MUC4 HLA-A*0201限制性CTL表位的预测及结合力分析

目的：寻找并鉴定MUC4 HLA-A^＊0201限制性细胞毒性T细胞（CTL）表位.方法：结合2种常用表位预测数据库（SYFPEITHI和ProPred-I）和工具,应用多种参数和方法进行综合分析,包括MHC结合力、蛋白酶体切割位点等综合预测方法.预测得到的抗原表位合成相应抗原肽.用T2细胞株测定各肽与HLA-A^＊0201分子的亲和力.结果：综合分析预测得到HLA-A^＊0201的5个可能表位;合成抗原肽经纯化后纯度＞95%;在5个候选表位肽中,LLLGVGTFV（u25-33）和LLGVGTFVV（1126-34）2个九肽与HLA-A^＊0201的结合力较强.结论：多参数表位预测结果和结合力分析实验结果一致性较好,两者联合应用初步认为LLLGVGTFV（1125-33）和LLGVGTFVV（1126-34）2个九肽为MUC4 HLA-A^＊0201限制性CTL表位的可能性最大.

Prediction and peptide binding analysis of HLA-A*0201-restricted CTL epitope from MUC4

Objective: To predict and identify HLA-A*0201-restricted cytotoxic T lymphocyte epitope from MUC4.Methods: 2-epitope prediction databases(SYFPEITHI and ProPred-I) combined with several parameter and methods were used to analyzed.MHC binder and proteasome/immunoproteasome cutting site.T2 cell line was used to determine the peptide binding with HLA-A*0201 molecule.Results: After comprehensive analysis,5 candidate HLA-A*0201 epitopes were acquired.Among five predicted epitopes,LLLGVGTFV(1125-33) and LLGVGTFVV(1126-34) were determined as the two with more HLA-A*0201 affinity.Conclusion: Our results suggest that multi-parameter prediction accorded with peptide binding assay.In this study,both epitope prediction and peptide binding assay show that LLLGVGTFV(1 125-33) and LLGVGTFVV(1 126-34) may be the HLA-A*0201-restricted epitope from MUC4.