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.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
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)
