Serum-free culture of dendritic cells from patients with chronic myeloid leukemia in vitro and estimation of their cytotoxicity  

慢性粒细胞白血病树突状细胞的体外无血清培养及杀瘤效应研究(英文)

作　　者：赵文理[1] 邢佩霓[1] 魏续仓[1] 王彤[1] 杨娣娣[1] 李梅生[1] 

机构地区：[1]陕西省人民医院即西安交通大学陕西医院血液科,西安710068

出　　处：《Chinese Medical Journal》2002年第9期1296-1300,143-144,共5页中华医学杂志（英文版）

基　　金：ThisresearchwassupportedbyagrantfromtheShannxiProvincialScienceFoundationofPublicHealthBureau (No .0 0 12 2 )

摘　　要：OBJECTIVE: To establish a serum-free culture system of dendritic cells (DCs) from chronic myeloid leukemia (CML) cells so that DCs vaccine may be applied to the adoptive immunotherapy of CML in the near future. METHODS: Fetal calf serum, serum-free medium and autologous serum were used for culture of DCs. The usage of cytokines was classified into two groups: group A (stem cell factor, granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4) and group B (granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4). The phenotypes of DCs were analyzed by using indirect immunofluorescence and flow cytometry. Mixed leukocyte responses were performed by methyl thiazolyl tetrazolium (MTT) assay. Chromosome analysis of DCs can be achieved by displaying G banding. T cells from CML patients were stimulated with autologous DCs and T-cell cytotoxicity was measured by (MTT) assay. RESULTS: CD34(+) cells or mononuclear cells were obtained from peripheral blood or bone marrow samples of eight patients of chronic-phase CML. Group A of serum-free medium was better than group B in expansion of total cell numbers and the rate of DCs. These results of serum-free medium were not significantly different from those of fetal calf serum medium, but the results of autologous serum medium were inferior to two groups above. The expression of major histocompatibility complex class II antigen on the surface of DCs was notable (> 50%), but the expression of CD83 and the costimulatory molecules CD86 was not noticeable (10% - 50%). Although CD1a(+)/CD14(-) DCs were potent stimulators of allogeneic lymphocytes, expansion of T cells from normal volunteers were not significant (average 27.2 fold at DCs: T cells ratio of 1:10). At day 12, CD1a(+) cells from three patients were studied by displaying G banding and Ph(+) cells in these populations were 100%, 98% and 60%, respectively. At an effector: target ratio of 40:1, 32% to 45% cytotoxicity was noted with DC-stimulated T目的　拟建立慢性粒细胞白血病 (CML)树突状细胞 (DCs)体外无血清培养体系 ,以期将来用于CML的过继性免疫治疗。方法　小牛血清 (FCS)、无血清 (SFM)及自体血清分别培养CML病人外周血 /骨髓CD34+细胞或单个核细胞 ,以SCF、GM CSF、TNF α和IL 4 (A)与GM CSF、TNF α和IL 4 (B)两组不同的细胞因子作对照。间接免疫荧光法及流式细胞术鉴定CML DCs表型 ,甲基四噻唑蓝 (MTT)法测定CML DCs刺激同种异体T细胞反应能力 ,G显带技术检测培养DCs携带Ph染色体比例 ,MTT法评价CML DCs刺激自身T细胞杀伤自体白血病细胞效应。结果　 8例CML慢性期病例 ,SFM条件下A组培养体系在总细胞扩增倍数及DCs比例两方面均明显优于B组 ,SFM同FCS体系相比无显著差异 ,但自体血清培养结果低于以上两种体系。CML DCsMHC Ⅱ类分子呈高表达(>5 0 % ) ,但CD83和CD86表达比例不高 (10 % - 5 0 % ) ,且刺激同种异体T细胞反应能力不强。 3例患者检测了培养CML DCsPh染色体比例 ,分别为 10 0 %、98%及 6 0 % ,与培养前比例基本吻合 ;同时以CML DCs同自身T细胞、自体白血病细胞共孵育 ,测其杀伤率为 38 5 %± 6 5 % (效靶比为 4 0∶1)。结论　①建立了稳定的无血清培养CML DCs体系 ;②CML DCsMHC Ⅱ类分子显著表达 ,CD83和CD86表达比例不高 ,

关 键 词：Cells  Cultured Culture Media  Serum-Free Cytotoxicity  Immunologic Dendritic Cells Humans Immunotherapy  Adoptive Leukemia  Myeloid  Chronic Research Support  Non-U.S. Gov't T-LYMPHOCYTES 

分 类 号：R733.72[医药卫生—肿瘤]