文章摘要
连利霞 彭霞 李莉 朱伟彦 安云霞 孔存权.过敏性哮喘小鼠树突状细胞负荷 Der p2 后改变及与T 细胞增殖分化的关系研究[J].,2014,14(35):6843-6848
过敏性哮喘小鼠树突状细胞负荷 Der p2 后改变及与T 细胞增殖分化的关系研究
Effects of Der p2 on DC and the Relationship between DCand T Cell Proliferation
  
DOI:
中文关键词: 过敏性哮喘  树突状细胞  Der p2
英文关键词: Allergic asthma  Dendritic cells  Der p2
基金项目:国家自然科学基金项目( 30671919; 309728248)
作者单位
连利霞 彭霞 李莉 朱伟彦 安云霞 孔存权 河南省人民医院(郑州大学人民医院) 上海交通大学第一人民医院 
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中文摘要:
      目 的: 探讨哮喘小鼠与正常小鼠骨髓源 性树突状细胞( DC)负 荷 Der p2 抗原后表达表面分子(CD11c、CD86)和细胞因子 (IL-10、IL-1 2p70)的差异及其对 Th1 和 Th2 型细胞因子平衡的影响, 进一步研究过敏性哮喘发生中 DC 的可能作用 。 方法: 分别从 哮喘组和对照组提取骨髓培养 DC, 第五天负 荷 Der p2, 24 小时后吹打收集细胞, 观察 DC 形态,用 流式细胞仪检测孵育后细胞表 面 CD1 1c、 CD86 表达。 并留取负 荷 Der f2 前后培养上清, ELISA 法检测 IL-10 及 IL-12p70 含量。 同 时以 DC: 反应细胞比例为 1: 10 混合培养, 72 h 后 ELISA 法检测混合培养上清中 IL-4、IL-5、IFN-酌 的水平。 结果: ① 负 荷 Der p2 后, 哮喘组 CD86、CD11c 表达 比对照组高, 分别 为( t=11, P<0.05)( t=4.9, P<0.05), 差异有统计学意义; ② 在细胞因子分泌方面, Der p2 负 荷前, 两组 DC 均能分 泌 IL-10 与 IL-1 2p70, IL-10 水平哮喘组高( t=9.5, P<0.05), 而 IL-12p70 水平对照组高( P<0.05); 负 荷 Der p2 后 , 对照组 IL-10、 IL-12p7 分泌量比负 荷前明显增加( P<0.05),哮喘组无明显差异( P>0.05); ③ 在 DC 刺激同 种 T 细胞因子分泌方面,负 荷 Der p2 后 哮喘组 DC 刺激 T 细胞分泌 IL-4、 IL-5 分泌能力明 显增强( P<0.05), 而刺激 INF-酌 能力降低( P<0.05)。 结论: DC 在过敏性哮喘中 起着重要作用,异常 DC 通过增加 CD86、CD11c 的表达和减少 IL-10 及 IL-12 的合成, 致使 T 细胞向 Th2 细胞优势分化。
英文摘要:
      Objective:To investigate the differences of the expression of CD11 c, CD86 and the secretion of IL-1 0 and IL-1 2p70 in the dendritic cells (DCs) from allergic asthmatics and healthy donors, which loaded the Der p2 antigen, and further study the possible role of DCs in the occurrence of allergic asthma.Methods:Marrow-derived dendritic cells (DC) were extracted and cultured, and the cells were collected after the Derp2 load 24 hours at the 5 th day, the cells morphology was observed, and the CD1 1c, CD86 expression was tested with flow cytometry cell. After and before the Derp2 loaded, the culture supernatants was collected to test the IL-10 and IL-12P70 levels with ELISA, then co-culture the DC and T cells with a proportion of 1:10 to test the levels of IL-4, IL-5 and INF-酌 in culture supernatant with ELISA.Results:① After loading Der p2, compared with control group, the expression of CD86, CD11 c in asthma group was increased with statistical significance, respectively (t=11 , P <0.05) (t=4.9, P<0.05); ② In the aspect of secretion of the cytokines, both groups could secrete IL-1 0 and IL-12P70 before Der p2 loaded, and the level of IL-10 in the asthma group was higher (t=9.5, P<0.05), while the level of IL-12p70 in the control group was higher (t=6.56, P<0.05); after Der p2 load, the level of IL-10 and IL-12p70 of the control group was increased significantly (t=19.4, P< 0.05), while asthma group had no significant difference(P>0.05). ③ In the aspect of DC stimulation on secretion of T cells, after Der p2 loaded, for the asthma group, the levels of IL-4 and IL-5 secreted by T cells by DC stimulation was significantly enhanced (P<0.05), while the level of INF-r was decreased (P <0.05).Conclusion:These results identify abnormal DC can increase CD86 and CD1 1c expression and reduce the synthesis of IL-1 0 and IL-1 2 which are necessary and sufficient for the T cells differentiation into Th2 cells.
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