慢性粒细胞白血病树突状细胞的体外定向诱导扩增及鉴定

Generation of dendritic cells from peripheral blood of patients with chronic myelogenous leukemia

目的 体外定向诱导扩增慢性粒细胞白血病 (CML)患者外周血单个核细胞 (PBMNC)来源的树突状细胞 (CML -DC)并测定其表型。方法 取CML患者新鲜抗凝血 ,用淋巴细胞分离液分离PBMNC ,经细胞因子rhGM-CSF、rhIL - 4、rhTNF -α联合刺激培养 9～ 12d ,收集培养细胞 ,光镜形态学分析并细胞计数 ,结合PE -CD1a单抗、FITC -CD14单抗在流式细胞仪 (FCM)上进行CML -DC比例及表型分析。结果 CML患者PBMNC经以上细胞因子联合培养 ,CML -DC含量可达 3.5 %～ 2 3.5 % ,而新鲜分离的CML患者PBMNC中CML -DC含量仅为0 .1%～ 1%。结论 CML患者PBMNC经体外定向诱导扩增培养可以生成较多的CML -DC ,从而为CML的免疫治疗应用于临床提供了基础。

Objective To expand dendritic cells(DC) from peripheral blood of patients with chronic myelogenous leukemia(CML) and analyze their cell surface phenotype. Methods Mononuclear cells were isolated from peripheral blood of CML patients and cultured with rhGM-CSF,rhIL-4 and rhTNF-α in vitro for 9～12 d. The morphology of the expanded cells was studied under microscope and the phenotypes were analyzed using a FACScan Flow Cytometer. Results The percentage of CML-DC (expressed as CD1a+ but CD14-) could be increased from 0.1%～1.0% to 3.5%～23.5% by culturing in prensence of rhGM-CSF, rhIL-4 and rhTNF-α. Conclusions Certain amount of CML-DC could be generated from the peripheral blood of CML patients,rhTNF-α by in vitro culture with certain cytokines which provide a basis of clinical immunotherapy of CML-DC.