干扰素-α联合GM－CSF诱导慢性髓性白血病细胞向树突状细胞分化

目的：研究干扰素-α（IFN-α）对慢性髓性白血病（CML）骨髓单个核细胞来源的树突状细胞（DCs）发育的影响。 方法： 12例初发慢性期CML患者的骨髓单个核细胞，分别与含如下细胞因子,RPMI-1640培养液共育：rhGM-CSF 1×106U/L联合rhIFN-α 2×106U/L（IFN-α组）、rhGM-CSF 1×10⁶U/L联合rhIL-4 5×10⁵U/L（IL-4组）、单用rhGM-CSF 1×10⁶U/L和单用rhIFN-α 2×106U/L，培养7 d；于第8-10 d，部分孔加入rhTNF-α 5×10⁴U/L。形态学（Wright染色、倒置显微镜）、免疫学（CD80、CD86、CD83、CD1a、HLA-DR）检测；磷脂酰丝氨酸（PS）转位检测 DCs凋亡情况；荧光原位杂交（FISH）对1例CML进行细胞遗传学分析；混合淋巴细胞反应（MLR）检测刺激同种异体T淋巴细胞增殖的能力。 结果： CML骨髓单个核细胞经上述细胞因子诱导7 d后，IFN-α组和IL-4组均呈现树突状细胞的典型形态；免疫学鉴定，IFN-α组DCs CD80、CD86、CD83、HLA-DR的表达显著高于IL-4组（P<0.05），经5×104U/L rhTNF-α作用后，两组DCs CD80、CD86、CD83、HLA-DR进一步上调，其中IFN-α组DCs CD80、CD86、CD83、HLA-DR的表达显著高于IL-4组（P<0.05）；经FISH证实来源于白血病细胞；两组DCs均具有刺激同种异体T淋巴细胞增殖的能力，IFN-α组刺激淋巴细胞增殖的能力明显高于IL-4组(P<0.05)。 结论： IFN-α可促进CML骨髓单个核细胞来源的树突状细胞的分化、活化。这可能是IFN-α在CML中的治疗机制之一。

Induction of dendritic cells with IFN-alpha and GM-CSF from bone marrow mononuclear cells from patients with chronic myeloid leukemia

AIM: To investigate the effects of interferon-alpha (IFN-α) on the development of dendritic cells (DCs) derived from bone marrow mononuclear cells in patients with chronic myeloid leukemia (CML). METHODS: Bone marrow mononuclear cells from 12 CML patients were cultured initially using cytokines as follows: recombined human granulocyte-macrophage colony stimulating factor (rhGM-CSF) plus IFN-α (IFN-α-DCs); rhGM-CSF plus recombined human interleukin-4 (rhIL-4) (IL-4-DCs); IFN-α alone; rhGM-CSF alone in 10% FBS RPMI-1640 medium for 7 days and then recombined human tumor necrosis factor-α (rhTNF-α) was added for another 3 days. The morphologic features were observed by Wright's staining under inverted microscope. CD80，CD86，CD83，CD1a and HLA-DR expression were assayed by flow cytometry. Cytogenetic analysis was performed for one CML patient by fluorescence in-situ hybridization (FISH), and the functions of antigen presenting were tested by mixed lymphocyte reaction (MLR). RESULTS: IFN-α-DCs displayed features in morphology that was similar to those of IL-4-DCs with delicate membrane projections. IFN-α-DCs showed an increase in expression of CD80, CD86, CD83, HLA-DR and more intense abilities of allogeneic antigen presentation with and without rhTNF-α stimulation, compared with the control groups of IL-4-DCs. FISH confirmed the DCs of both groups were leukemic origin. CONCLUSIONS: (1) IFN-α promoted the differentiation/activation of bone marrow mononuclear cells from patients with CML into activated dendritic cells. (2) The phenomenon of generation of activated DCs in vitro might contribute to therapeutic effect of IFN-α in CML. (3) IFN-α may be valuable for the generation of active bone marrow mononuclear cells-derived DCs to be as vaccination strategies of CML patients.