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作 者:张云燕[1] 李有强[2] 冉炜[1] 黄彬[1] 郭冰[1] 廖鑫[1] 陈茶[2] 屈平华[2] 张轩[2]
机构地区:[1]中山大学附属第一医院, 广州501180 [2]广东省中医院检验科, 广州510006
出 处:《中华微生物学和免疫学杂志》2013年第3期197-200,共4页Chinese Journal of Microbiology and Immunology
基 金:国家自然科学基金(81071397,81271909);广东省科技计划项目(20118031800037)
摘 要:目的探讨铜绿假单胞菌分泌的信号分子3-O-C12-HSL对脂多糖诱导的人外周血单核细胞来源树突状细胞(Mo.DCs)成熟及n细胞极化的干预作用。方法采用免疫磁珠法分选人外周血CDl4+单核细胞,经重组人粒一单核细胞集落刺激因子和重组人IL-4诱导其分化为Mo-DCs。CCK-8法检测3-0.CIrHSL对Mo.DCs活性影响;流式细胞术检测3-O-C12-HSL对Mo-DCs表型(CDlle-,CD40、CD80、HIJA.DR)的影响;ELISA检测Mo—DCs培养上清IFN-1和IL-12浓度;混合淋巴细胞反应观察3-O.C旷HSL对Th细胞增殖和细胞因子分泌的影响。结果3-O-C12-HSL终浓度200μmol/L时可影响Mo-DCs活性。与脂多糖对照组比较,终浓度50μmoL/L和100μmoL/L的3-O-C12-HSL明显下调脂多糖诱导的Mo-DCs表面分子CD40、CD80和HLA-DR分子表达水平,抑制Mo·DCs分泌IL-12,促进Mo.DCs分泌IL-10,抑制Th细胞增殖和IFN-γIL-12分泌,促进n细胞分泌IL-10。结论3-O-C12-HSL可抑制脂多糖诱导的Mo-DCs成熟,诱导Th2细胞极性分化,影响机体免疫系统对铜绿假单胞菌的清除作用。Objective To investigate the influence of 3-O-C12-HSL, which secreted by Pseudo- monas aeruginosa, on the maturation of monocyte-derived dendritic ceils from human peripheral blood and Th cell polarization. Methods Human peripheral blood CD14+ monocytes were separated by immunomagnetic beads and induced by human granulocyte monocyte colony stimulating factor and human intedeukin 4 in cul- ture. CCK-8 method was used to detect the influence of 3-O-C12-HSL on Mo-DCs activity. The immunophe- notypic expression of CD1 l c, CD40, CD80 and HLA-DR expression were assessed by FACS. IL-10 and IL- l2 levels in the supematant were measured by ELISA. Mixed culture of CIM T lymphocytes and Mo-DCs was used to observe the effects of 3-O-C12-HSL on the proliferation and eytokine secretion of Th cells. Results 3-O-C12-HSL affects the activity of Mo-DCs at the concentration of 200 μmol/L. 3-O-C12-HSL significantly down-regnlated the expression of cell surface molecules including CD80, CD40 and HLA-DR on Mo-DCs at the concentration of 50 μmol/L and 100 μmol/L. The secretion of IL-10 on Mo-DCs was in- creased whereas intedeukin-12 was decreased. The proliferation and IFN-γ, IL-12 secretion of Th cells were inhibited by 3-O-C22-HSL. However, IL-10 expression was enhanced. Conclusion 3-O-C12-HSL inhibited lipopolysaeeharide-induced maturation of Mo-DCs and drove Th2 cell polarization. Then, it might further affect the host immune system for clearance of Pseudomonas aeruginosa.
关 键 词:3-O-C12-HSL 树突状细胞 Th细胞极性分化
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