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Identification of Unique and Conserved Neutralizing Epitopes of Vestigial Esterase Domain in HA Protein of the H9N2 Subtype of Avian Influenza Virus

文献类型： 外文期刊

作者： Huang, Xiangyu 1 ; Yin, Guihu 1 ; Cai, Yiqin 1 ; Hu, Jianing 1 ; Huang, Jingwen 1 ; Liu, Qingtao 2 ; Feng, Xiuli 1 ;

作者机构： 1.Nanjing Agr Univ, Coll Vet Med, Chinas Minist Agr, Key Lab Anim Microbiol, Nanjing 210095, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Vet Med, Minist Agr, Key Lab Vet Biol Engn & Technol, Nanjing 210014, Peoples R China

3.Nanjing Agr Univ, Coll Vet Med, MOE Joint Int Res Lab Anim Hlth & Food Safety, Nanjing 210095, Peoples R China

关键词： HA protein; neutralization; monoclonal antibody; antigen epitope

期刊名称：VIRUSES-BASEL （ 影响因子：5.818； 五年影响因子：5.811 ）

ISSN：

年卷期： 2022 年 14 卷 12 期

页码：

收录情况： SCI

摘要： The H9N2 subtype of avian influenza virus (AIV) has been reported to infect not only birds, but also humans. The hemagglutinin (HA) protein is the main surface antigen of AIV and plays an important role in the viral infection. For treatment strategies and vaccine development, HA protein has been an important target for the development of broadly neutralizing antibodies against influenza A virus. To investigate the vital target determinant cluster in HA protein in this work, HA gene was cloned and expressed in the prokaryotic expression vector pET28a. The spleen lymphocytes from BALC/c mice immunized with the purified recombinant HA protein were fused with SP2/0 cells. After Hypoxanthine-Aminopterin-Thymidine (HAT) medium screening and indirect ELISA detection, six hybridoma cell lines producing anti-HA monoclonal antibodies were screened. The gradually truncated HA gene expression and western blotting were used to identify their major locations in epitopes specific to these monoclonal antibodies. It was found that the epitopes were located in three areas: (NVENLEEL119)-N-112, (EELRSLFS124)-E-117, and (170)PIQDAQ(175). Epitope (NVENLEEL119)-N-112 has a partial amino acid crossover with (EELRSLFS124)-E-117, which is located in the vestigial esterase domain "110-helix" of HA, and the monoclonal antibody recognizing these epitopes showed the neutralizing activity, suggesting that the region (NVENLEELRSLFS124)-N-112 might be a novel neutralizing epitope. The results of the homology analysis showed that these three epitopes were generally conserved in H9N2 subtype AIV, and will provide valuable insights into H9N2 vaccine design and improvement, as well as antibody-based therapies for treatment of H9N2 AIV infection.

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