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目的研究两歧双歧杆菌完整肽聚糖(WPG)对脐血来源树突状细胞(DC)形态及分泌细胞因子的影响,了解双歧杆菌WPG对DC分化、成熟及免疫调节功能的作用;并为益生菌及其生物活性成分的进一步开发提供依据。方法分离正常孕妇脐血单个核细胞诱导生成未成熟树突状细胞(Dendritic cells,DCs),实验组在培养的第7天分别加入两歧双歧杆菌WPG(5μg/ml)、两歧双歧杆菌全菌(100μg/ml),阳性对照组加入脂多糖(LPS),阴性对照组仅加入培养基。倒置显微镜在培养各期形态学观察,流式细胞术检测表面标志物CD83及CD1a的表达,MLR检测DCs刺激同种异体T淋巴细胞能力,ELISA法测定DCs培养上清中IL-12p70、IL-10的分泌。结果脐血单核细胞在双歧杆菌WPG与GM-CSF、IL-4协同诱导作用下,能成为形态上具有典型树突状突起的DCs;诱导后的CB-MDDCs刺激同种异体T细胞的增殖能力及分泌IL-12p70、IL-10的水平显著高于阴性对照组(P<0.01)且细胞表面标志物CD83及CD1a的表达增加。结论(1)双歧杆菌WPG能影响CB-MDDCs的成熟状态。(2)双歧杆菌WPG对CB-MDDCs成熟程度及分泌细胞因子水平的影响强于双歧杆菌全菌,说明WPG是双歧杆菌主要免疫活性成分。
Objective To investigate the effect of WPG on the morphology and secretion of cytokines in cord blood derived dendritic cells (DCs) and to understand the role of WPG in Bifidobacterium on DC differentiation, maturation and immunomodulatory function. And provide the basis for the further development of probiotics and their bioactive components. Methods Cord blood mononuclear cells from normal pregnant women were induced to produce immature dendritic cells (DCs). On the 7th day of culture, experimental groups were given Bifidobacterium bifidum WPG (5μg / ml) Bacillus bacteria (100μg / ml), the positive control group added lipopolysaccharide (LPS), the negative control group only added to the medium. The expression of CD83 and CD1a on the surface of the DCs were detected by flow cytometry. The ability of DCs to stimulate allogeneic T lymphocytes was detected by MLR. The levels of IL-12p70 and IL- 10 secretion. Results Cord blood mononuclear cells could form DCs with typical dendritic protuberances under the synergistic induction of WPG, GM-CSF and IL-4, and CB-MDDCs stimulated allogeneic T cells Proliferation and secretion of IL-12p70, IL-10 were significantly higher than those in the negative control group (P <0.01), and the expression of CD83 and CD1a were increased. Conclusion (1) Bifidobacterium WPG can affect the maturation status of CB-MDDCs. (2) The influence of Bifidobacterium WPG on the maturation of CB-MDDCs and the secretion of cytokines was stronger than that of Bifidobacteria, indicating that WPG was the main immunogenic component of Bifidobacterium.