戊型肝炎病毒颗粒性蛋白疫苗H-2^d限制性Th表位的筛选  

Identification of H-2^d Restricted Th Epitope in Hepatitis E Virus Particulate Vaccine HEV 239

作　　者：伍小路[1] 吴婷[1] 欧山海[1] 林春鑫[1] 程通[1] 李少伟[1] 张军[1] 夏宁邵[1] 

机构地区：[1]厦门大学国家传染病诊断试剂与疫苗工程技术研究中心福建省医学分子病毒学研究中心,福建厦门361005

出　　处：《病毒学报》2006年第3期176-179,共4页Chinese Journal of Virology

基　　金：863计划(2005AA2Z3H20);福建省科技重大专项(2004YZ01);厦门市重大产业科技项目(3502Z20041008)

摘　　要：戊型肝炎病毒衣壳蛋白重组抗原HEV239能形成类病毒颗粒，具备演变成多价疫苗的载体的潜力，此文旨在筛选、鉴定其内包含的H-2^4限制性Th表位。以50μg HEV 239蛋白与完全弗式佐剂混合后皮下免疫BALB／c鼠，以覆盖HEV 239蛋白全长的15氨基酸肽库体外刺激其脾细胞，用IFN-γ-ELISPOT方法检测其细胞免疫应答，并通过磁珠剔除脾细胞中CD4^＋ T细胞或CD8^＋ T细胞以分析筛选得到的T细胞表位的特性。结果显示：HEV239中包含优势的T细胞表位P34（HEV PORF2 AA533～AA547，HSKTF FVLPL RGKLS）及数个较弱的T细胞表位。P34对HEV 239免疫的BALB／c鼠脾细胞的刺激效果与HEV 239蛋白相当，剔除实验表明该表位为CD4^＋T细胞表位，即Th表位。Recombinant particulate hepatitis E virus （HEV） vaccine HEV 239 is a potential candidate of poly-valent vaccine vector. In this paper, a dominant H-2^d restricted Th epitope in HEV 239 was identified. Groups of BALB/c mice were subcutaneously （s.c.） immunized twice by 50μg HEV 239 protein mixed with complete Freund＇s adjuvant（239-CFA）. A peptide library covering the whole HEV 239 protein, containing 46 peptides with 10 amino acids overlapping, was synthesized and used to stimulate the spleen ceils separated from the immunized mice in vitro. IFN-γ-ELISPOT assay results showed that P34 （HEV PORF2 533 - 547aa, HSKTF FVLPL RGKLS）could stimulate the spleen cells to produce 413 spots/million spleen cells which was similar to HEV 239 protein. Thus, the peptide P34 was identified as a dominant H-2^d restricted T cell epitope. After depletion of the CD4^＋ T ceils in the immunized spleen ceils by magnetic bead separation, the IFN-γ spots/million spleen cells decreased to the background level while almost no influence was observed after CD8^＋ T cell depletion. This result showed that P34 was a H-2^d restricted Th epitope.

关 键 词：戊型肝炎病毒 T辅助细胞 T细胞表位 

分 类 号：R392[医药卫生—免疫学]