肠道病毒71型灭活疫苗诱导小鼠细胞免疫抗原表位的筛选Screening of antigenic epitopes of inactivated enterovirus 71 vaccine inducing cellular immunity in mice
高帆;王一平;方鑫;毛群颖;朱凤才;杨静玉;程刚;鲁凤民;梁争论;李凤祥;王军志;
摘要(Abstract):
目的筛选肠道病毒71型(Enterovirus 71,EV71)灭活疫苗免疫小鼠后诱导的细胞免疫抗原表位,探讨EV71疫苗诱导的细胞免疫应答机制。方法以EV71 BJ08株(C4基因型)病毒结构蛋白氨基酸序列为模板,合成覆盖EV71 VP1~VP3的256条合成肽(每条肽12个氨基酸),分别以其为刺激物(80μg/ml),采用ELISPOT法测定EV71灭活疫苗免疫小鼠脾单个核细胞(Mononuclear cell,MNC)诱导IFNγ分泌的斑点形成细胞数(Spot-forming cell,SFC),筛选细胞免疫抗原表位。从脾MNC中去除CD4+或CD8+T细胞及封闭MHCⅡ类分子或MHCⅠ类分子,分析抗原提呈途径。结果通过对256条合成肽的筛选,获得3条可刺激小鼠脾MNC高水平分泌IFNγ的合成肽。以3条合成肽作为刺激物时,小鼠脾MNC分泌IFNγSFC均值(单针免疫组:22.3、12.8、17.4;两针免疫组:29.3、44.1、28.5)较乙肝表面抗原对照肽S28-3(9单针:2.1;两针:5.2)显著升高(P<0.01)。当MNC中去除CD4+T细胞或封闭MHCⅡ类分子后,IFNγSFC均值显著下降(P<0.05);而去除CD8+T细胞或封闭MHCⅠ类分子时,IFNγSFC均值未见显著性差异(P>0.05)。结论已成功筛选出3个EV71细胞免疫抗原表位,分别位于VP1、VP2和VP3区域,3条合成肽均属MHCⅡ类限制性多肽。
关键词(KeyWords): 肠道病毒A型,人;疫苗,灭活;免疫,细胞;抗原表位
基金项目(Foundation): 国家科技支撑项目(2008BAI69B01)
作者(Author): 高帆;王一平;方鑫;毛群颖;朱凤才;杨静玉;程刚;鲁凤民;梁争论;李凤祥;王军志;
Email:
DOI: 10.13200/j.cjb.2012.01.6.gaof.013
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