【摘要】：目的預(yù)測和鑒定梅毒螺旋體(Treponema pallidum,Tp)Tp92蛋白的Th細(xì)胞表位,為深入探討這些表位在梅毒表位疫苗中的作用奠定基礎(chǔ)。方法采用RANKPEP和SYFPEITHI軟件分析工具聯(lián)合預(yù)測Tp92蛋白的Th細(xì)胞表位,人工合成5條表位多肽,以5條表位多肽(同時設(shè)Con A陽性對照和RPMI-1640陰性對照)分別刺激梅毒患者和正常人外周血單個核細(xì)胞(PBMC),用CCK-8法鑒定預(yù)測的Th細(xì)胞表位,進(jìn)一步以ELISA檢測INF-γ和IL-4的分泌水平,區(qū)分Th細(xì)胞表位的類型。結(jié)果軟件預(yù)測顯示,Tp92蛋白的P1(103-118AA)、P2(694-712AA)、P3(668-680AA)、P4(300-313AA)、P5(396-410AA)氨基酸序列最可能為其Th細(xì)胞表位。CCK-8法檢測顯示,P2、P3、P5能誘導(dǎo)梅毒患者淋巴細(xì)胞明顯增殖,而正常人淋巴細(xì)胞不反應(yīng);ELISA檢測細(xì)胞因子產(chǎn)生情況發(fā)現(xiàn):P2、P3、P5可刺激梅毒患者淋巴細(xì)胞產(chǎn)生較高水平IFN-γ,而不產(chǎn)生IL-4。結(jié)論 P2、P3、P5為Tp92蛋白潛在的HLA-DRBl限制性特異性Th1型細(xì)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- 緯 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- 緯 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.
【作者單位】： 南華大學(xué)醫(yī)學(xué)院病原生物學(xué)研究所;湘南學(xué)院預(yù)防醫(yī)學(xué)與醫(yī)學(xué)檢驗(yàn)系;
【基金】：國家自然科學(xué)基金(81273322) 湖南省教育廳資助科研項目(13C876) 郴州市科技局資助科研項目(2012CJ126)
【分類號】：R759.1

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相關(guān)期刊論文 前4條

1 宋帥;李春玲;賈愛卿;楊冬霞;李淼;;抗原表位研究方法進(jìn)展[J];動物醫(yī)學(xué)進(jìn)展;2010年12期

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