柯萨奇病毒A组10型VP1蛋白线性中和表位的筛选和鉴定  

Screening and Identification of Linear Epitopes on the VP1 Protein of Coxsackievirus A10

作　　者：甘燕媚 何韵怡 瞿颖 吴岳 刘启亮[3] 刘洪波 GAN Yanmei;HE Yunyi;QU Ying;WU Yue;LIU Qiliang;LIU Hongbo(Department of Laboratory Medicine,the Second Affiliated Hospital of Guilin Medical University,Guilin 541199,China;College of Medical Laboratory Science,Guilin Medical University,Guilin 541199,China;College of Intelligent Medicine and Biotechnology,Guilin Medical University,Guilin 541199,China)

机构地区：[1]桂林医学院第二附属医院检验科,桂林541199 [2]桂林医学院医学检验学院,桂林541199 [3]桂林医学院智能医学与生物技术学院,桂林541199

出　　处：《激光生物学报》2024年第2期160-166,共7页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(81660280)。

摘　　要：柯萨奇病毒A组10型(CV-A10)是引起手足口病(HFMD)的常见病原体之一。应用重叠肽法筛选出覆盖CV-A10全长VP1区氨基酸序列的39条候选表位肽段,以间接酶联免疫吸附试验(ELISA)及微量中和抑制试验验证。结果发现,候选表位P6(CV-A10 VP1区第39~53位氨基酸)与CV-A10全病毒抗血清具有高反应性,且在3.91μg/mL的稀释质量浓度时仍具有中和抑制作用,为潜在中和表位。用P6肽免疫小鼠制备相应的抗血清,并以微量中和试验验证其保护能力,结果显示,P6抗血清的几何平均中和效价为1:8.97,可确定P6为CV-A10的中和表位。为了解P6序列在CV-A10型内的保守性,将P6的氨基酸序列与CV-A10各基因型代表株进行比对分析,结果显示,P6在CV-A10型内高度保守,表明P6为CV-A10的广谱性中和表位,可应用于CV-A10表位疫苗的研发。本研究旨在筛选鉴定CV-A10 VP1蛋白上的线性中和表位,为CV-A10表位疫苗的研发和HFMD的防控奠定基础。Coxsackievirus A10(CV-A10)is one of the most common pathogens causing hand,foot and mouth disease(HFMD).The 39 candidate epitope peptides covering the amino acid sequence of the VP1 region of CV-A10 were screened by using the overlapping peptide method and verified by indirect enzyme linked immunosorbent assay(ELISA)and microneutralization inhibition assays.It has been found that candidate epitope P6(amino acid 39~53 in VP1 region of CV-A10)showed high reactivity with CV-A10 virus serum,and still showed neutralizing inhibition effect at 3.91μg/mL dilution mass concentration,which was a potential neutralizing epitope.P6 antiserum was prepared by immunizing mice with P6 peptide,and microneutralization test was used to verify its protective effect.The results showed a 1:8.97 geometric mean neutralizing titer for P6 antiserum,which identified P6 as the neutralizing epitope of CV-A10.In order to understand the conservativeness of P6 sequence in CV-A10,the amino acid sequence of the P6 was compared with the representative strains of CV-A10 genotype,and the results showed that P6 was highly conserved in CV-A10.It indicated that P6 is a conserved linear neutralization epitope of CV-A10,which has the potential to resist the infection of various genotypes of CV-A10,and can be used as a target for the development of CV-A10 epitope vaccine.This study aimed to screen and identify linearly neutralizing epitopes on the CV-A10 VP1 protein and to lay a foundation for the development of CV-A10 epitope vaccine and HFMD prevention.

关 键 词：柯萨奇病毒A组10型 VP1蛋白 线性中和表位 疫苗 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]