可溶性虫卵抗原对小鼠骨髓源性树突状细胞表型及Th17细胞分化的影响
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国家自然科学基金(30972573),国家“863”项目资助(2006AA02Z415)


Dendritic cells stimulated by the Schistosoma japonicum soluble egg antigen induce the differentiation of Th17 cell
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    摘要:

    目的:探讨日本血吸虫可溶性虫卵抗原(soluble egg antigen,SEA)致敏小鼠骨髓源性树突状细胞(dendritic cells,DCs)后对其细胞表型及Th17细胞分化的影响-方法:收集BALB/c小鼠骨髓细胞,应用粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)-白介素(interleukin,IL)-4定向诱导小鼠骨髓细胞向DCs分化;流式细胞仪检测SEA刺激培养后DCs表面分子的表达情况;流式细胞术检测DCs 分泌IL-6-IL-23的水平,ELISA法检测DCs分泌转化生长因子-β(transforming growth factor-β,TGF-β)的水平;SEA刺激DCs与CD4+T细胞共培养后检测培养上清中IL-6-TGF-β-IL-23和IL-17细胞因子水平-结果:诱导培养出可供实验用的高纯度DCs;SEA刺激DCs能表达较高水平的CD80-CD86-CD40-组织相容性复合体(major histocompatibility complex,MHC)-Ⅱ类分子;SEA刺激DCs产生IL-6和TGF-β明显增多;SEA刺激的共培养细胞上清中IL-17水平增高-结论:SEA能够促进DCs成熟,并诱导CD4+T细胞向Th17细胞分化-

    Abstract:

    Objective:To observe the functional changes of murine bone marrow derived dendritic cells (DCs) loaded with the soluble egg antigen (SEA),and its impact on the differentiation of Th17 cell. Methods:Murine bone marrow cells were induced to differentiate into DCs by GM-CSF and IL-4. Flow cytometry was used to measure IL-6 and IL-23 and ELISA was used to measure TGF-β concentrations in the supernatant of cultured DCs. CD4+T cells and DCs were co-cultured and then the concentrations of relevant cytokines in the culture supernatant were detected. Results:Highly purified DCs were induced and could be used for further experiments. DCs were stimulated with SEA to express high levels of CD40,CD80,CD86 and MHC-II. Meanwhile,DCs stimulated with SEA produced significantly higher concentrations of IL-6 and TGF-β than the blank control group. Co-cultured cells stimulated with SEA yielded significantly higher concentrations of IL-17 than the blank control group. Conclusion:Stimulation by SEA generates mature DCs,and can induce the differentiation of CD4+T cells towards Th17 cells.

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薛 冰,徐 璐,周 亮,仇镇宁,唐 奇,李玉华,王祝鸣,朱 进,冯振卿,管晓虹.可溶性虫卵抗原对小鼠骨髓源性树突状细胞表型及Th17细胞分化的影响[J].南京医科大学学报(自然科学版),2012,(5):616-620

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