用GM-CSF和IL-4在体外诱导高纯度CD14^+树突状细胞

Generation of CD14^+ Dendritic Cells in Vitro with GM-CSF and IL-4

本研究改进传统的树突状细胞 (DC)诱导方法 ,用 GM- CSF(15 0 ng/ml)和 IL- 4 (80 0 U/ml) ,在体外经7d从健康人外周血中诱导出了大量高纯度 DC,其高表达 HL A- 、HL A- 类分子 ,共刺激分子和黏附分子 ,同时还高表达其前体单核细胞的特异性标志 CD14分子 ,显示出成熟 DC的特征。这些 CD14 + DC能强烈诱导同种异体淋巴细胞的增埴 ,其内吞能力在第 3天达最高 ,之后明显下降。此结果丰富了 DC的类型 ,并为 CD14 +

This study was conducted to get high quality and sufficient numbers of mature dendritic cells from healthy donor peripheral blood. The experiment began on culturing of plastic adherent monocytes isolated from healthy donor peripheral blood with granulocyte monocyte clony stimulating factor ( GM CSF 150ng /ml ) and interleukin 4 ( IL 4 800 U/ml ) without fresh medium feeding and cytokines for 7 days. After 7 days, CD14 + monocytes not only differentiated into high purity DC but also expressed HLA Ⅰand HLA Ⅱmolecules , costimulating molecules , adherent molecules and its progenitor marker CD14 molecule highly.These cells displayed all phenotypic and morphologic characteristics of mature dendritic cells and were most potent stimulatory cells in allogeneic mixed leukocyte reactions. The endocytosis ability of these DCs peaked at the third day in culture and decreased remarkably afterwards.These results provide evidence for the first time that CD14 + monocytes differentiated in vitro from peripheral blood monocytes exhibit dendritic cells characteristics and still express its progenitor marker CD14 molecules highly .The results of this experiments may facilitate further studies of CD14 + DC and its clinical applications.