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目的 :探讨利用一定组合的细胞因子 ,将不同亚型的急性髓性白血病 (acute m yeloidleukemia,AML)细胞在体外转化为树突状细胞 (dendritic cell,DC)。方法 :8例初发 AML 患者 (其中M1 1例、M2 a3例、M32例、M4 c1例、M51例 )的骨髓单个核细胞 (BMMNC) ,在含 GM- CSF、IL- 4和TNF-α的完全培养液中培养 14天 ,通过光镜及电镜下观察细胞形态、流式细胞仪测定细胞免疫表型和 MTT法混和淋巴细胞反应来鉴定 DC。结果 :6例患者的 BMMNC培养 14天后转化为 DC。 DC的免疫表型 CD1 a、CD83、CD86 、HL A- DR较培养前明显上调 (P<0 .0 5 )。 DC对同种异体 T淋巴细胞有较强的刺激作用 ,远高于对照 (P<0 .0 5 ) ,而且随 DC数量的增加刺激 T细胞增生的活性越强。2例患者(M2 a1例 ,M31例 )的 BMMNC未转化为 DC,免疫表型分析显示 CD83及 CDla的表达较低 ,但 CD86 及HL A- DR的表达明显增高。结论 :利用一定组合的细胞因子可以将 AML细胞在体外转化为形态、表型和功能符合 DC特征的细胞。某些 AML患者的原始细胞不能转化为 DC,与其亚型无关 ,原因有待进一步探讨
OBJECTIVE: To explore the use of cytokines in combination to transform dendritic cells (DCs) of acute myeloid leukemia (AML) with different subtypes in vitro. Methods: Bone marrow mononuclear cells (BMMNC) from 8 patients with initial AML (M1 1, M2 a3, M32, M4 c1 and M51) were cultured in DMEM containing GM-CSF, IL-4 and TNF- Culture medium for 14 days. The morphology of cells was observed under light microscope and electron microscope, and the immunophenotype of cells was analyzed by flow cytometry and mixed lymphocyte reaction by MTT to identify DCs. RESULTS: BMMNCs from 6 patients converted to DC after 14 days of culture. The immunophenotypes of CD1 a, CD83, CD86 and HL-DR in DCs were significantly up-regulated compared with those before culture (P <0.05). DC had a stronger stimulating effect on allogeneic T lymphocytes, which was much higher than that of the control (P <0.05). Moreover, DCs stimulated the proliferation of T cells with the increase of DCs. BMMNC was not transformed into DC in 2 patients (M2a1, M31). The immunophenotypic analysis showed that the expression of CD83 and CDla was lower, but the expression of CD86 and HL-DR was significantly increased. Conclusion: A certain combination of cytokines can be used to transform AML cells into cells with morphological, phenotypic and DC characteristics. Some AML patients with primitive cells can not be converted to DC, has nothing to do with its subtypes, the reasons to be further explored