肝素酶HLA-A2限制性CTL表位肽的筛选及其活性鉴定  

Selection and identification of HLA-A2-restricted CTL epitopes within heparanase

作　　者：余彤[1] 王惠菊 杨建民[1] 方勇[3] 周永列[4] 叶再元 

机构地区：[1]浙江省人民医院消化内科,杭州310014 [2]浙江省胃肠病学重点实验室 [3]浙江大学附属邵逸夫医院肿瘤内科 [4]浙江省人民医院检验科,杭州310014

出　　处：《中华微生物学和免疫学杂志》2008年第8期675-679,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30570816）志谢：本项研究得到重庆第三军医大学全军免疫学研究所吴玉章教授的指导帮助,特此感谢.

摘　　要：目的预测并鉴定新的人乙酰肝素酶（Hpa）抗原的HLA-A2限制性CTL表位，为恶性肿瘤多肽疫苗的免疫治疗提供依据。方法采用SYFPEITHI和BIMAS软件预测方法，对肝素酶HLA—A2限制性CTL表位进行预测，合成候选表位肽；利用1、2细胞特点，对合成的候选肽与HLA—A2分子进行亲和力分析；利用乳酸脱氢酶释放试验检测待检肽特异性CTL诱导活性；利用ELISPOT检测T细胞活性。结果在所筛选的6条候选CTL表位肽中，Hpa（310～318）FLNPDVLDI与HLA—A2分子具有高亲和力，在体外可有效诱导肝素酶特异性CTL的产生，对肝素酶阳性且HLA—A2限制性的HCC—LM6肝癌细胞及SW-480结肠癌细胞具有明显的杀伤效应，且能有效诱导IFN-1分泌，增强免疫活性。结论首次发现Hpa（310～318）FLNPDVLDI可能是肿瘤肝素酶抗原的HLA-A2限制性CTL表位。Objective To select and-identify＇human leukocyte antigen （HLA）-A2-restricted T- cell epitope within heparanase （ Hpa）, and to provide academic bases for peptide-based immunotherapy with malignant tumor. Methods The Hpa sequence was scanned for prediction of the immunogenic peptides- based CTL epitopes using the HLA-binding prediction software SYFPEITHI and BIMAS. Then the affinity of candidate epitopes to HLA-A2 was evaluated by T2 binding test. Activation of T cell was assessed by ELIS- POT and cytotoxictiy by lactate dehydrogenase （ LDH ） release assay. Results Of the six predicted nonapeptides, Hpa（310-318, FLNPDVLDI） showed high affinity of binding to HLA-A2 and led to IFN-γsecretion in vitro. Furthermore, Hpa （310-318） FLNPDVLDI could induce PBMC from a HLA-A2 positive healthy donor to lyse specifically HCC-LM6 and SW-480 cells which expressing both Hpa and HLA-A2. Conclusion The nona-peptide Hpa（310-318）FLNPDVLDI may be an HLA-A2-restricted CTL epitope, which is capable of inducing Hpa-specific CTL in vitro.

关 键 词：肝素酶 CTL表位 HLA—A2限制性 多肽 

分 类 号：R392[医药卫生—免疫学] R730.5[医药卫生—基础医学]