IFN-α联合GM-CSF诱导胃癌患者外周血单个核细胞分化为树突状细胞

Dendritic cells induced by IFN-α combined with GM-CSF from peripheral blood mononuclear cells of gastric cancer patients

目的:探索干扰素-α(interferon-α,IFN-α)联合粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stim-ulating factor,GM-CSF)体外诱导胃癌患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)向树突状细胞(dendriticcell,DC)分化的可能性。方法:10例胃癌患者PBMC分别用GM-CSF 100 ng/ml联合IFN-α500 IU/ml(命名为IFN-αDC)或GM-CSF 100 ng/ml联合50 ng/ml IL-4(命名为IL-4 DC)体外培养,然后用CD40L、LPS诱导DC成熟。Giemsa染色法观察IFN-αDC和IL-4 DC的形态,流式细胞术分析IFN-αDC和IL-4 DC表面CD1a、CD80、CD83、CD86和HLA-DR的表达情况,同种异体混合淋巴细胞反应(mixed lymphocyte reaction,MLR)检测不同的成熟DC刺激同种异体T淋巴细胞增殖的能力。结果:IFN-αDC和IL-4 DC均呈现典型DC形态。IFN-αDC和IL-4 DC分别在诱导第3天和第5天时,细胞表面CD1a、CD80、CD83、CD86和HLA-DR表达达到较高水平,成熟IFN-αDC表面CD83[(78.25±15.36)%vs(50.14±10.24)%,P<0.05]和CD86[(84.84±10.12)%vs(62.93±15.12)%,P<0.05]的表达均高于成熟IL-4 DC。成熟IFN-αDC刺激异体T淋巴细胞增殖能力强于未成熟IFN-αDC(P<0.05)。在DC与T细胞数量比为1∶40和1∶20时,成熟IFN-αDC刺激同种异体T淋巴细胞增殖的能力明显强于成熟IL-4 DC[(39.43±9.21)%vs(27.34±10.63)%,(60.31±7.86)%vs(48.63±6.25)%;均P<0.05]。结论:相比常用的IL-4联合GM-CSF诱导方法,IFN-α联合GM-CSF可以在更短时间内将胃癌患者PBMC诱导成具有更强刺激同种异体T淋巴细胞增殖能力的DC细胞,这可能与其表面CD83和CD86表达增高有关。

Objective：To investigate the possibility of inducing dendritic cells （DCs） by interferon-α （IFN-α） com- bined with granulocyte-macrophage colony-stimulating factor （GM-CSF） from peripheral blood mononuclear cells （PB- MCs） in gastric cancer patients. Methods： PBMCs from 10 gastric cancer patients were cultivated using granulocyte mac- rophage colony stimulating factor （GM-CSF） 100 ng/ml combined with IFN-ot 500 IU/ml （named IFN-α DC） or IL-4 50 ng/ml （named IL-4 DCs） and then CD40L and LPS were added to induce DC maturation. The morphologic features of IFN-α DCs and IL-4 DCs were observed by Giemsa staining. The expressions of CDla, CD80, CD83, CD86 and HLA- DR on the surface of IFN-α DCs and IL-4 DCs were assayed by flow cytometry. The abilities of IFN-α DCs and IL-4 DCs to induce the proliferation of allogenic T cells were determined by mixed lymphocyte reaction （MLR）. Results： Both IFN-（x DCs and IL-4 DCs displayed typical DC features in morphology. The expressions of CDla, CD80, CD83, CD86 and HLA-DR in IFN-α DCs and IL-4 DCs were achieved at high levels at 3 d and 5 d after induced. Mature IFN-α DCs expressed a higher value of CD83 （ [ 78.25 ± 15.36 ] % vs [ 50.14 ± 10.24 ] %, P 〈 0.05 ） and CD86 （ [ 84.84±10. 12 ] % vs [ 62.93 ± 15.12 ] %, P 〈 0.05 ） than mature IL-4 DCs. Mature IFN-α DCs was stronger than immature IFN- α DCs on the ability to induce proliferation of allogenic T cells （P 〈 O. 05 ）. At the ratios of DCs： T cell being 1 ： 40 and 1：20, mature IFN-α DCs had a stronger ability to induce proliferation of allogeneic T cells than did mature IL-4 DCs （[39.43 ±9.211% vs [27.34 ＋10.63]%, P〈0.05; [60.31 ±7.86]% vs [48.63 ±6.25]%, P〈0.05）. Conclu- sion ： IFN-ot combined with GM-CSF can induce the differentiation of DCs from PBMCs of gastric cancer patients, which have a shorter culture period and stronger ability to induce the proliferation of allogenic T cells than traditional DCs in- duced by IL-4 and GM-CSF. It may result from the up-regulation of CD83 and CD86 expressions on IFN-α DCs.