IDO修饰的成纤维细胞培养上清对小鼠T淋巴细胞增殖的影响  

Effects on T cell proliferation by culture supernatant of fibroblast cells modified with mIDO gene

作　　者：罗璐[1] 曾耀英[2] 王箴 王青[2] 赵莉[2] 杨冬梓[1] 

机构地区：[1]中山大学附属第二医院妇产科,广东广州510120 [2]暨南大学组织移植与免疫中心,广东广州510632 [3]深圳市南山区蛇口人民医院,广东深圳518067

出　　处：《细胞与分子免疫学杂志》2007年第12期1122-1125,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家教育部博士点基金资助项目(20050558093);中国博士后基金资助项目(20060390755);卫生部部属(管)医疗机构临床学科重点项目(2004-468)

摘　　要：目的:检测吲哚胺2-3双加氧酶(IDO)基因修饰的细胞对T淋巴细胞的免疫抑制作用。方法:将pEGFP-Nl-mIDO真核表达载体转染至小鼠NIH3T3成纤维细胞系。体外检测mIDO修饰的成纤维细胞培养液中色氨酸水平的变化;使用CFSE标记技术,检测mIDO修饰的成纤维细胞培养液对ConA刺激的小鼠T淋巴细胞增殖的影响。结果:将pEGFP-NI-mIDO转入NIH3T3成纤维细胞,流式细胞术(FCM)检测到EGFP的表达;体外实验表明,成纤维细胞培养上清和转染pEGFP-N1的成纤维细胞培养上清可检测到一定量的色氨酸水平,而转染pEGFP-N1-mIDO的成纤维细胞培养上清未检测到色氨酸,提示pEGFP-NI-mIDO转染的成纤维细胞表达了具有活性的mIDO;使用CFSE标记技术,检测mIDO修饰的成纤维细胞培养上清对小鼠T淋巴细胞增殖的影响,发现ConA刺激的IDO组,72h后亲代细胞占各代细胞的百分率明显高于对照组。结论:IDO基因修饰的成纤维细胞可抑制T淋巴细胞的增殖。AIM： To investigate the effects on T cell proliferation by culture supernatant of fibroblast cells modified by mIDO gene. METHODS： Eukaryotic expression plasmid pEGFP-NI-mIDO was transfected into the mouse fibroblast call lines by liposome. The concentration of tryptophan in pEGFP-NI-mIDO fibreblast cell culture medium was tested. The effects on ConA-stimulated T cells, proliferation by pEGFP-NI-mIDO fibroblast cell culture medium were analyzed by CFSE technique. RESULTS. pEGFP-NI-mIDO was transfected into the fibroblasts cell lines successfully. Tryptophan in pEGFP-NI-mIDO-transfected-fibreblast cell culture medium was undetectable, which showed pEGFP- NI-mIDO-transfected-fibreblast cells produced active mIDO, resulting in the degradation of tryptophan in the medium. mlDO could inhibit proliferation of ConA-stimulated T cells, which was marked by CFSE. （CONCLUSION= T cell proliferation can be inhibited by the fibreblast cells modified by mIDO gene.

关 键 词：IDO基因 T细胞增殖 免疫抑制 CFSE 

分 类 号：R392.12[医药卫生—免疫学]