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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 To observe the effect of interleukin (IL) 10 on the dendritic cell (DC) phenotype in vitro and to explore the method and significance of the detection of cell surface antigens by flow cytometry. Methods DCs derived from cord blood CD34 + hematopoietic stem cells were induced by SCF, GMCSF, TGFβ1, Flt3 and TNFα in vitro. DCs were induced by recombinant human IL10 during maturation. The cell phenotypes CD1a, CD11c, CD83, CD80, CD86 and HLADR were analyzed by flow cytometry with direct detection of the three-color fluorescent labeling (FITC, PE, CY) monoclonal antibodies. Results IL10 down-regulated the expression of CD11c, CD83, CD80 and CD86 on the DC surface. Conclusions IL10 can regulate antigen presenting function of DC by inhibiting the expression of co-stimulatory molecules on DC surface. In addition, using trichrome fluorescent labeling assay by flow cytometry can not only save the amount of DC cells, but also have the advantages of rapid and accurate , It is worth promoting application.