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目的:探讨钙离子载体(calcium ionophore,CI)对外周血单核细胞来源的树突状细胞(DC)的影响。方法:分离健康献血者外周血单核细胞,分别加入重组人粒/单集落刺激因子(rhGM-CSF)100μg/L、rhGM-CSF 100μg/L+CI 10μg/L及rhGM-CSF+CI各100μg/L。体外培养40 h后,于光镜及电镜下观察细胞的形态、流式细胞仪检测细胞的表面标志,MTT比色法检测上述分子对自体T细胞的刺激增殖作用。结果:外周血单核细胞在GM-CSF+CI各100μg/L的条件下培养40 h,就可看到典型的DC形态,其表面CD14分子表达减少,HLA- DR、CD40、CD83及CD86分子的表达明显增高,且具有明显 刺激自体T细胞增殖的能力。结论:CI有显著加速GM-CSF 诱导的外周血单核细胞向DC转化的作用。
Objective: To investigate the effects of calcium ionophore (CI) on dendritic cells (DCs) derived from peripheral blood mononuclear cells. Methods: Peripheral blood mononuclear cells from healthy blood donors were isolated and treated with rhGM-CSF 100μg / L, rhGM-CSF 100μg / L + CI 10μg / L and rhGM-CSF + / L. After cultured in vitro for 40 h, the morphology of cells was observed under light microscope and electron microscope. The surface markers of cells were detected by flow cytometry. The proliferation of autologous T cells was detected by MTT assay. Results: After being cultured for 40 h in the presence of GM-CSF + CI 100 μg / L, peripheral blood mononuclear cells showed a typical DC morphology with reduced expression of CD14 molecules on the surface. HLA-DR, CD40, CD83 and CD86 molecules The expression was significantly increased, and has the ability to significantly stimulate autologous T cell proliferation. Conclusion: CI can significantly accelerate the GM-CSF-induced peripheral blood mononuclear cells to DC transformation.