抗H7N9流感病毒的单克隆抗体与胃组织的交叉反应研究  

Antibodies against H7N9 influenza virus cross-react with gastric tissue

作　　者：孙晶莹[1,2] 张海祥 冯杨萌[1,2] 封青 李研 孙丽君[1,2] 徐翠香 SUN Jingying;HANG Haixiang;FENG Yangmeng;FENG Qing;LI Yan;SUN Lijun;XU Cuixiang(Shaanri Provincial Key La-boratory of Infection and Immune Diseases,Shaanri Provincial People's Hospital,Xi'an 710068,China;Shanci Engineering Research Center of Cell Immunology,Xi'an 710068,China)

机构地区：[1]感染与免疫疾病重点实验室·陕西省人民医院,陕西西安710068 [2]陕西省细胞免疫工程技术研究中心,陕西西安710068

出　　处：《中国实验诊断学》2024年第9期1107-1111,共5页Chinese Journal of Laboratory Diagnosis

基　　金：陕西省自然科学基础研究计划(2023-JC-QN-0844);陕西省人民医院人才支持计划项目(2023JY-27);陕西省人民医院人才支持计划项目(2021JY-32);陕西省重点研发计划项目(2021ZDLSF01-03)。

摘　　要：目的筛选鉴定与胃组织交叉反应的抗H7N9流感病毒的单克隆抗体,初步探讨流感病毒感染会引发肠道相关疾病的可能致病机制。方法利用H7N9流感病毒制备了大量的单克隆抗体,对制备的单克隆抗体运用ELISA方法进行亚型和效价测定,免疫组化对制备的单克隆抗体与组织的反应性进行筛选,巢式PCR分子生物学技术克隆单克隆抗体轻重链可变区序列。结果制备的单克隆抗体H7N9-71和H7N9-78均能与H7N9抗原特异性结合,抗体效价达1∶10~5,免疫组化结果表明H7N9-71、H7N9-78与人的胃组织能发生特异性反应,进一步的研究发现这两株单抗与小鼠胃组织能特异性结合。轻重链可变区序列结果证实二者为两株不同的单抗。结论H7N9流感病毒可能与胃组织之间存在异嗜性抗原,提示流感病毒感染可能与胃肠道疾病的发生存在某些关联。Objective The monoclonal antibodies against H7N9 influenza virus that cross-react with gastric tissue were screened and identified,and the possible pathogenic mechanism of enteric-related diseases caused by influenza virus infection was preliminarily investigated.Methods We prepared a large number of monoclonal antibodies using H7N9 influenza virus.The subtype and titers of the prepared monoclonal antibodies were determined by ELISA method,the reactivity of the prepared monoclonal antibodies was screened by immunohistochemistry,and the sequence of the variable region of the light and heavy chain of the monoclonal antibodies was cloned by nested PCR molecular biology.Results The prepared monoclonal antibodies H7N9-71 and H7N9-78 can bind specifically to H7N9 virus,with antibody titer of 1∶10~5.Immunohistochemical results showed that H7N9-71 and H7N9-78 could bind specifically to human stomach tissue and mouse stomach tissue.The sequence results of light and heavy chain variable region confirmed that the two strains were different monoclonal antibodies.Conclusion These results indicate that there may be heterophilic antigen between H7N9 influenza virus and gastric tissue,suggesting that influenza virus infection may be associated with the occurrence of gastrointestinal diseases.

关 键 词：H7N9流感病毒 单克隆抗体 交叉反应 抗体可变区 

分 类 号：R373.13[医药卫生—病原生物学]