首页|甲型流感病毒血凝素HA蛋白保守抗原表位的鉴定

甲型流感病毒血凝素HA蛋白保守抗原表位的鉴定

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流感大流行对人类健康危害严重，流感病毒型别多，易变异，现有疫苗不能诱导机体产生不同亚型间的交叉保护力，造成预防困难，因此，精细定位甲型流感病毒保守抗原表位是研究重点。本研究通过使用单克隆抗体和计算机模拟预测两种技术，采用免疫学方法和生物信息学方法，获得了 4株识别血凝素(HA)保守抗原表位的单克隆抗体，以这4株单抗为研究工具，最终明确HA茎部区域多肽STQNAID(409～415 aa)是甲型流感病毒HA保守区抗原表位，为开发甲型流感通用疫苗提供理论和实验基础。
Identification of Conserved Epitopes of the Hemagglutinin HA Protein on Influenza A virus
Influenza A pandemic is seriously harmful to human health.There are many types of influenza virus and it is easy to mutate.The existing vaccines cannot induce cross-protection between different subtypes,which makes prevention difficult.Frequent mutation of influenza A virus hemagglutinin(HA)antigen is the main cause of influenza pandemic.Therefore,the study of B cell epitope on HA is very important in the prevention and control of influenza virus.In this study,monoclonal antibody(mAb)was pre-pared by hybridoma fusion and screening technique using 2009 H1N1 vaccine as immunogen.The reactivity of mAbs with 5 different subtypes of influenza viruses(H1N1 2009,H3N2,avian influenza H5N1 and H7N9)was detected by indirect ELISA.The amino acid sequences of the antigens recognized by the mAbs were found in GenBank database,and the peptides were designed and synthe-sized by DNAMAN software according to the common amino acid sequences of these antigens.Then different peptides were blocked with influenza mAbs to determine the conserved epitopes recognized by these mAbs,and the interrelationship between the mAbs were analyzed.The distribution of conserved epitopes in HA crystal structure was analyzed by Pymol and PDB viewer software.4 mAbs were obtained to recognize the conserved epitope of HA.Using these 4 mAbs as research tools,STQNAID(409～415 aa)in the stem region of HA was finally identified as the epitope of the conserved region of influenza A virus.ELISA blocking assay showed that 4 antibodies could bind to at least two kinds of epitopes on HA,indicating that there are at least two kinds of epitopes in influenza virus HA.Fine mapping of conserved epitopes of influenza A virus is the focus of research,which provides theoretical and experimental basis for developing universal influenza A vaccine.It also provides a new method for the epitope prediction of other pathogenic micro-organisms.

Influenza A virusHAMonoclonal antibodyELISA blocking assayComputer predictionLocalizationConserved epitope

郭春艳、靳占奎、封青、黄晓燕、徐翠香

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陕西省人民医院感染与免疫疾病重点实验室,陕西西安 710068

陕西省细胞免疫工程技术研究中心,陕西西安 710068

陕西省人民医院骨科,陕西西安 710068

甲型流感病毒 血凝素HA 单克隆抗体 阻断ELISA 计算机预测 定位 保守抗原表位

陕西省重点研发计划一般项目陕西省重点研发计划一般项目陕西省教育厅重点研发计划项目

2024SF-YBXM-156S2024-YF-YBSF-069922JS035

2024

10.19526/j.cnki.1005-8915.20240201

药物生物技术
中国药科大学,中国医药科技出版社,中国药学会

药物生物技术

CSTPCD
影响因子：0.463
ISSN：1005-8915
年,卷(期)：2024.31(2)
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