中国循证儿科杂志2017，Vol.12Issue(5)：383-387,5.DOI:10.3969/j.issn.1673-5501.2017.05.013
肠道病毒71型VP1抗原表位多肽疫苗的实验研究
Experimental study of the epitope peptide vaccine against the structural protein VP1 of enterovirus 71 strain
摘要
Abstract
Objective To make the epitope peptide vaccine by using the structural protein VP1 of enterovirus 71 (EV71) strain isolated from Guizhou area and investigate the immune protective effects on mice.Methods The VP1 amino acid sequences of 17 EV71 strains isolated from throat swab samples of children patients with hand-foot-mouth disease between March 1,2013 and December 31,2015 in Guizhou area were consistent(a total of 297 amino acids).Peptide fragments were designed and synthesized by using the amino acid sequence,that the last one was 27 amino acids,the others were 20 amino acids.The epitope peptide vaccine was made by using the polypeptide fragments with better immunogenicity,which was screened out by indirect ELISA method and IgG positive serum of children who recovered from EV71 infection.Then the epitope peptide vaccine was used to protect healthy ICR female rats and the control group was set that female rats weren't inoculated (con).The neonatal rats were randomly divided into 2 subgroups of injected and non injected EV71 virus.The mice were killed 2 days after injection of the virus,then the tissue was harvested from the skeletal muscle,the small intestine and brain of all neonatal rats which was used to check EV71 virus by RT-PCR and agarose gel electrophoresis and observed the pathological lesion by H&E.Results 3 peptide vaccines were made by using polypeptide fragments with better immunogenicity screened from 19 segments polypeptide fragments with cross sequences.The female mice were immunized by using the peptide vaccines and the subsequent experiments were carried out in neonatal rats.RTPCR results showed that EV71 virus could be detected in infected groups,could not be detected in uninfected groups.The pathological lesion showed that skeletal muscle,intestine and brain tissue had obvious inflammatory changes in the neonatal rats infected with EV71 virus but the female mice weren't immunized.Inflammatory changes were significantly alleviated in the neonatal rats infected with EV71 virus and the female mice were immunized,there was no significant difference in each tissue between the three vaccinated groups.No obvious inflammatory lesion was found in each tissue of the neonatal rats which didn't infected with EV71 virus and the female mice were immunized.Conclusion In our study,VP1 epitope peptide vaccine was successfully prepared,and its efficacy and safety were confirmed in animal experiments,it can provide methodological reference for the development of related vaccines in the future.关键词
手足口病/肠道病毒EV71/结构蛋白VP1/多肽疫苗/贵州Key words
hand-foot-mouth disease/enterovirusEV71/VP1/Epitope peptide vaccine/Guizhou引用本文复制引用
向云凤,宋惠玲,苟恩进,李青,顾胜利,韩允,汤正珍,黄波..肠道病毒71型VP1抗原表位多肽疫苗的实验研究[J].中国循证儿科杂志,2017,12(5):383-387,5.基金项目
国家自然科学81260292 （）
遵义市科技局科技计划基金项目:遵市科合社字[2013]14号 （）
遵义市科学技术基金项目遵市科合人才[2015]24号 （）
遵义市“15851人才精英工程”培养人才在研项目经费资助项目:2013-20 （）
