【摘要】 目的探讨从脐血CD34+造血细胞诱导DC2的方法及CD40分子在DC2分化诱导中的作用。方法运用磁珠从脐血中分离出CD34+造血干细胞,以rhIL3(10ngmL)、rhFlt3L(100ngmL)和rhGMCSF(100ngmL)诱导其向DC2分化,采用流式细胞仪分析DC2表型,并观察抗人CD40单克隆抗体诱导DC2分化成熟的作用。结果人脐血CD34+造血细胞在rhIL3、rhFlt3L和rhGMCSF联合诱导培养12d,获得具有DC2样(淋巴样DC)形态的细胞,然后分别加入rhIL3+抗人CD40mAb(Ⅰ组)或rhIL3+TNFα(Ⅱ组)诱导DC2的分化成熟,流式细胞仪分析细胞表型,发现Ⅰ组和Ⅱ组Lineage-(CD3、CD14、CD16、CD19、CD56)CD123+细胞的HLADR、CD83、CD86、CD80表达率分别为88.78%、37.38%、32.83%和99.08%;78.87%、32.29%、29.57%和98.86%。结论体外联合多种细胞因子从CD34+造血细胞成功地诱导出富集DC2表型的细胞,抗人CD40mAb可以促进DC2的分化成熟。更多还原

【Abstract】 Objective To develop a method of inducing DC2 from human cord blood CD34~+ hemopoietic cells and evaluate the effect of CD40 signal on differentiation and induction of DC2. Methods CD34~+ hemopoietic cells were isolated from cord blood of healthy volunteers with CD34-FITC and anti-FITC labeled magnetic microbeads, then cultured with rhIL-3 (10 ng/mL), rhFlt-3L (100 ng/mL), and rhGM-CSF (100 ng/mL). The surface molecules of DC2 were identified by FCM and the role of anti-CD40 mAb in differentiation and maturation of DC2 was analyzed by cytokines assay. Results CD34~+ hemopoietic cells of human cord blood were cultured with rhIL-3, rhFlt-3L, and rhGM-CSF for 12 days, and then rhIL-3+anti-human CD40mAb (groupⅠ) or rhIL-3+ TNF-α (groupⅡ) were added respectively to induce their differentiation and maturation. The expression rates of surface molecules HLA-DR, CD83, CD86, and CD80 on Lineage~-CD123~+ DC2 induced by groupⅠwere 88.78%, 37.38%, 32.83%, and 99.08%, respectively; otherwise the same surface molecules on Lineage~-CD123~+ DC2 induced by groupⅡ were 78.87%, 32.29%, 29.57%, and 98.86%, respectively. Conclusion DC2 can be induced from cord blood CD34~+ hemopoietic cells with cytokines cocktail in vitro. The agonist anti-human CD40mAb can enhance the differentiation and maturation of DC2.更多还原