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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-12p70)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CD11c、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-γ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-12p70,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-γ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。
Objective: To investigate the difference of expression of surface molecules (CD11c, CD86) and cytokines (IL-10, IL-12p70) after the Der p2 antigen loaded on bone marrow-derived dendritic cells (DCs) Th1 and Th2 cytokine balance, and further study the possible role of DC in allergic asthma. Methods: DCs were harvested from the asthma group and the control group, Der p2 was loaded on the fifth day, and collected after 24 hours. The morphological changes of DCs were observed. The expression of CD11c and CD86 on the cell surface was detected by flow cytometry. The culture supernatants were collected before and after Der f2 loading, and the levels of IL-10 and IL-12p70 were detected by ELISA. At the same time, the ratio of DC to reactive cells was 1:10, and the levels of IL-4, IL-5 and IFN-γ in mixed culture supernatants were detected by ELISA after 72 h. Results: The expression of CD86 and CD11c in Asthma group was significantly higher than that in the control group (t = 11, P <0.05) at 1 load Der p2 (t = 4.9, Before Der p2 stress, both DCs secreted IL-10 and IL-12p70, IL-10 level asthma group was higher (t = 9.5, P <0.05), while IL-12p70 level was higher in control group (P < 0.05). After Der p2 was loaded, the secretion of IL-10 and IL-12p7 in the control group was significantly increased (P <0.05) compared with that before the load, but there was no significant difference in the asthma group (P> 0.05) On the secretion side, DCs secreting IL-4 and IL-5 secreted by DC stimulated by DCs with Der p2 significantly increased (P <0.05) and INF-γ decreased (P <0.05). Conclusion: DC plays an important role in allergic asthma. Abnormal DC can differentiate T cells to Th2 cells by increasing the expression of CD86 and CD11c and decreasing the synthesis of IL-10 and IL-12.