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目的:探讨IL-17A对小鼠骨髓细胞衍生树突状细胞分化和成熟的影响。方法:分离小鼠骨髓细胞,加入含GM-CSF(20 ng/ml)RPMI1640完全培基培养8 d,诱导小鼠骨髓单个核细胞向DC分化,加入LPS(1μg/ml)继续培养36 h,进一步诱导DC成熟,同时在骨髓细胞衍生诱导DC分化及成熟的不同阶段加入不同浓度的rm IL-17A(10、100 ng/ml),采用流式细胞术检测DC表面共刺激分子的表达,ELISA方法检测DC培养上清中IL-12p40和IL-10水平。结果:rm IL-17A可促进GMCSF诱导骨髓细胞衍生DC表面共刺激分子CD40、CD80、CD86和MHCⅡ的表达,且具有剂量依赖性,其中以高浓度rm IL-17A刺激组的CD40及MHCⅡ表达增加最显著;在LPS诱导DC成熟阶段加入rm IL-17A,骨髓细胞衍生DC共刺激分子CD40、CD80、CD86和MHCⅡ的表达均明显增加,并且随着rm IL-17A浓度的增加,CD86和MHCⅡ的表达水平也随之增高;同时与未加rm IL-17A的对照组相比,低浓度rm IL-17A组LPS刺激骨髓细胞衍生DC分泌IL-12p40和IL-10水平均显著增加(P<0.001),高浓度rm IL-17A组IL-12p40水平显著增高(P<0.001),但IL-10水平没有变化。结论:IL-17A可促进GM-CSF诱导的骨髓细胞衍生DC前体细胞表型发展,并能协同LPS诱导骨髓衍生DC的分化和成熟。
Objective: To investigate the effect of IL-17A on the differentiation and maturation of mouse bone marrow-derived dendritic cells. Methods: Bone marrow mononuclear cells were induced to differentiate into DCs by RPMI1640 supplemented with GM-CSF (20 ng / ml) for 8 days. Cells were cultured in LPS (1 μg / ml) for 36 hours. Furthermore, DCs were induced to mature. At the same time, different concentrations of rm IL-17A (10,100 ng / ml) were added in different stages of differentiation and maturation of DCs induced by bone marrow cells. Flow cytometry was used to detect the expression of costimulatory molecules on DC surface. Methods The levels of IL-12p40 and IL-10 in DC culture supernatants were detected. Results: rm IL-17A promoted the expression of CD40, CD80, CD86 and MHCⅡ in bone marrow-derived DCs induced by GMCSF in a dose-dependent manner. The expression of CD40 and MHCⅡ was increased in rm IL-17A stimulated group The expression of CD40, CD80, CD86 and MHCⅡ in bone marrow-derived DCs increased significantly, and with the increase of rm IL-17A, the expression of CD86 and MHCⅡ The level of IL-12p40 and IL-10 secreted by LPS stimulated by LPS stimulated by low concentration rm IL-17A group increased significantly (P <0.001 compared with the control group without rm IL-17A ), IL-12p40 was significantly increased in high concentration rm IL-17A group (P <0.001), but IL-10 level did not change. Conclusion: IL-17A can promote the phenotype of bone marrow-derived DC precursors induced by GM-CSF and can induce the differentiation and maturation of bone marrow-derived DCs in coordination with LPS.