H1N1流感病毒血凝素异嗜性抗原表位分析及突变研究

项目名称： H1N1流感病毒血凝素异嗜性抗原表位分析及突变研究

项目编号： No.81202373

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学免疫学、法医学

项目作者： 李慧瑾

作者单位： 西安交通大学

项目金额： 23万元

中文摘要： 甲型H1N1流感是一种急性呼吸道传染病，接种疫苗是其主要预防手段。但甲流疫苗接种后所引起的少数严重不良反应增加了人们对其安全性的担心，目前其不良反应的发生机制仍不清楚。我们前期研究发现3株H1N1流感病毒血凝素（HA）单克隆抗体可与人体多种组织抗原发生反应，提示HA和人体组织存在共同抗原表位（异嗜性抗原），推测其可能与疫苗的不良反应有关。本课题拟以这3株单抗为研究对象，首先采用基因工程手段克隆其轻重链可变区基因并测序，利用计算机模拟预测HA与单抗结合的氨基酸位点；然后通过点突变技术改造HA的异嗜性表位，并对HA突变体进行真核表达；最后对突变体的中和、血凝及异嗜性表位进行鉴定，确定去除异嗜性表位对HA的中和表位和血凝表位的影响。本项目的完成，可确定H1N1流感病毒HA异嗜性表位的氨基酸位点，为开发安全、高效、基于HA蛋白突变体的H1N1流感疫苗奠定研究基础，并为其它疫苗的安全性研究提供思路。

中文关键词： 甲型H1N1流感；血凝素；异嗜性表位；疫苗；

英文摘要： Influenza A (H1N1) is caused by the H1N1 influenza virus in acute respiratory infections. A flu vaccine inoculation is the primary means of preventing influenza. A small number of serious adverse reactions caused by H1N1 vaccine making people worry, its mechanism is unclear. Preliminary studies by our group found that three hemagglutinin (HA) monoclonal antibodies of H1N1 influenza virus reaction with human multiple tissue antigens, suggesting that such antibodies may be related to few serious adverse reaction after a flu vaccine vaccination. This topic is intended to be three monoclonal antibodies of H1N1 influenza virus HA with resistance to host activity as research tools, firstly, cloning light and heavy chain variable region of three antibody genes by means of genetic engineering, predicting the amino acid sites of the HA monoclonal antibody binding with HA protein by computer simulations; secondly, knockout the HA heterophil antigen epitope by PCR mutations, eukaryotic expression of different HA mutant protein; Finally, expressing the HA mutated protein by eukaryotic expression system, to determing whether removal of heterophil epitope affect neutralizing epitopes and hemagglutinin epitope. The completion of the project, we can determine the HA epitope heterophilic amino acid sites of H1N1 influenza virus,

英文关键词： influenza A(H1N1)；hemagglutinin；heterophil epitope；vaccine；