组合性细胞因子与钙离子载体诱导急性髓系白血病细胞向树突状细胞分化中的作用

Effects of cytokine combinations and calcium ionophore on differentiation of dendritic cells from acute myeloid leukemia cells

目的:探讨钙离子载体能否诱导急性髓系白血病细胞分化为树突状细胞及其抗肿瘤免疫。方法:选择2004-01/09解放军广州军区广州总医院血液科住院的初诊或复发的急性髓系白血病患者7例。分离急性髓系白血病患者的急性髓系白血病细胞,在体外给予钙离子载体100μg/L培养3～5d(钙离子载体组),或1000U/mL重组人粒细胞-巨噬细胞集落刺激因子、1000U/mL白细胞介素4和500U/mL肿瘤坏死因子α培养7～10d(组合细胞因子组)。通过细胞形态的观察及细胞免疫表型的检测鉴定树突状细胞,用同种异体混合淋巴细胞实验及细胞毒实验检测树突状细胞的功能。结果:①形态学观察和免疫表型变化:钙离子载体组及组合细胞因子组诱导急性髓系白血病细胞均可获得具有典型树突状细胞形态及免疫表型的成熟树突状细胞,但钙离子载体组诱导的树突状细胞表面CD80,CD86,CD83,CD40,CD54等分子的表达较组合细胞因子组明显增高。②混合淋巴细胞反应和胞毒性T淋巴细胞活性:钙离子载体组所诱导的急性髓系白血病细胞刺激同种异体混合淋巴细胞反应的能力及激发细胞毒性T淋巴细胞的活力比组合细胞因子组强。结论:钙离子载体能高效诱导急性髓系白血病细胞分化成树突状细胞,比组合细胞因子诱导分化的树突状细胞功能更强。

AIM： To explore the feasibility of dendritic cells （DC） differentiate from acute myeloid leukemia （AML） cells with induction of calcium ionophore （CI） and their anti-tumor immunity. METHODS： Seven inpatients of first episode or recurrent AML were selected from the Department of Hacmatology, General Hospital of Guangzhou Military Area Command of Chinese PLA from January to September 2004. AML cells were isolated and cultured in vitro in the presence of 100 μg/L CI for 3-5 days （CI group） or in the presence of 1 000 U/mL granulocyte macrophage colony stimulating factor （GM-CSF）, 1 000 U/mL interleukin 4 （IL-4） and 500 U/mL tumor necrosis factor alpha （TNF-α） for 7-10 days （cytokine combination, CK group）. DC was evaluated with morphologic observation and detection of cellular immunological phenotype. And functions of DC were examined by mixed lymphocyte reaction （MLR） and cytotoxicity test. RESULTS： （1）Morphologic observation and immunological phenotype： After cultured with CI or cytokine combinations, all the AML cells from 7 patients developed into mature DC of morphologic and phenotypic characters. The expressions of CD40, CD54, CDS0, CD86 and CD83 molecules on surface of induced DC in CI group were significantly increased com- pared with CK greup.（2）MLR and cytotoxicity： CI-induced AML cells pos- sessed enhanced ability of stimulating MLR and accelerating cytotoxicity T lymphocyte compared with CK-induced cells. CONCLUSION： DC can differentiate from AML cells with highly efficient induction of CI, and the functions of CI-induced DC are enhanced notably compared with CK-induced.