基于免疫信息学技术的雏沙门菌多表位疫苗构建

doi:10.16431/j.cnki.1671-7236.2024.01.031

摘要/Abstract

摘要： 【目的】设计针对雏沙门菌（Salmonella Pullorum） IpaJ蛋白的多表位疫苗（multi-epitope vaccine，MEV），为净化鸡白痢提供新的疫苗。【方法】选用IEDB预测雏沙门菌IpaJ蛋白的T淋巴细胞主要组织相容性复合体Ⅰ（MHCⅠ）类分子结合表位；用NetMHCIIpan 4.0 Server预测T淋巴细胞MHC Ⅱ类分子结合表位；用IEDB预测B淋巴细胞表位。将各个服务器预测结果筛选出的表位经过VaxiJen v 2.0评估抗原性后，将合格的表位通过柔性linker串联成多表位疫苗。对构建的多表位疫苗进行抗原性、理化性质、N-糖基化位点、二级结构和三级结构预测。使用分子对接评估多表位疫苗与免疫受体的结合能力，使用免疫模拟评估多表位疫苗的免疫效果，最后优化密码子便于克隆表达。【结果】经筛选后，选择4个MHC Ⅰ、4个MHC Ⅱ和8个B淋巴细胞表位优势表位构建的多表位疫苗MEV-IpaJ。多表位疫苗MEV-IpaJ的分子质量为28.18 ku，为稳定亲水蛋白，具有良好的抗原性，存在4个潜在的N-糖基化位点，α-螺旋、延长链、β-转角和无规则卷曲分别占20.38%、19.23%、8.08%和52.31%。三级结构Ramachandran作图显示，优势区域中含有残基数占89.9%%，进行细化后优势区域的残基数增加到94.0%，三级结构突出表位作图也证明MEV-IpaJ具有良好的免疫原性。分子对接结果显示，MEV-IpaJ与Toll样受体2（Toll-like receptor 2，TLR2）和TLR4蛋白分子可对接。免疫模拟结果显示，MEV-IpaJ具有较好的免疫应答，能提高部分细胞因子的表达，经密码子优化确保设计的MEV-IpaJ可在大肠杆菌K12表达系统中高效、稳定地表达。【结论】本研究构建的多表位疫苗MEV-IpaJ可有效表达并可能诱导强烈的T细胞和B细胞免疫应答。本研究为雏沙门菌多表位疫苗的设计提供了一种新的方法，为雏沙门菌多表位疫苗的研发提供了理论依据及数据支持。

关键词: 雏沙门菌; 免疫信息学; 表位; 多表位疫苗

Abstract: 【Objective】 This study was aimed to design multi-epitope vaccine against IpaJ protein of Salmonella Pullorum and provide a new vaccine for purifying chicken dysentery.【Method】 In this study, IEDB was used to predict the major histocompatibility complex class Ⅰ (MHC Ⅰ) molecular binding epitopes of Salmonella Pullorum IpaJ protein in T lymphocytes.NetMHCIIpan 4.0 Server was used to predict T lymphocytes MHC Ⅱ molecular binding epitopes.B lymphocyte epitope was predicted by IEDB.After the antigenicity of the selected epitopes was evaluated by VaxiJen v 2.0, the qualified epitopes were concatenated into multi-epitope vaccine by flexible linker.Antigenicity, physicochemical properties, N-glycosylation sites, secondary structure and tertiary structure of the constructed multi-epitope vaccine were predicted.Molecular docking was used to evaluate the binding ability of the multi-epitope vaccine to the immune receptor.Immune simulation was used to evaluate the immune effect of the multi-epitope vaccine.Finally, the codon was optimized for cloning expression.【Result】 After screening, the constructed multi-epitope vaccine MEV-IpaJ contained 4 MHC Ⅰ, 4 MHC Ⅱ and 8 B lymphocyte epitope dominance.The relative molecular weight of the multi-epitope vaccine MEV-IpaJ was 28.18 ku, which was a stable hydrophilic protein and had good antigenicity.There were four potential N-glycoylation sites, including α-helix, extended strand, beta turn and random coil accounted for 20.38%, 19.23%, 8.08% and 52.31%, respectively.Ramachandran mapping of tertiary structure showed that the dominant region contained 89.9% of the residual base, and the residual base of the dominant region increased to 94.0% after refinement.The mapping of tertiary structure prominent epitope also proved that the multi-epitope vaccine had good immunogenicity, and molecular docking showed that the MEV-IpaJ could dock with Toll-like receptor 2 (TLR2) and TLR4 protein molecules.The results of immune simulation showed that the MEV-IpaJ had a good immune response and could improve the expression of some cytokines.Codon optimization ensured the efficient and stable expression of the MEV-IpaJ in E.coli K12 expression system.【Conclusion】 The constructed multi-epitope vaccine MEV-IpaJ could be effectively expressed and might induce strong T cell and B cell immune responses.This study provided a new method for the design of multi-epitope vaccine of Salmonella Pullorum, and provided theoretical basis and data support for the research and development of multi-epitope vaccine of Salmonella Pullorum.

Key words: Salmonella Pullorum; immunoinformatics; epitope; multi-epitope vaccine

中图分类号:

S852.4

谭菊, 王永娟, 郭广富, 赵长菁, 夏爱鸿, 李巨银, 吴敏秋, 王宇航, 覃秋苹. 基于免疫信息学技术的雏沙门菌多表位疫苗构建[J]. 中国畜牧兽医, 2024, 51(1): 312-322.

TAN Ju, WANG Yongjuan, GUO Guangfu, ZHAO Changjing, XIA Aihong, LI Juyin, WU Minqiu, WANG Yuhang, QIN Qiuping. Construction of a Multi-epitope Vaccine Against Salmonella Pullorum by Immunoinformatics Approach[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(1): 312-322.

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