CpG联合MUC1增强人外周血树突状细胞功能的实验研究

摘要

Objective:To observe the effect of CpG combined with mucin 1 (MUC1 )on the proliferation of den-dritic cells (DC)stimulated T cells.Methods:Lymphocyte separation liquid was used to isolate from human periph-eral blood mononuclear cells and cultured in vitro cell cytokines GM-CSF,IL-4 and TNF -αinduced 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+CpG,C:DC+MUC1 ,D:DC+CpG+MUC1 ,E:CpG+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. CpG+DC group and MUC1 +DC group,respectively,compared with pure DC group,T cell proliferation was signifi-cantly enhanced (P<0.01 ).CpG+MUC1 +DC was higher than that with MUC1 or CpG+DC group,differences were significant (P<0.01),the simple addition of CpG and MUC1 (DC-free group)without significantly stimula-ting the proliferation of T cells.Conclusion:CpG combined with MUC1 can significantly improve the function of DC in promoting the proliferation of T cells.%目的：观察CpG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＋CpG；C：DC＋MUC1；D：DC＋CpG＋MUC1；E：CpG＋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，CpG＋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＋CpG＋MUC1组又明显高于单用MUC1或CpG＋DC组，差别显著（P＜0．01），单纯加CpG和MUC1（无DC组）则基本无明显刺激T细胞增殖作用。结论：CpG联合MUC1能明显提高DC促T细胞增殖的功能。