IL-17A协同GM-CSF和LPS促进骨髓细胞衍生树突状细胞的分化和成熟研究

IL-17A promotes differentiation and maturation of bone marrow-derived dendritic cells by cooperating with GM-CSF and LPS

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-17 A on the differentiation and maturation of murine bone marrow-derived dendritic cells（ BMDCs）. Methods： Murine bone marrow cells were isolated and cultured in RPMI1640 complete medium in the presence of GM-CSF（ 20 ng / ml） for 8 days to induce differentiation of murine bone marrow cells to DC progenitors. Then these cells were treated with LPS（ 1 μg / ml） for 36 h which polarized immature DCs into mature DCs. Different concentrations of rm IL-17A（ 10 or 100 ng / ml） was added to the culture medium at different stages of BMDC differentiation and maturation. Co-stimulatory molecules expression on BMDC were analyzed by flow cytometry,and the culture supernatants were analyzed for IL-12p40 and IL-10 level by ELISA. Results： rm IL-17 could promote co-stimulatory molecules（ CD40,CD80,CD86 and MHCⅡ） expression on BMDCs in a doesdependent manner,especially,the expression of CD40 and MHCⅡ had a significant increase in high concentration of rm IL-17 A group;rm IL-17 A was added while LPS induced maturation of BMDCs. CD40,CD80,CD86 and MHCⅡ expression on BMDC increased sharply in LPS plus rm IL-17 A stimulation group,besides,CD86,MHCⅡ showed a higher level expression on BMDC with the increase of concentration of rm IL-17 A. Furthermore,secretion of IL-12p40 and IL-10 increased significantly in the group of DCs treated with LPS plus low concentration of rm IL-17 compared with the group without rm IL-17（ P 〈0. 001）. However,high concentration of rm IL-17 A group showed significantly higher levels of IL-12p40（ P〈0. 001）,but there was no difference in IL-10. Conclusion： IL-17 A promotes the phenotypic development of BMDC progenitors propagated in GM-CSF and cooperate with LPS to induce BMDC differentiation and maturation.