IFN-γ激活的树突状细胞对患者急性髓性白血病细胞的杀伤作用  

Cytotoxicity of IFN-γ-Activated Dendritic Cells to Freshly Isolated Acute Myeloid Leukemia Cells

作　　者：石军[1] 张毅[2] 苏基莹[1] 李晓[1] 浦权[1] 池田和真[3] 

机构地区：[1]上海交通大学附属第六人民医院血液科,上海200233 [2]上海交通大学附属第六人民医院药剂科,上海200233 [3]日本冈山大学附属医院第二内科

出　　处：《中国实验血液学杂志》2005年第6期1071-1075,共5页Journal of Experimental Hematology

基　　金：上海市青年科技启明星课题(沪科(2002)270号);上海市卫生局青年课题(编号:024Y14)资助项目

摘　　要：为检测干扰素γ(IFNγ)激活的外周血树突状细胞(DC)对患者急性髓细胞白血病(AML)细胞的直接杀伤活性并探讨其杀伤机制,分离健康供者外周血单核细胞,用重组粒巨噬细胞集落刺激因子和白细胞介素4诱导生成DC。于诱导第7天在合成培养液中加入IFNγ继续培养12小时,将其激活。从6例初发的急性髓细胞白血病患者外周血分离单个核细胞作为靶细胞,以不同效靶比与DC共同培养18小时,采用51Cr释放试验检测DC激活前后抗肿瘤活性的差异。同时,用流式细胞术检测DC表面共刺激分子,细胞膜及细胞内Fas配体(FasL)、TNFα及肿瘤坏死因子相关凋亡诱导配体(TRAIL)的改变,以明确IFNγ激活的DC对肿瘤细胞直接杀伤活性的作用机制。结果表明:IFNγ可增强DC对AML细胞的杀伤活性,在效靶比为20∶1时杀伤率为(33.8±1.6)%,与未加刺激因子前相比有显著差异(P<0.05);IFNγ可上调DC表面共刺激分子CD86和CD83的表达;IFNγ刺激后,DC细胞内TRAIL表达明显增强,但其在细胞表面仍呈现低表达,而细胞内及细胞表面FasL及TNFα在IFNγ刺激前后均无明显变化。结论:IFNγ激活的DC可有效杀伤患者AML细胞,其作用机制与DC的成熟及TRAIL凋亡途径部分相关,而与FasL及TNFα凋亡途径无关。To investigate the tumoricidal activity of dendritic cell （DC） stimulated by interferon-γ（IFN-γ） against freshly isolated myeloid leukemia cells and its mechanism, the peripheral blood monocytes collected from healthy donors were cocultured with interleukin-4 and granulocyte-machrophage colony- stimulating factor in medium to induce DC for 7 days, After 12 hour culture in the absence or presence of IFN-γ, the changes of costimulatory molecules were analyzed with flow cytometry. To assay the cytotoxicity of DC against freshly isolated acute myeloid cells, they were cocultured at various effector-to-target ratio for 18 hours, then the percentage of tumoricidal activity was measured with ^51Cr release assay, To explore the mechanism of DC-mediated cytotoxicity, the change of DC surface or intracellular protein expression of Fas ligand （Fas L）, TNF-α and TNF related apoptosis-inducing ligand（TRAIL） were analyzed with flow cytometry. The results showed that IFN-γ enhanced cytotoxicity of DC against AML cells was （33.8 ± 1.6 ）% at E： T as 20： 1, compared with unstimulated DC （P 〈 0.05 ） ; IFN-γ up-regulated expression of costimulatory molecules of DC surface such as CD86 and CD83; after stimulation with IFN-γ, expression of intracellular TRAIL of DC was significantly enhanced, but expression of TRAIL on cell surface of DC was low; while the significant changes of Fas L and TNF-α expression neither on cell surface or in cells were not observed before or after stimulation with IFN-γ. It is concluded that DC stimulated by IFN-γ exhibit tumoricidal activity against AML cells, The cytotoxicity is partially related to maturation of DC and TRAIL inducing apoptosis, but not associated with death domain-independent mechanism of Fas L and TNF-α.

关 键 词：IFN-Γ 树突状细胞 急性髓细胞白血病 肿瘤杀伤活性 

分 类 号：R733.7[医药卫生—肿瘤]