负载野生型p53抗原肽HLA-A^＊2402四聚体的制备和鉴定  

Preparation and identification of soluble HLA-A^＊2402 tetramer loaded with a wild-type p53 antigenic peptide

作　　者：王笑迎[1] 查庆兵[2] 何贤辉[1] 欧阳东云[1] 徐丽慧[3] 郭贺[1] 高琦[1] 

机构地区：[1]暨南大学生命科学技术学院组织移植与免疫实验中心,广东广州510632 [2]暨南大学附属第一医院产前诊断中心,广东广州510632 [3]暨南大学生命科学技术学院生物工程研究所,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2010年第2期136-139,共4页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(30572199);广东省自然科学基金(8451063201000340)

摘　　要：目的:制备负载野生型p53抗原肽的HLA-A*2402四聚体,用于检测卵巢肿瘤患者p53特异性细胞毒T淋巴细胞(CTL)。方法:在p53125-134抗原肽TYS和轻链β2微球蛋白存在时,利用稀释法复性获得负载p53125-134抗原肽(TYSPALNKMF,TYS)的可溶性单体,经生物素酰化并纯化后与荧光素标记的链亲和素按4∶1的比例混合形成HLA-A*2402/TYS四聚体,以流式细胞仪检测特异性CTL的频率。结果:流式细胞术分析显示,卵巢癌患者外周血中可检测减低频率的p53125-134TYS抗原肽特异性CTL。结论:成功制备HLA-A*2402/TYS四聚体。Aim：In order to detect the p53-specific cytotoxic T lymphocytes（CTL） in ovarian cancer patients,HLA-A^＊2402 tetramer which loaded with wild-type p53 antigenic peptide was prepared. Methods： The soluble HLA-A^＊2402/TYS monomer was refolded in the presence of β2-microglobulin and p53 antigenic peptide（TYSPALNKMF,TYS） with dilution method.After biotinylation and purification,the tetramers were formed by incubating the monomers with streptavidin-PE in a ratio of 4 to 1.Flow cytometry analysis was performed to determine the frequency of binding with specific CTL.Results： Flow cytometry analysis indicated that the reducible frequency of p53125-134 TYS peptide-specific CTL could be detected in the peripheral blood of ovarian cancer patients.Conclusion： HLA-A^＊2402/TYS tetramer was prepared successfully.

关 键 词：HLA-A*2402 四聚体 p53抗原 细胞毒T淋巴细胞 

分 类 号：R392.3[医药卫生—免疫学]