肠道病毒C组96型VP1蛋白线性B细胞表位筛选与鉴定  

Screening and identification of linear B cell epitopes on the VP1 protein of enterovirus C96

作　　者：何韵怡 刘阳阳 胡嘉华 刘洪波 HE Yunyi;LIUYangyang;HU Jiahua;LIU Hongbo(College of Medical Laboratory Science,Guilin Medical University,Guilin 541199,China;Department of Laboratory Medicine,the Second Affiliated Hospital of Guilin Medical University,Guilin 541199,China;Department of Science and Research,the Second Affiliated Hospital of Guilin Medical University,Guilin 541199,China;Guangdong Provincial Center for Disease Control and Prevention,Guangzhou 511430,China)

机构地区：[1]桂林医学院医学检验学院,桂林541199 [2]桂林医学院第二附属医院检验科,桂林541199 [3]桂林医学院第二附属医院科研科,桂林541199 [4]广东省疾病预防控制中心,广州511430

出　　处：《华夏医学》2024年第2期1-9,共9页Acta Medicinae Sinica

基　　金：国家自然科学基金项目(81660280);广西高校中青年教师科研基础能力提升项目(2023KY0516);广西高校大学生创新创业计划项目(202210601052)。

摘　　要：目的鉴定肠道病毒C组96型(enterovirus C96,EV-C96)VP1蛋白的线性B细胞表位。方法采用IEDB网站提供的在线分析工具分析、预测EV-C96 VP1蛋白的线性B细胞表位,结合其二级结构、β-转角、抗原性、亲水性等多项参数,筛选出优势候选表位;应用间接ELISA法检测候选表位与EV-C96免疫血清的反应性以及与21种其他常见手足口病病原血清的交叉反应性;应用微量中和试验测定表位抗体的中和效价;采用序列比对分析表位的序列保守性;应用结构对齐分析表位的结构特异性。结果通过生物信息学筛选出EVC96 VP1蛋白7个候选表位(P1~P7);ELISA法检测发现P7(氨基酸序列位置为282~304)与EV-C96多克隆抗体有强反应性,与其他常见EV多克隆抗体不发生明显反应;微量中和试验显示,P7抗体的中和效价低于1∶2;序列及结构分析结果显示,P7的氨基酸序列在EV-C96型内具有较高保守性,与其他EV相应位点氨基酸序列的结构有明显差异。结论P7表位为EV-C96特异的非中和性B细胞表位,可作为开发EV-C96检测试剂盒的候选抗原表位。Objective To identify linear B cell epitopes of VP1 protein of enterovirus C96(EV-C96).Methods The linear B cell epitopes and the property parameters of EV-C96 VP1 protein were predicted by the IEDB online analysis tools,combined with its secondary structure parameters such asβ-turning angle,antigenicity,and hydrophilicity were used to screen out candidate epitopes with advantages.The reactivity of the candidate epitopes with the EV-C96 antiserum and the cross-reactivity with antiserums against 21 other common enterovirus caused hand foot and mouth disease were determined by indirect ELISA assay.The neutralization titer of epitope antibody was determined by microneutralization test.The conservativeness of epitopes was analyzed by sequence alignment,and the structural specificity of epitopes was analyzed by structural alignment.Results Seven epitopes of EV-C96 VP1 protein(P1~P7)were predicted by bioinformatics methods,among which P7(aa282~304)showed strong reactivity with EV-C96 antiserum and did not cross-react with other common enterovirus antisera.The neutralizing titer of P7 antibody was less than 1∶2.The amino acid sequence of P7 was highly conservative in EV-C96 isolates,and the structure of the P7 was different from other 21 common EVs.Conclusion P7 is a non-neutral EV-C96-specific B cell epitope that can be used as a candidate antigen epitope for the development of EV-C96 detection kits.

关 键 词：肠道病毒C组96型 VP1蛋白 线性B细胞表位 手足口病 

分 类 号：R373.2[医药卫生—病原生物学] R512.5[医药卫生—基础医学]