细胞因子体外诱导急性髓系白血病细胞分化为树突状细胞  

Dendritic cells (DC) induced from acute myeloid leukemia (AML) cells with cytokine cocktails

作　　者：严匡华[1] 尤胜国[1] 卞寿庚[1] 马冠杰[1] 葛薇[1] 马双 刘世和[1] 赵春华[1] 

机构地区：[1]中国医学科学院,中国协和医科大学血液学研究所,血液病医院,天津300020

出　　处：《中华血液学杂志》2003年第7期365-368,共4页Chinese Journal of Hematology

基　　金：卫生部专项基金资助项目 (WKZ 2 0 0 0 1 3 4)

摘　　要：目的　研究细胞因子组合体外诱导急性髓系白血病 (AML)细胞分化为树突状细胞 (DC)的可行性及AML细胞衍生DC(AML DC)的生物学特性。方法　AML细胞分别在GM CSF +IL 4、GM CSF +TNF α或GM CSF +IL 4 +TNF α 3种细胞因子组合以及不含细胞因子的培养液中培养。通过细胞形态动态观察、细胞化学染色和细胞免疫表型鉴定DC。用混合淋巴细胞培养 (MLC)、FITC标记的葡聚糖摄入实验和LDH释放实验检测DC功能。RT PCR和FISH检测AML DC的特异融合基因。结果　15例AML细胞在 3种细胞因子作用下发生典型的DC形态变化。AML DC的DC相关表面分子CD1a、CD80 、CD86 、CD1 0 6 、CD83和HLA DR等较未培养或不加细胞因子培养的AML细胞表达明显上调 (P <0 0 5 )。AML DC的异基因刺激能力明显高于未培养或不加细胞因子培养的AML细胞 (P <0 .0 5 )。只有用GM CSF +IL 4培养的AML DC有吞噬能力。AML DC与未培养AML细胞致敏的疾病初发时分离的T细胞比较 ,对自体AML细胞无明显的杀伤活性。AML DC仍具有未培养AML细胞的特异融合基因。结论　体外细胞因子可诱导各型AML细胞分化为DC ,细胞因子组合不同 ,AML DC的成熟状态可有一定的差异。AML DC不但起源于AML细胞 ,而且具有正常DC的典型形态、表型和功能。Objectives To explore the feasibility of DC being in vitro induced from AML cells with cytokine cocktails and their biological properties. Methods AML cells were cultured in either presence or absence of cytokine cocktails. DC were studied for morphology, and cytochemical and immunofluorescent staining. Functions of DC were examined by MLC, FITC conjugated dextran uptake test, and LDH release assay. RT PCR and FISH were used to analyze the specific fusion genes of culture derived DC. Results Classical DC morphological changes occurred in all 15 cultured AML cells. DC associated surface molecules such as CD 1a , CD 80 , CD 86 , CD 106 , CD 83 and HLA DR were upregulated (P<0.05). The allostimulatory abilities of culture derived DC were significantly higher than those of AML cells uncultured or cultured in the absence of cytokines (P<0.05). Culture derived DC only in the presence of GM CSF+IL 4 have phagocytotic activities. CTL assay was performed in 5 of the 15 samples. At effector/target ratio of 20∶1, auto T lymphocytes primed with the culture derived DC exhibited no more killing activity to auto AML cells than those stimulated by IL 2 or uncultured AML cells. Culture derived DC presenced the native AML specific aberrant karyotype and related fusion gene. Conclusions Cytokine cocktails could in vitro induce AML cells into DC with classical morphology, immunophenotype and function. DC maturity induced by different cytokine cocktails could be variable. Culture derived DC were originated from the native AML cells. AML cells could make the auto T lymphocyte anergy.

关 键 词：急性髓系白血病 细胞分化 树突状细胞 粒-巨噬细胞集落刺激因子 白细胞介素-4 肿瘤坏死因子-α 

分 类 号：R733.71[医药卫生—肿瘤]