素蛋白(HA)的单克隆抗体是研究该机制的基础工具。为此,本研究通过血凝抑制(HI)实验筛选到1株新型的抗原亚群H9N2病毒H514。通过杂交瘤细胞融合技术以及间接免疫荧光的方法,筛选到5株抗H514病毒HA蛋白的单抗,均属于Ig G亚型。Western blot结果表明5株单克隆抗体中只有5E9作用的是线性表位,其余4株皆是针对构象表位。进一步地研究发现,这5株单克隆抗体对H514株皆有HI活性,最高达到2~8。这些结果为H9N2亚型禽流感病毒的抗原性分子研究提供了重要材料。The antigen of H9N2 subtype avian influenza virus(Avian influenza virus,AIV) has mutated rapidly,and different antigen subgroups appeared in 2009 and 2013,which brought great challenges to the prevention and control of this virus.What is the molecular basis of H9N2 AIV forming different antigen subgroups? Not clear so far.Monoclonal antibodies(MAbs) against Influenza virus hemagglutinin(HA) are the basic tools for studying the molecular mechanism.For this reason,we used the hemagglutination inhibition(HI) experiment and found a new antigenic subgroup of H9N2 virus H514 isolate.In this study,5 MAbs against the HA protein of the H514 virus were selected from hybridomas as screened by indirect immunofluorescence.All of these 5 MAbs belonged to the IgG subtype.Western blot showed that MAb 5E9 acted on the linear epitope of the HA protein and other 4 MAbs were all directed to the conformational epitopes.Further research found that all 5 MAbs had HI activity against H514 virus with titer up to 2~8.These results provided important materials for studying molecular biology of H9N2 subtype AIV.

关 键 词：H9N2亚型 禽流感病毒 抗原变异 单克隆抗体 血凝素 

分 类 号：S852.659.5[农业科学—基础兽医学]