角蛋白17上HLA-DRB1*07限制性T细胞表位的确定

Identification of HLA-DRB1*07-restricted T cell epitope on keratin 17

目的对银屑病自身抗原棗角蛋白17的HLA-DRB1*07限制性T细胞表位进行确定.方法在前期已确定角蛋白17的HLA-DRB1*07限制性T细胞表位区试验基础上,借助互联网服务器对其中的T细胞表位进行分析预测,人工合成T细胞表位DNA并进行GST融合表达和纯化;分离纯化外周血T细胞,将各融合表位肽段分别与正常人和银屑病患者的T细胞体外作用,检测T细胞增殖程度和培养上清中1FN-γ水平变化,从而筛选出强反应性表位.结果角蛋白17的S1(118-132)、S2(169-183)、S4(323-337)和S4(348-362)表位能够显著地促进银屑病患者T细胞增殖和IFN-γ分沁,与正常对照组比较差异有统计学意义(P均＜0.001).结论 S1(118～132)(VRALEEANTELEVKI)、S2(169～183)(ANILLQIDNARLAAD)、S4(323～337)(MQALEIELQSOQLSMK)和S4(348～362)(ENRYCVQLSQIQGLI)是银屑病特异性强反应性HLA-DRB1*07限制性T细胞表位.本研究为以后研究银屑病的免疫发病机制和治疗打下了一定基础.

Objective To identify the HIA-DRB1 ＊ 07-restricted T cell epitopes on keratin 17 （K17）, one of the autoantigens of psoriasis. Methods In the previous study we identified the HLA-DRB1 ＊ 07-restricted T cell epitope regions on K17. Epitopes in the regions were predicted with interact servers in this study. The two nucleic acid strains of each predicted epitope with restriction endonuclease sites were synthesized and then expressed at N-temainus of GST. These recombinant epitopes and the level of T cell proliferation and the concentration of IFN-γ in the culture were detected. Results Compared with the control group and other epitopes, S1（118- 132）, S2（169-183）, S4（323-337） and S4（348-362） had a positive role on the proliferation and IFN-γ expression of T cells from psoriatic patients （ P 〈 0. 001 ）. Conclusion The results indicated that epitopes S1 （ 118- 132）（VRALEEANTELEVKI）, S2（169-183） （ANILLQIDNARLAAD） , S4（323-337）（MQALEIELQSQLSMK） and S4（348-362）（EMRYCVQLSQIQCLI） the psoriasis-specific HLA-DRB1 ＊ 07-restricted T cell epitopes. The advanced study targeting to these peptides can provide a more complete understanding of the immunological basis of the disease.