钙离子载体联合GM．CSF体外诱导外周血单核细胞生成树突状细胞

目的：利用新型诱导剂钙离子载体A23187联合重组人粒细胞-巨噬细胞集落刺激因子（rhGM-CSF）体外诱导人外周血单核细胞（PBMC）生成树突状细胞（DCs）。方法：采用密度梯度离心法及黏附法分离出PBMC，按不同培养方法分成：传统方法组，rhGM-CSF＋重组人白介素-4（rhIL-4）＋重组人肿瘤坏死因子-α（rhTNF-α）；新型诱导剂组，加入rhGM—CSF4＋A23187。光镜观察细胞形态的变化，流式细胞仪检测DC细胞的表面标志，MTT比色法检测各组DCs刺激同种异体外周血T细胞增殖的能力。结果：与传统方法相比，A23187联合rhGM—CSF诱导培养的DCs更具有典型的树突形态；DC表面分子CD83D1a、CD86、CIM0表达（45．2％、31．5％、40．1％、36．4％）较传统方法组（16．9％、20．4％、26．5％、22．3％）明显上调（P〈0．05），而CD14表达（5．7％vs19．0％）明显下降（P〈0．05），刺激同种异体T细胞增殖的能力更强。结论：新型诱导剂钙离子载体A23187联合GM-CSF能更有效地诱导PBMC生成强效成熟的DCs。

Cultivation of dendritic cells from human peripheral blood monocytes cells induced by calcium ionophore associated with GM-CSF in vitro

To evaluate the effect of calcium ionophore （CI） A23187 associated with human recombinant granulocyte-macrophage colony stimulating factor （rhGM-CSF） on the cultivation of dendritic cells （DCs） from healthy human peripheral blood monocytes cells （PBMCs） induced in vitro. Methods： PBMCs isolated with density gradient centrifugation and adhesion were divided into two groups according to different cultural methods ： traditional control group with rhGM-CSF ＋ rhIL-4 ＋ rhTNF-o~ added and novel inductive group with rhGM-CSF ＋ CI A23187 added. The morphology of the DCs was observed by microscope, the surface marker monitored by flow cytometry, and multiplication capacity of DCs-stimulated T-cells in allogenic peripheral blood detected with MTT assay. Results：Compared with traditional method, more typical morphological features could be observed in DCs induced by CT A23187 associated with rhGM-CSF. The expressions of DC phenotype molecules including CD83, CDla, CD86 and CIM0 in inductive group （45.2% , 31.5% , 40.1% , 36.4% ） were increased more significantly than those in traditional control group （ 16.9% , 20.4% , 26.5% , 22.3% ） , respectively （P〈0.05）, while the expression of CD14 was remarkably decreased （5.7% vs 19.0% ） （P 〈 0. 05）. Moreover, the multiplication capacity of DCs-stimulated T-cells was greatly enhanced. Conclusion： CI A23187 associated with GM-CSF could be more effective in inducing PBMCs into powerful mature DCs.