白细胞介素10 通过抑制自噬调节树突状细胞功能

目的:研究白细胞介素10(Interleukin-10,IL-10)对树突状细胞(Dendritic cells,DCs)自噬及功能的影响。方法:体外培养C57BL/6 小鼠骨髓来源的DCs 分为对照组、LPS 刺激组、IL-10 干预组、IL-10 与雷帕霉素干预组、雷帕霉素干预组,分别通过流式细胞术分析DCs 表面共刺激分子CD40、CD80 的表达、DCs 摄取OVA 抗原的比例以及诱导OT2 细胞增殖比例,ELISA 检测DCs 分泌IL-6,TNF-β的水平等DCs 相关功能;蛋白免疫印迹法检测DCs 自噬蛋白LC3 的表达,比较组间差异,以探讨IL-10 对DCs 功能及自噬的影响。结果:(1)与LPS 刺激组比较,IL-10 处理组DCs 的表面共刺激分子CD40、CD80的表达下降、分泌IL-6、TNF-β水平下降、刺激T 细胞增殖的比例明显减弱、摄取OVA 抗原的能力增加,IL-10+雷帕霉素干预组的DCs 与IL-10 单独处理组相比,表面CD80 的表达明显增加(P<0.05)、分泌IL-6、TNF-β能力及刺激T 细胞增殖的能力均明显增加(P<0.000 1)。(2)DCs 的自噬相关蛋白(LC3 / LC3 比例)明显下降。结论:IL-10 可能通过抑制DCs 自噬水平调节树突状细胞功能。

Interleukin-10 regulates functions of dendritic cell through autophagy inhibition

Objective:To study the mechanism of interleukin-10(IL-10)inhibiting the function of dendritic cells(DCs). Methods: Cultured C57BL/6 mouse bone marrow-derived DCs were divided into 5 groups:control group,LPS stimulated group,IL-10 treated group, IL-10+Rapamycin treated group and Rapamycin treated group.The regulatory mechanism of IL-10 on dendritic cells were evaluated from DCs function,Flow cytometry was used to analyse the expression of DCs surface co-stimulator CD80,CD40 expression,the ability of uptaking antigen and stimulating T cell to proliferate;ELISA was used to detect the cytokines IL-6 and TNF-β.Western blot was used to analyse the autophagy related protein LC3.Compared the differences between the groups.Results: (1)Compared to LPS stimulated group,IL-10 treated group,DCs surface co-stimulator CD40,CD80 were decreased,IL-6 and TNF-βsecretion level and the ability to stimulate T cell to proliferate were decreased,the ability to capture OVA antigen was increased.Compared to IL-10 treated group,the DCs surface co-stimulator CD80 was decreased(P<0.05),IL-6 and TNF-βsecretion level and the ability to stimulate T cell to proliferate were increased(P<0.000 1) in IL-10+Rapamycin treated group.In addition,autophagy related proteins LC3 / LC3 was decreased in IL-10 treated group.Conclusion: IL-10 may regulate functions of DCs through inhibiting the autophagy of DCs.