结核杆菌抗原Rv0173的HLA-A*0201限制性CTL表位的预测及鉴定

目的 鉴定结核杆菌(Mycobacterium tuberculosis,Mtb)抗原Rv0173中的HLA-A*0201限制性CTL表位.方法 应用数据库SYFPETTHI预测结核杆菌Rv0173中可能存在的HLA-A*0201限制性CTL表位,经流式细胞术分析各抗原肽与HLA-A*0201的亲合力,经时间分辨荧光法检测外周血单个核细胞(PBMCs)对各抗原肽产生的增殖反应,经细胞毒性实验研究各抗原肽诱导的特异性T细胞的细胞毒杀伤活性,逐步鉴定Rv0173的HLA-A*0201限制性CTL表位.结果 位于Rv0173氨基酸序列肽3(21-30aa)和抗原肽7(161-170aa)与HLA-A*0201分子具有较高的亲合力,并都能刺激HLA-A*0201阳性个体PBMCs增殖,并诱导产生特异性杀伤活性.结论 肽3(RLSQSADQYL)(21-30aa)和肽7(LLRGGGLVNL)(161-170aa)是抗原Rv0173上HLA-A*0201限制性CTL的优势表位,可作为结核疫苗设计的候选表位.

Prediction and Identification of HLA-A* 0201 restricted CTL epitopes in Mycobacteriun tuberculosis Rv0173

Objective To identify HLA-A*0201 restricted CTL epitopes in Mycobacterium tuberculosis Rv0173.Methods Firstly,database SYFPEITHI was applied to predict HLAA*0201 restricted CTL epitopes in Mycobacterium tuberculosis antigen Rv0173.Secondly,affinity and stability with T2 cell of the predicted peptides were assayed by FACS.Thirdly,cells proliferation and cytotoxicity induced by peptides were investigated by DELFIA cell proliferation assay and DELFIA EUTDA cytotoxicity test respectively.Results Two epitope peptides,21-30aa and 161-170aa,had high affinity with HLA-A*0201 molecules,stimulated HLA-A*0201 PBMCs to proliferate and induced specific cytotoxicity.Conclusion pep3(RLSQSADQYL)(21-30aa) and pep7(LLRGGGLVNL)(161-170aa) are HLA-A*0201 restricted CTL epitopes of Mtb.Antigen Rv0173,which could be the candidates for TB peptide-vaccine.