Efficient Identification of Tembusu Virus CTL Epitopes in Inbred HBW/B4 Ducks Using a Novel MHC Class I-Restricted Epitope Screening Scheme

文献类型： 外文期刊

第一作者： Zhang, Lin

作者： Zhang, Lin;Ren, Shuaimeng;Xu, Minli;Wu, Jiaqiang;Zhang, Nianzhi;Li, Zhuolin;Tang, Ziche;Wei, Xiaohui;Xie, Xiaoli;Han, Lingxia;Chen, Hongyan;Ren, Shuaimeng;Meng, Kai;Liu, Yueyue;Qi, Lihong;Wu, Jiaqiang;Wu, Jiaqiang;Zhang, Nianzhi

作者机构：

期刊名称：JOURNAL OF IMMUNOLOGY （ 影响因子：5.426； 五年影响因子：6.178 ）

ISSN： 0022-1767

年卷期： 2022 年 209 卷 1 期

页码：

收录情况： SCI

摘要： The identification of MHC class I-restricted CTL epitopes in certain species, particularly nonmammals, remains a challenge. In this study, we developed a four-step identification scheme and confirmed its efficiency by identifying the Anpf-UAA*76-restricted CTL epitopes of Tembusu virus (TMUV) in inbred haplotype ducks HBW/B4. First, the peptide binding motif of Anpf-UAA*76 was determined by random peptide library in de novo liquid chromatography-tandem mass spectrometry, a novel nonbiased, dataindependent acquisition method that we previously established. Second, a total of 38 TMUV peptides matching the motif were screened from the viral proteome, among which 11 peptides were conserved across the different TMUV strains. Third, the conserved TMUV peptides were refolded in vitro with Anpf-UAA*76 and Anp/-beta(2)-microglobulin to verify the results from the previous two steps. To clarify the structural basis of the obtained motif, we resolved the crystal structure of Anpf-UAA*76 with the TMUV NS3 peptide LRKRQLTVL and found that Asp (34) is critical for the preferential binding of the B pocket to bind the second residue to arginine as an anchor residue. Fourth, the immunogenicity of the conserved TMUV peptides was tested in vivo using specific pathogen-free HBW/B4 ducks immunized with the attenuated TMUV vaccine. All 11 conserved TMUV epitopes could bind stably to Anpf-UAA*76 in vitro and stimulate the secretion of IFN-gamma and lymphocyte proliferation, and three conserved and one nonconserved peptides were selected to evaluate the CTL responses in vivo by flow cytometry and their tetramers. We believe that this new scheme could improve the identification of MHC class I-restricted CTL epitopes, and our data provide a foundation for further study on duck anti-TMUV CTL immunity.

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