双向电泳-质谱法分析人CD4^+CD25^+T细胞与CD4^+CD25^-T细胞蛋白质组差异

Differential proteomic analysis of human CD4^+CD25^+ vs CD4^+CD25^-T cells by 2-DE

目的 用双向凝胶电泳-质谱法(2-DE/MS)分析人CD4+ CD25+T细胞和CD4+CD25-T细胞表达的蛋白质组,建立二者之间的差异表达谱。方法提取人脾脏源CD4+ CD25+T细胞和CD4+CD25-T细胞总蛋白质,双向电泳分离蛋白,比较两种细胞的蛋白质组表达差异,对差异蛋白质点用基质辅助激光解析电离飞行时间质谱(Matrix-assisted laser des-orption/ionization time of flying mass spectrometry,MALDI-TOF-MS)进行鉴定。结果胶图差异分析发现25个表达水平存在明显差异的蛋白质点,其中17个点在CD4+CD25-T细胞上调,8个点在CD4+CD25+T细胞上调。质谱初步鉴定出15个差异蛋白质点。结论CD4+ CD25+T细胞和CD4+ CD25-T细胞表达蛋白质组存在差异,这种蛋白质组的差异分析有助于进一步研究调节性T细胞功能相关蛋白质。

Objective To analyze comparatively the proteomes expressed by human CD4+ CD25+ T cells and CD4+ CD25- T cells. Methods The difference of protein expression in human splenic CD4+ CD25+ T cells and CD4+CD25- T cells was analyzed by two-dimensional electrophoresis (2-DE) . The differential protein spots were identified by matrix-assisted laser desorption/ionization time of flying mass spectrometry(MALDI-TOF-MS) . Results Gel-image analysis revealed that there were 25 differential protein spots, of which 17 protein spots were detected in CD4+ CD25- T cells while 8 protein spots in CD4+ CD25+ T cells. A total of 15 differential protein spots were identified by MALDI-TOF-MS. Conclusion There is significant difference between proteome of human CD4+ CD25+ T cells and that of CD4+ CD25- T cells. The comparative proteomic data can be used for the studies of function-associated proteins of regulatory T cells.