流式细胞仪三标记法检测白细胞介素-10对人树突状细胞表型的影响  

Effect of Interleukin-10 on the Phenotype Cultured Human Dendritic Cells by Flow Cytometry of Three Fluorescence Marker

作　　者：史桂英[1] 孙桂芝[1] 周同[2] 金慧芳[2] 张玉梅[2] 赵亚鹏[2] 陈楠[2] 

机构地区：[1]第二医科大学细胞生物学教研室,上海200025 [2]上海第二医科大学瑞金医院肾内科

出　　处：《中国微循环》2005年第3期185-188,F002,共5页Journal of Chinese Microcirculation

基　　金：国家自然科学基金(39970340);上海市自然科学基金(02ZB14041;034119916)

摘　　要：目的观察白细胞介素(IL)10对体外培养人树突状细胞(DC)表型的影响,探讨流式细胞术三色荧光标记抗体检测细胞表面抗原的方法及意义。方法通过SCF、GMCSF、TGFβ1、Flt3和TNFα体外培养体系,将脐血CD34+造血干细胞诱导扩增获得DC,并于成熟过程中用重组人IL10进行干预。采用流式细胞术的三色荧光标记(FITC、PE、CY)单克隆抗体直接检测技术,分析细胞表型CD1a、CD11c、CD83、CD80、CD86和HLADR。结果IL10可下调成熟中DC表面CD11c、CD83、CD80和CD86的表达。结论IL10通过抑制DC表面黏附共刺激分子表达,可调节DC的递呈抗原功能;此外,采用流式细胞仪的三色荧光标记检测方法,不仅可以节约DC细胞用量,而且具有快速和准确的优点,值得推广应用。Objective The present study was designed to detect the influence of interleukin (IL)-10 upon the morphologic features of cultured human dendritic cells (DCs), to investigate the method and the meaning of phenotype examined by flow cytometry of three fluorescence marker. Methods Cord blood CD34 + stem cells were isolated and cultured in IMDM medium with SCF、GM-CSF、TGF-β_ 1 、Flt-3、TNF-α. DCs were interfered by recombination of human IL-10. HLA-DR、CD1a、CD11c、CD83、CD80 and CD86 were assayed by flow cytometry of three fluorescence marker(FITC-、PE-、CY-).Results The expression of CD11c、CD83、CD80 and CD86 was lower in IL-10 group. Conclusions IL-10 might inhibit the expression of adhesion and co-stimulation molecules. It was presumed that IL-10 might adjust the antigen presenting function of DCs. In addition, by using the method of three fluorescence marker, we could save DC number needed. Moreover this method is fast and exact, which is deserved to be applied.

关 键 词：流式细胞术 树突状细胞 细胞表型 白细胞介素-10 

分 类 号：Q25[生物学—细胞生物学]