蝙蝠流感病毒与H9N2亚型禽流感病毒M蛋白的免疫原性比较研究  

COMPARATIVE STUDY ON IMMUNOGENICITY FOR M PROTEINS OF BAT INFLUENZA VIRUS AND H9N2 AVIAN INFLUENZA VIRUS

作　　者：黄敏 刘宸瑞 滕巧泱[2] 刘芹防[2] 李雪松[2] 陈鸿军[2] 车广胜 任超超[2] 崔宏锐 闫大为[2] 周伟光[1] 李泽君[2,3] 杨健美 HUANG Min;LIU Chen-rui;TENG Qiao-yang;LIU Qin-fang;Li Xue-song;CHEN Hong-jun;CHE Guang-sheng;REN Chao-chao;CUI Hong-rui;YAN Da-wei;ZHOU Wei-guang;LI Ze-jun;YANG Jian-mei(Veterinary School of Inner Mongolia Agricultural University, Hohhot 010018, China;Shanghai Veterinary Research Institue, CAAS, Shanghai 200241, China;Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 200240, China)

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018 [2]中国农业科学院上海兽医研究所,上海200241 [3]上海市兽医生物技术重点实验室,上海200240

出　　处：《中国动物传染病学报》2018年第3期7-15,共9页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金项目(青年科学基金项目31700136);上海市兽医生物技术重点实验室开放基金项目(klab201706)

摘　　要：为比较新发蝙蝠流感病毒H17N10亚型与禽流感病毒H9N2亚型的M蛋白(包括M1和M2蛋白)的免疫原性,分别构建两种亚型流感病毒M1、M2的原核表达载体和真核表达载体,利用前期反向遗传学技术拯救获得两种重组病毒感染MDCK细胞,并分别与相应的单抗或多抗进行免疫印迹和免疫荧光鉴定。结果显示,两种亚型病毒的M1蛋白的原核表达产物均与M1多抗反应,均不与2株M1单抗反应;拯救的两种重组病毒感染细胞后及其M1蛋白的真核表达产物均与M1的单抗5F2反应,不与单抗3G8及M1的多抗反应,两种亚型的结果一致;而两种亚型的M2蛋白通过与单抗和多抗反应,出现了不一致的结果,M2的单抗仅与蝙蝠流感病毒M2的原核表达产物反应,不与H9N2禽流感病毒M2的原核表达产物反应,M2的多抗则反之;且M2的多抗仅与H9N2禽流感M2的真核表达产物以及拯救病毒反应,均不与蝙蝠流感病毒M2的真核表达产物及拯救病毒反应。本研究证实这两种亚型流感病毒的M2蛋白之间确实存在免疫原性的差异,并且筛选到了可以区分蝙蝠流感病毒和禽流感病毒的M2抗体,为进一步研究流感病毒M蛋白相关的生物学机制提供了基础。The homology for M1 protein of the Bat influenza virus and other known Influenza A virus is about 78%, and the homology for M2 protein is much lower as 42%. To reveal the immunogenicity difference for M1 and M2 protein of the newly discovered Bat influenza virus, we constructed the prokaryotic and eukaryotic expression plasmids for the M1/M2 encoded proteins of the H17N10 Bat influenza virus （A/little yellow-shouldered bat/Guatemala/164/2009） and an H9N2 Avian influenza virus（A/chicken/Jiangsu/A2093/2011）. Results showed that the M1 proteins of both viruses expressed by the prokaryotic plasmids were only able to react with the M1 polyclonal antibody, but not two monoclonal antibodies （5F2, 3G8）, whereas the M1 proteins of both viruses expressed by the eukaryotic plasmids or the virus-infected cells only react with the 5F2 monoclonal antibody, not with the 3G8 and polyclonal antibodies. The data indicated the M 1 proteins from both viruses displayed similar immunogenicities of tested antibodies. Interestingly, M2 polyclonal antibody could only react with the H9N2 M2 proteins expressed by eukaryotic plasmids or the virus-infected cells, not with Bat influenza virus. In contrast, the M2 monoclonal antibody（14C2） only reacted with the Bat influenza virus M2 expressed by the prokaryotic plasmids, not with the H9N2 M2 expressed by the prokaryotic plasmids. Our study demonstrated the different immunogenicities for M2 protein between Bat influenza and H9N2 Avian influenza viruses, and revealed that the M2 antibody could be used to distinguish these two viruses. Our study provided a basis for future research related to M proteins of both infuenza viruses.

关 键 词：蝙蝠流感病毒 H9N2亚型禽流感病毒 M蛋白 免疫原性 

分 类 号：S852.659.5[农业科学—基础兽医学]