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目的预测并鉴定肿瘤转移相关抗原肝素酶来源的HLA-A2.1限制性CTL表位。方法采用超基序、量化基序和分子模拟相结合的方法,对肝素酶HLA-A2.1限制性CTL表位进行预测,合成候选表位肽;利用T2细胞的特点,对合成的候选肽与HLA-A2.1分子进行亲和力分析,初步验证预测结果;利用标准51Cr释放试验检测特异性CTLs诱导活性。结果在所筛选的5个候选CTL表位中,Hpa(525-533)、Hpa(277-285)、Hpa(405-413)可在体外有效诱导肝素酶特异性CTLs的产生,对肝素酶阳性且HLA-A2阳性的KATO-Ⅲ细胞具有明显的杀伤效应。结论Hpa(525-533)、Hpa(277-285)、Hpa(405-413)有可能是肿瘤抗原肝素酶HLA-A2.1的限制性CTL表位。
Objective To predict and identify the HLA-A2.1-restricted CTL epitopes of tumor metastasis-associated antigen heparinase. Methods The HLA-A2.1 restricted CTL epitopes of heparanase were predicted by a combination of super-motifs, quantitative motifs and molecular modeling. Candidate epitope peptides were synthesized. Based on the characteristics of T2 cells, Peptide affinity analysis with HLA-A2.1 molecules, preliminary validation of the predictive results; standard 51Cr release test was used to detect specific CTLs induced activity. Results Hpa (525-533), Hpa (277-285) and Hpa (405-413) were effective in inducing heparanase-specific CTLs production in vitro in 5 candidate CTL epitopes, Enzyme positive and HLA-A2 positive KATO-III cells have a clear killing effect. Conclusion Hpa (525-533), Hpa (277-285) and Hpa (405-413) are likely to be the restricted CTL epitopes of tumor antigen heparin HLA-A2.1.