CpG联合MUC1增强人外周血树突状细胞功能的实验研究

Experimental study on the function of CpG combined with MUC1 in enhancing dendritic cell function in human peripheral blood

目的:观察Cp G联合黏蛋白1(MUC1)应用对树突状细胞(DC)刺激T细胞增殖作用的影响。方法:应用淋巴细胞分离液分离人外周血单核细胞,体外培养应用GM-CSF、IL-4、TNF-α等细胞因子诱导DC,观察细胞形态,并通过流式细胞术检测细胞标志物CD80、CD86。收获DC按如下分组:A:DC;B:DC+Cp G;C:DC+MUC1;D:DC+Cp G+MUC1;E:Cp G+MUC1,分别与T淋巴细胞混合培养,通过MTT法检测T细胞增殖情况。结果:DC表型检测结果为:CD80^+细胞占70.4%,CD86^+细胞占72.0%,呈现成熟DC表型。MUC1与DC联合疫苗与淋巴细胞混合培养显示:DC具有刺激T细胞增殖的作用,Cp G+DC组和MUC1+DC组,分别较单纯DC组对T细胞的刺激增殖作用明显增强(P<0.01),DC+Cp G+MUC1组又明显高于单用MUC1或Cp G+DC组,差别显著(P<0.01),单纯加Cp G和MUC1(无DC组)则基本无明显刺激T细胞增殖作用。结论:Cp G联合MUC1能明显提高DC促T细胞增殖的功能。

Objective： To observe the effect of Cp G combined with mucin 1（ MUC1） on the proliferation of dendritic cells（ DC） stimulated T cells. Methods： Lymphocyte separation liquid was used to isolate from human peripheral blood mononuclear cells and cultured in vitro cell cytokines GM- CSF,IL- 4 and TNF- α induced DC. Cell morphology was observed by flow cytometry cell markers CD80 and CD86. Harvest DC according to the following groups was divided into A： DC,B： DC ＋ Cp G,C： DC ＋ MUC1,D： DC ＋ Cp G ＋ MUC1,E： Cp G ＋ MUC1,respectively. T lymphocyte was in mixed culture,the proliferation of T cells by MTT assay. Results： DC phenotype detection results：CD80＋cells accounted for 70. 4 %,CD86＋cells accounted for 72. 0 %,showing a mature DC phenotype. MUC1 and DC combined vaccine and mixed lymphocyte culture showed that DC had the role of T cell proliferation stimulation.Cp G ＋ DC group and MUC1 ＋ DC group,respectively,compared with pure DC group,T cell proliferation was significantly enhanced（ P〈0. 01）. Cp G ＋ MUC1 ＋ DC was higher than that with MUC1 or Cp G ＋ DC group,differences were significant（ P〈0. 01）,the simple addition of Cp G and MUC1（ DC- free group） without significantly stimulating the proliferation of T cells. Conclusion： Cp G combined with MUC1 can significantly improve the function of DC in promoting the proliferation of T cells.