摘要
目的:预测并筛选鉴定狂犬病毒糖蛋白及核蛋白BALB/c小鼠MHC限制性CTL和Th表位。方法:从NCBI数据库中获取狂犬病毒糖蛋白及核蛋白的完整氨基酸序列,然后运用生物信息学软件BIMAS、SYFPEITHI和RANKPEP对CTL表位及Th表位进行预测并合成相关抗原表位。BALB/c小鼠经狂犬疫苗免疫后,通过淋巴细胞增殖实验对预测的多肽进行初步筛选。初步筛选的多肽再利用ELISPOT和流式细胞法进行进一步筛选,检测指标主要包括细胞因子IFN-γ和IL-4的分泌水平以及多肽对CD3+CD4+/CD3+CD8+T细胞亚群的影响情况。结果:经软件预测获得8条可能的CTL与Th表位,多肽合成后经分析各条多肽纯度大于90%。实验验证后证明2条多肽G367-381及G333-341能够成为细胞表位,其中G367-381作为Th表位,G333-341作为CTL表位。结论:G367-381及G333-341是狂犬病毒的细胞表位,可用于未来狂犬病毒表位疫苗的研究。
Objective：To predict and identify the murine MHC restricted Th and CTL epitopes of the glycoprotein and nucleoprotein of rabies virus.Methods： The complete amino acid sequences of glycoprotein and nucleoprotein of rabies virus were acquired from NCBI data base.The CTL and Th epitopes were predicted by bioinformatics software BIMAS,SYFPEITHI and RANKPEP.The predicted epitopes were initially identified by the proliferation of lymphocytes in BALB/c mice immunized with rabies virus vaccines.The peptides were further analyzed by ELISPOT and FACS to test their abilities in influencing the secretion of IFN-γ and IL-4 as well as the proliferation of CD3＋CD4＋/ CD3＋CD8＋T cells.Results： Eight epitopes were predicted by the software and the purity of synthesized peptides were greater than 90%.The experiments showed that G367-381 and G333-341 could be the Th epitope and the CTL epitope,respectively.Conclusion： G367-381 and G333-341 were the epitopes of the rabies virus and they can be future used for the design and development of rabies epitope vaccines.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2013年第7期736-740,共5页
Chinese Journal of Immunology
基金
“重大新药创制”科技重大专项(2009ZX09203-701)
“艾滋病和病毒性肝炎等重大传染病防治”国家科技重大专项(2012ZX10002006-002-003)
国家自然基金(30901315)资助
