【摘要】 目的寻找TE1细胞MHCⅠ类分子提呈的抗原肽。方法用弱酸洗涤法获得TE1细胞表面与MHCⅠ类分子结合的抗原肽,经SepPakC18柱、Centrion3超滤器和RPHPLC去盐纯化后,用负载该抗原肽的树突状细胞刺激生成肿瘤特异性杀伤细胞,以Cr51释放法测定CTL的杀伤活性。结果高效液相色谱图显示多个峰;负载抗原肽DC刺激的CTL比未负载抗原肽DC刺激的CTL对TE1细胞杀伤率高,两者差异有统计学意义;负载抗原肽DC刺激的CTL对与抗原肽孵育的T2细胞比对未经与抗原肽孵育的T2细胞杀伤率高,两者差异有统计学意义。结论弱酸洗涤法可从TE1细胞上获得有效的抗原肽;混合抗原肽中有可与HLAA2分子结合并能激发CTL的抗原肽。更多还原

【Abstract】 Objective To seek antigen peptides presented by MHC-Ⅰ molecules in human esophageal carcinoma TE-1 cell line. Methods Antigen peptides were obtained from TE-1 cell membranes by citrate-phosphate buffer elution. These peptides were subsequently purified by methods of SepPak-C18, Centrion-3, and RP-HPLC. The tumor-specific cytolytic T lymphocytes (CTL) were generated from T cells activated by dendritic cells loaded with acid-eluted peptides, and the cytotoxicity of CTL was assayed by Cr 51 release method in vitro. Re- sults There were some peaks in the chromatogram. The killing rate of CTL activated by dendritic cells loaded with esophageal cancer antigen to TE-1 was significant higher than that of CTL activated by dendritic cells without esophageal cancer antigen loading. The killing rate of CTL activated by DC loaded with esophageal cancer antigen to T2 cells which were reacted with esophageal cancer was significantly higher than that of CTL activated by DC loaded with esophageal cancer antigen to T2 which were not reacted with esophageal cancer antigen. Conclusion Effective esophageal cancer antigen, which can link with HLA-A2 molecular and generate specific CTL by activating lymphocytes among esopha- geal cancer antigen peptide, is obtained from TE-1 cell membranes by using acid elution.更多还原