Department of Pediatrics, Chang Gung Memorial Hospital, Keelung 204, Taiwan, China.
World J Gastroenterol. 2010 Jul 28;16(28):3529-40. doi: 10.3748/wjg.v16.i28.3529.
To investigate the immune response of peripheral blood mononuclear cells (PBMCs) and dendritic cells (DCs) that were stimulated by probiotic preparations.
PBMCs were isolated, cultured, and stimulated with Bio-Three (a mixture of Bacillus mesentericus, Clostridium butyricum and Enterococcus faecalis; 10(5), 10(6) and 10(7) CFU/mL for 24 h). Cytokine production of (1) circulating PBMCs; (2) PBMCs stimulated by probiotic preparation; (3) monocyte-derived DCs; and (4) DC and T cell co-culture was determined by enzyme-linked immunosorbent assay. Phenotypic analysis of circulating PBMCs was also investigated by flow cytometry. Blood was obtained from individuals who consumed Bio-Three (10(9) CFU/d B. mesentericus, C. butyricum and E. faecalis) for 2 wk, or those who did not take probiotics orally.
In culture supernatants, interferon-gamma (IFN-gamma) and interleukin (IL)-10 production increased, but IL-4 and tumor necrosis factor-alpha (TNF-alpha) production by PBMCs decreased after 1 and 2 wk of probiotic treatment. Flow cytometry was also performed on day 14 and detected enhanced expression of CD11b, HLA-DR, CD4, CD45RA, CD25, CD44 and CD69 in response to Bio-Three. Furthermore, IL-10 and IL-12 were upregulated in supernatants of monocyte-derived DCs, and IFN-gamma and IL-10 were enhanced in supernatants of CD4(+) T cells co-cultured with DCs.
Bio-Three appeared to stimulate the Th1 immune response, downregulate pro-inflammatory cytokines (TNF-alpha) and upregulate anti-inflammatory cytokine (IL-10). Probiotics could be effective in activation of PBMCs and DCs.
研究外周血单个核细胞(PBMC)和树突状细胞(DC)对益生菌制剂刺激产生的免疫反应。
分离、培养 PBMC,并用 Bio-Three(混合了芽孢杆菌、丁酸梭菌和粪肠球菌;10(5)、10(6)和 10(7)CFU/mL,培养 24 小时)刺激。通过酶联免疫吸附试验测定(1)循环 PBMC;(2)益生菌制剂刺激的 PBMC;(3)单核细胞来源的 DC;以及(4)DC 和 T 细胞共培养的细胞因子产生情况。通过流式细胞术还研究了循环 PBMC 的表型分析。采集服用 Bio-Three(10(9)CFU/d 芽孢杆菌、丁酸梭菌和粪肠球菌)2 周的个体或未口服益生菌的个体的血液。
在培养上清液中,IFN-γ和 IL-10 的产生增加,但 PBMC 在益生菌治疗 1 周和 2 周后 IL-4 和 TNF-α的产生减少。在第 14 天也进行了流式细胞术,检测到对 Bio-Three 的反应中 CD11b、HLA-DR、CD4、CD45RA、CD25、CD44 和 CD69 的表达增强。此外,单核细胞来源的 DC 上清液中 IL-10 和 IL-12 上调,与 DC 共培养的 CD4(+)T 细胞上清液中 IFN-γ和 IL-10 增强。
Bio-Three 似乎刺激了 Th1 免疫反应,下调了促炎细胞因子(TNF-α),上调了抗炎细胞因子(IL-10)。益生菌可能有效激活 PBMC 和 DC。