摘要
目的预测和鉴定梅毒螺旋体(Treponema pallidum,Tp)Tp92蛋白的Th细胞表位,为深入探讨这些表位在梅毒表位疫苗中的作用奠定基础。方法采用RANKPEP和SYFPEITHI软件分析工具联合预测Tp92蛋白的Th细胞表位,人工合成5条表位多肽,以5条表位多肽(同时设Con A阳性对照和RPMI-1640阴性对照)分别刺激梅毒患者和正常人外周血单个核细胞(PBMC),用CCK-8法鉴定预测的Th细胞表位,进一步以ELISA检测INF-γ和IL-4的分泌水平,区分Th细胞表位的类型。结果软件预测显示,Tp92蛋白的P1(103-118AA)、P2(694-712AA)、P3(668-680AA)、P4(300-313AA)、P5(396-410AA)氨基酸序列最可能为其Th细胞表位。CCK-8法检测显示,P2、P3、P5能诱导梅毒患者淋巴细胞明显增殖,而正常人淋巴细胞不反应;ELISA检测细胞因子产生情况发现:P2、P3、P5可刺激梅毒患者淋巴细胞产生较高水平IFN-γ,而不产生IL-4。结论 P2、P3、P5为Tp92蛋白潜在的HLA-DRBl限制性特异性Th1型细胞表位。
To predict and identify the Th-cell epitopes of Treponema pallidum（Tp） Tp92 protein and provide a basis for further discussion on roles of these epitopes in epitope-based syphilis vaccine design, the Th cell epitopes of the protein Tp92 were analyzed and predicted with RANKPEP and SYFPEITHI software, and 5 peptides containing predicted epitopes were artificially synthesized. The five predicted peptides of Tp92 were in vitro co-incubated with peripheral blood mononuclear cells（PBMCs） isolated from syphilis patients and normal humans respectively, with Con A as positive control and RPMI-1640 as negative control. Then the proliferations of PBMCs were detected with CCK-8; the concentrations of INF-γ and IL-4 in the culture media were determined by ELISA kit. Comprehensive meta-analysis of software showed that P1（103-118AA）, P2（694-712AA）, P3（668-680AA）, P4（300-313AA） and P5（396-410AA） of the Tp92 might be the T-cell epitopes. The results of T cell proliferation tests indicated that P2, P3 and P5 could induce PBMCs from syphilis patients to proliferate, but not for the PBMCs from normal humans. And P2, P3 and P5 could induce PBMC from syphilis patients to produce IFN-γ, but not IL-4.Token together, the P2, P3 and P5 might be the HLA-DRBl-restricted Th1 epitopes of Tp92 protein.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2015年第12期1059-1062,共4页
Immunological Journal
基金
国家自然科学基金(81273322)
湖南省教育厅资助科研项目(13C876)
郴州市科技局资助科研项目(2012CJ126)
