梅毒螺旋体Tp92蛋白T细胞表位的预测与鉴定
发布时间：2018-10-20 17:43
【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。
[Abstract]:Objective to predict and identify the Th cell epitopes of Tp92 protein of Treponema pallidum (Treponema pallidum,Tp), and to lay a foundation for further study of the role of these epitopes in syphilis epitope vaccine. Methods RANKPEP and SYFPEITHI software were used to predict the Th cell epitopes of Tp92 protein, and 5 epitope peptides were synthesized. Five epitope polypeptides (Con A positive control and RPMI-1640 negative control) were used to stimulate (PBMC), of peripheral blood mononuclear cells from syphilis patients and normal subjects respectively. The predicted Th epitopes were identified by CCK-8 method. The secretion levels of INF- 纬 and IL-4 were further detected by ELISA. Identify the types of Th cell epitopes. Results the amino acid sequences of P1 (103-118AA), P2 (694-712AA), P3 (668-680AA), P4 (300-313AA), P5 (396-410AA) of Tp92 protein were most likely to be their Th epitopes. The results of ELISA analysis showed that P2P3P5 could stimulate the production of IFN- 纬 in the lymphocytes of syphilis patients, but not the production of IL-4. in the patients with syphilis. Conclusion P _ (2) P _ (3) P _ (5) is a potential HLA-DRBl restricted specific Th1 epitope of Tp92 protein.
【作者单位】： 南华大学医学院病原生物学研究所;湘南学院预防医学与医学检验系;
【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)
【分类号】：R759.1
[Abstract]:Objective to predict and identify the Th cell epitopes of Tp92 protein of Treponema pallidum (Treponema pallidum,Tp), and to lay a foundation for further study of the role of these epitopes in syphilis epitope vaccine. Methods RANKPEP and SYFPEITHI software were used to predict the Th cell epitopes of Tp92 protein, and 5 epitope peptides were synthesized. Five epitope polypeptides (Con A positive control and RPMI-1640 negative control) were used to stimulate (PBMC), of peripheral blood mononuclear cells from syphilis patients and normal subjects respectively. The predicted Th epitopes were identified by CCK-8 method. The secretion levels of INF- 纬 and IL-4 were further detected by ELISA. Identify the types of Th cell epitopes. Results the amino acid sequences of P1 (103-118AA), P2 (694-712AA), P3 (668-680AA), P4 (300-313AA), P5 (396-410AA) of Tp92 protein were most likely to be their Th epitopes. The results of ELISA analysis showed that P2P3P5 could stimulate the production of IFN- 纬 in the lymphocytes of syphilis patients, but not the production of IL-4. in the patients with syphilis. Conclusion P _ (2) P _ (3) P _ (5) is a potential HLA-DRBl restricted specific Th1 epitope of Tp92 protein.
【作者单位】： 南华大学医学院病原生物学研究所;湘南学院预防医学与医学检验系;
【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)
【分类号】：R759.1
【参考文献】
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