热休克蛋白60对小鼠树突状细胞功能影响体外研究

Effect of heat shock protein 60 on murine dendritic cell function in vitro

目的探讨动脉粥样硬化中重要炎性物质——热休克蛋白60(HSP60)体外对小鼠树突状细胞(mDC)功能影响。方法小鼠骨髓提取DC,体外培养成熟后与两种浓度mHSP60孵育,动态观察DC突起改变;流式细胞仪检测孵育前后mDC表面标志改变;MLR测定孵育前后mDC刺激功能变化;ELISA法测定MLR上清液中细胞因子浓度。结果孵育后,mDC突起增加明显;CD11c+、CD80及CD86表型显著增加;淋巴细胞刺激功能明显增强;分泌细胞因子IL-12、IFN-γ增加(P<0.01)而IL-4增加不明显(P>0.05),IFN-γ/IL-4比值升高。结论mHSP60体外可以促进mDC功能,作用呈剂量依赖性。

Objective ：To investigate the effect of heat shock protein 60 （HSP60）, significant inflammatory factor in atherosclerosis, on the maturation of murine dendritic cells （mDCs） in vitro. Methods ： Myeloid mDCs were extracted in vitro from mouse marrow and incubated to mature with two distinct concentrations of mHSP60. The morphological changes were observed;the variation of phenotype was detected with flow cytometric analysis; the stimulating capacity was determined in allogenic mixed lymphocyte reaction （MLR） ; ELISA was used to analyze the level of cytokines secreted by mDCs and stimulated lymphocyte in the medium of MLR. Restilts： Dendrites of mHSP60-DCs raised obviously. The expression of co-stimulating factor CD11c^＋ , CD86, CD80 was up-regulated. The stimulating capacity of mHSP-DCs in MLR was higher. T lymphocytes co-cuhured with mHSP-DCs in MLR secreted higher levels of pro-inflammation cytokines（ IFN-γ, IL-12） but there were no significant difference of IL-4, the stimulatory action was dose dependent and was evident at higher concentrations. The ratio of IFN-γ/IL-4 was increased. Conclusion ： mHSP60 progressed the maturation of mDC dose-dependently in vitro.