五株EV71病毒毒力、免疫原性与基因组比较分析  

Comparative analysis of the virulence,immunogenicity and genomic sequences of five EV71 isolates

作　　者：刘鑫[1] 段长松[1] 李仲[1] 王校伟[1] 赵忠鹏[1] 王希良[1] 

机构地区：[1]中国人民解放军军事医学科学院微生物流行病研究所免疫学研究室病原微生物生物安全国家重点实验室,北京100071

出　　处：《免疫学杂志》2011年第6期494-498,共5页Immunological Journal

基　　金：国家自然科学基金(30901204)

摘　　要：目的对5株EV71分离株(LCH01、LCH02、BJ01、BJ02、AH01)进行毒力、免疫原性和基因组的比较分析。方法分别用5株EV71分离株攻击Balb/c乳鼠,比较毒力差异;免疫6～8周龄Balb/c小鼠后收集血清,ELISA检测抗体效价,微量中和实验检测中和抗体效价;反转录获得5株EV71的全长cDNA进行全基因组测序;最后,用CLC Main Workbench 5.5软件对序列进行比较分析。结果通过对VP1基因的比较分析发现,这5株EV71分离株均为C4a亚型。1周龄Balb/c小鼠经脑部注射LCH01株,表现出后肢瘫痪和后继死亡;而注射其它4株EV71的小鼠在观察了14 d之后仍然存活。5株EV71血清抗体效价为210～211,中和抗体效价为27～28,无显著差异。LCH01毒株与其它几株序列相比,在5′-NTR、3′-NTR、3D等处均有明显差异。结论这5株EV71分离株为近几年流行的C4a亚型,具有较高的免疫原性;LCH01毒株的毒力高于其它几株,从基因组水平上找到了差异序列,这为EV71疫苗评价和致病机制研究提供了实验数据和实验材料。We aimed to analysis comparatively the virulence,immunogenicity,and genome of the five EV71 isolates： LCH01,LCH02,BJ01,BJ02,AH01.Firstly,one week old Balb/c mice were challenged by the five EV71 isolates respectively to observe their virulence.Secondly,the Balb/c mice were immunized by the five EV71 antigens and then detected the total IgG titers and the neutralizing antibody titers was analyzed by ELISA and neutralization test.Thirdly,five EV71 full-length cDNA were obtained by reverse transcription and were sequenced.Finally,CLC Main Workbench 5.5 software was applied to do the comparative analysis of the sequences.According to above results,a phylogenetic dendrogram was constructed based on the alignment of the entire VP1 gene sequences,which revealed that all five EV71 isolates grouped into the C4a subtypes.We observed that five EV71 isolates presented different clinical symptoms.One-week-old Balb/c mice inoculated with LCH01 isolate by the intracerebral routes developed hind-leg paralysis and subsequently died,while mice inoculated with other isolates survived throughout the 14-day observation.There is no significant difference between titers of total IgG and neutralizing antibody（210-211 and 27-28 respectively）.The sequences of the 5＇-NTR,3＇-NTR and 3Dpol finger domain were aligned to present differences.As conclusion,these five EV71 strains grouped into the C4a subtypes with high immunogenicity;the genomic differences appeared to explain their different virulence,which might provide a reasonable basis to investigate molecular pathogenesis of EV71.

关 键 词：手足口病 EV71病毒 毒力 免疫原性 序列分析 

分 类 号：R373[医药卫生—病原生物学]