慢性髓性白血病来源的树突状细胞的功能研究

The Function of Dendritic Cells Derived from Chronic Myeloid Leukemia

为研究慢性髓性白血病 (CML)来源的树突状细胞 (dendriticcell,DC)的功能 ,将CML骨髓和外周血单个核细胞在含GM CSF ,IL 4 ,TNF α的培养基中培养 7- 14天。利用流式细胞术对培养细胞进行表型鉴定 ,通过FITC DX(FITC标记的葡聚糖 )摄入实验 ,3H TdR掺入和MTT实验 ,以及LDH释放试验 ,分别检测不成熟DC对外源抗原的摄入能力 ,成熟DC对外源和内源抗原的呈递能力 ,以及细胞毒性T淋巴细胞 (CTL)对白血病细胞的特异杀伤作用。结果表明 :从CML患者骨髓和外周血中能培养出高表达CD1a ,CD86,CD80 ,HLA DR ,CD5 4和CD4的树突状细胞 ;早期DC对FITC DX有吞噬功能 ;培养至出现典型树突状结构的DC(约 7- 10天 )对异体淋巴细胞 (allo MLR)具有较强的刺激作用 ,对自身T细胞也有刺激作用 (auto MLR) ,但相对较弱 ;CML DC能诱导出对自身CML细胞有特异性杀伤作用的CTL。结论提示 ,CML

The purpose of this study was to investigate the function of dendritic cells derived from chronic myeloid leukemia (CML-DC). Mononuclear cells were prepared from bone marrow and peripheral blood of 24 patients with CML, and the DCs were obtained by incubation of MNCs with media containing GM-CSF, IL-4 and TNF-α. The phenotype of CML-DCs was identified by flow cytometry. FITC-dextran uptake, 3H-TdR incorporation or MTT assay and lactate dehydrogenase release assay were used to detect uptake of exogenous antigen in immature DCs, the antigen presenting ability in mature DCs and specific cytotoxicity of CTL to leukemic cells, respectively. The DCs with high expression of CD1a, CD86, CD80, HLA-DR, CD54 and CD4 were obtained from marrow and blood of patients with CML. The uptake of FITC-DX was observed in early DCs. There was a potent stimulation to allo-MLR in DCs cultured for 7-10 days, and a lightly lower stimulation to auto-MLR. CML-DCs can induce the generation of specific cytotoxic T cells. These results suggest that CML-DCs are functional DCs with the ability to induce anti-leukemia effect.