三色法流式细胞术检测胞内细胞因子的建立  

Determination of Intracellular Cytokines by 3 Color Flow Cytometry

作　　者：胡大伟[1] 陈顺乐[1] 沈南[1] 薛峰[1] 裴军[1] 顾越英[1] 鲍春德[1] 石学耕[2] 史桂英[2] 

机构地区：[1]上海第二医科大学附属仁济医院风湿病科,上海200001 [2]上海第二医科大学生物物理教研室,上海200025

出　　处：《上海免疫学杂志》1999年第5期280-282,共3页Shanghai Journal of Immunology

摘　　要：运用荧光标记的抗细胞表面抗原单抗及抗细胞因子单抗， 结合固定、破膜处理技术， 建立了三荧光染色法流式细胞术检测细胞内细胞因子的方法， 探讨了不同刺激剂及细胞培养时间的选择， 并对５ 例正常人产生ＩＦＮγ、ＩＬ４ 的ＣＤ４ 及ＣＤ８ 细胞进行测定。结果表明： 产生ＩＦＮγ的ＣＤ４ 及ＣＤ８ 细胞明显多于产生ＩＬ４ 的细胞， 并且产生ＩＦＮγ的ＣＤ８ 细胞百分数比较ＣＤ４ 细胞相对增多； 同时产生ＩＦＮγ／ＩＬ４ 的ＣＤ４ 、ＣＤ８ 细胞较少。此方法可从单个细胞水平直接用于Ｔ细胞亚群及其细胞因子的检测。Using directly labelled anti cytokine antibodies with the combination of a fixation and permeabilization method,we have developed the technique for measuring intracellular cytokines by 3 color flow cytometry to test the optimal doses for stimulants and to select the proper time of cell culturing The IFN γ and IL 4 producing CD4 + and CD8 + cells were examined in 5 normal donors The results showed that there was a greater proportion of IFN γ producing CD8 + cells than CD4 + cells,and there were few CD4 + or CD8 + cells producing both IFN γ and IL 4 This method could allow the determinations of cytokine production and the phenotype analysis of the producer cells at single cell level This method can allow the determination of cytokine production as well as the phenotype of the producer cells at the level of individual cells

关 键 词：流式细胞术 细胞因子 三色法 检测 

分 类 号：R446.61[医药卫生—诊断学]