临床注射用α-干扰素和粒巨-集落刺激因子诱导慢性髓性白血病细胞向树突状细胞分化  

Differentiation of chronic myeloid leukemia cells to dendritic cells induced by interferon α and granulocyte-macrophage colony-stimulating factor

作　　者：翁开枝[1] 谢晓宝[1] 邱国强[1] 吴浩清[1] 

机构地区：[1]苏州大学附属第三医院血液实验室,常州213003

出　　处：《肿瘤》2007年第1期55-58,共4页Tumor

摘　　要：目的：研究临床注射用α-干扰素（interferon-alpha，IFN-α）和粒巨-集落刺激因子（granulocyte-macrophage colony-stimularing factor，GM-CSF）在诱导慢性髓性白血病（chronic myeloid leukemia，CML）细胞向树突状细胞（dendritic cells，DCs）分化过程中的影响，以建立临床应用稳定的培养体系。方法：取6例CML慢性期患者的骨髓单个核细胞（bone marrow mononuclear cells，BMMNCs），用含10％人AB血清的RPMI 1640同时加入不同细胞因子组合（A组：GM-CSF＋IL-4；B组：临床注射用IFN—α＋临床注射用GM—CSF）作为培养体系，培养7～9d后，在倒置显微镜下观察细胞形态，全自动血液分析仪分析CML-DCs数量，通过流式细胞仪检测其表面标志，采用异基因混合淋巴细胞反应（allogeneic mixed lymphocyte reaction，allo—MLR）分析检测DCs的免疫刺激功能。结果：经不同细胞因子组合分别培养诱导5d和7～9d后的细胞，其特征性表面标志（CD80、CD86、HLA-DR、CD83、CD1a）表达率均高于培养前的细胞（P〈0．05）；在培养的第5天，B组细胞表面标记CD83的表达率高于A组（PG0．05）；但培养7～9d后两组表面标记的表达率无明显差异，CML-DCs计数值也无统计学意义。allo-MLR在DC细胞与反应细胞之比为1：10时B组刺激指数高于A组（P〈0．05）。结论：CML患者的BMMNCs在体外经临床注射试剂组合培养后可以分化为具有典型形态、免疫表型和一定功能的DCs，这一结果为培养更适合回输人体的“临床型”DCs提供了新的培养体系。Objective： To investigate the influence of interferon-alpha （IFNα）and granuloeyte-maerophage colony-stimulating factor （GM-CSF） on the differentiation of chronic myeloid leukemia （CML） cells to dendritic cells （DCs） in vitro and establish a stable cell culture system for clinical use. Methods： Bone marrow mononuclear cells （BMMNCs） were isolated from 6 patients at the chronic phage of CML. CML-DCs were cultured in RPMI-1640 supplemented with 10% human AB serum in the presence of different cytokines for 7-9 d. Group A： GM-CSF q-interleukin （IL）-4; Group B： IFNa for clinical injection ＋GM-CSF for clinical injection. The morphologic features of CML-DCs were observed by inverted microscope and the number of CML-DCs was counted through full automatic blood analyzer. The immunophenotypes were analyzed by flow cytometry. The immune function of CML-DCs was detected through allogeneic mixed lymphocyte reaction （allo-MLR）. Results： After culture with different cytokines for 5 d and 7-9 d, the immunophenotypes of CML-DCs （CD80, CD86, CD83 and HLA-DR） were significantly up-regulated compared with pre-culture （P〈0.05）. The CD83 expression was higher in Group B than that in group A after cultured for 5 d （P〈0.05）. But the expression of the immunophenotypes were not significantly different between the two groups after cultured for 7-9 d. There was no significant difference in the number of CML-DCs between the two groups. When the ratio of DC ： T was 10 ： 1, the Allo-MLR in group B was significantly higher than that in group A （P〈0.05）. Conclusions： Combination of clinical injections can induce differentiation of BMMNCs isolated from CML patients to DCs which have typical morphology, marked immunophenotypes, and biological function. It provides a new DCs culture system for clinical vaccination in vitro.

关 键 词：白血病 髓性 慢性 Α-干扰素 粒巨-集落刺激因子 树突细胞 

分 类 号：R733.72[医药卫生—肿瘤] R73-36[医药卫生—临床医学]