人外周血树突状细胞培养和地塞米松对其分化的影响作用

Culture and comparison of the morphology and function of dendritic cells from human peripheral blood monocytes with and without dexamethasone

目的分离培养和鉴定人外周血树突状细胞(DC),以及探讨地塞米松对其分化的影响作用。方法密度梯度离心法分离人外周血单个核细胞,贴壁后加入GM-CSF、IL-4和LPS培养,部分组另加入地塞米松,观察细胞形态学、流式标志和DC与T淋巴细胞共培养后的增殖变化。结果外周血单核细胞诱导培养后具有DC形态学特征,CD83表达上调,CD14表达下调,DC与T淋巴细胞共培养后呈增殖反应。培养液中加入地塞米松后CD83表达下调,CD14表达上调,DC与T淋巴细胞共培养后增殖反应减弱。结论外周血单核细胞经联合细胞因子可诱导为DC;地塞米松可使DC在功能上处于不成熟状态。

Objective： To study the morphology and function of the dendritic cells （DC） from human peripheral blood monocytes and the change in cell differentiation and maturation after DC treated with dexamethasone （Dex-DC）. Methods： DCs were sorted from human monocytes by culturing in presence of cytokines （ GM-CSF, IL-4 ） and LPS for 8 days with or without dexamethasone. The change in morphology, function and phenotypic characterization was compared. Results：The cells treated with or without Dex, developed a characteristic dendritic morphology, however, Dex-DC expressed higher level of CD14 and lower level of CD83 than the untreated cells. In addition, a low APC function was demonstrated by Dex-DC. Conclusion： DCs can be induced from peripheral blood monocytes in medium with cytokines ; the cultured PS in presence of Dex are shown at a more immature stage, indicating that Dex modulates DC differentiation, maturation, and function.