流式细胞术不同设门方法对CD4+CD25+调节性T细胞的检测方法比较

目的 比较CD4+CD25hi、CD4+CD25+FoxP3+和CD4+CD25+CD127-/low 3种设门方法对CD4+CD25+调节性T细胞(Treg细胞)的界定效果.方法 完全随机法选择33例本院健康体检者,分别采用CD4+CD25hi、CD4+CD25+FoxP3+和CD4+CD25+CD127-/low设门的方法进行检测.结果 (1)CD4+CD25hi、CD4+CD25+FoxP3+设门方法对CD4+CD25+Treg细胞的检测结果差异无统计学意义[(6.85±2.72)%,(6.69±2.30)%,t=0.270,P=0.788],相关性检验显示,两者呈正相关(r=0.866,P＜0.001);(2)CD4+CD25+CD127-/low设门方法的细胞检测结果[(8.07±2.18)%]高于CD4+CD25hi和CD4+CD25+FoxP3+设门方法(t=2.055,P=0.043;t=2.325,P=0.022),相关性检验显示,均呈正相关(r=0.615,P＜0.001;r=0.683,P＜0.001);(3)CD4+CD25hi细胞群中FoxP3+细胞占(57.11±12.42)%,CD4+CD25hi细胞群中FoxP3+细胞占所有FoxP3+细胞的比例为(54.4±9.0)%.结论 CD4+CD25hi设门方法可用于人外周血CD4+CD25+Treg细胞的检测,CD4+CD25+CD127-/low设门方法可应用于CD4+CD25+Treg细胞的纯化.

Different gating methods for measurement of CD4+CD25+ Treg cells by flow cytometry

Objective To compare the definition effects of CD4+CD25hi, CD4+CD25+FoxP3+ and CD4+CD25+CD127-/low gating methods for measurement of CD4+CD25+ regulatory Treg cells. Methods 33 healthy individuals in our hospital were randomly selected and measured for CD4+CD25+ regulatory Treg cells by flow cytometry with CD4 + CD25hi, CD4 + CD25 + FoxP3 + and CD4 + CD25 + CD127-/low gating methods,method and CD4+CD25+FoxP3+ gating method was not found to be statistically significant [ (6.85±2.72)% ,(6.69±2.30)%, t=0.270, P=0.788]. There was a positive significant correlation between two gating methods 2.18) % ] was higher than those with CD4+CD25hi and CD4+CD25+FoxP3+ gating methods (t=2.055, P=0.043;t=2.325, P=0.022). CD4+CD25+CD127-/low gating method was positively correlated with the other two gating was (57.11 ± 12.42) % accounting for (54.4±9.0)% of all the FoxP3 + cells. Conclusion CD4+CD25hi gating method can be used to measure CD4+CD25+ Treg cells in human peripheral blood, whereas the CD4+CD25+ CD127-/low gating method can be favorable for purification of CD4+CD25+ Treg cells.