热休克蛋白60对小鼠树突状细胞成熟及免疫功能的影响

Effect of Heat Shock Protein 60 on Murine Dendritic Cell Maturation and Immunologic Function

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)功能的影响。方法从近交系C57BL/6J小鼠骨髓中提取mDC,体外培养成熟后分别与3、10μg/ml mHSP60或PBS孵育24 h,观察其形态改变;流式细胞仪检测mDC表面标志;混合淋巴细胞反应(MLR)测定mDC刺激功能;ELISA法测定MLR上清液中细胞因子浓度。结果mHSP60处理组较对照组,mDC突起增加明显;CD80及CD86表型显著增加;淋巴细胞刺激功能明显增强;上清液细胞因子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。以上变化mHSP60高剂量处理组较低剂量处理组更明显。结论mHSP60体外可以促进mDC功能,且作用呈剂量依赖性。

Objective To investigate the effect of heat shock protein 60 （HSP60）, an important inflammatory factor in atherogenesis, on the maturation of murine dendritic cells （mDCs） in vitro, Methods Myeloid mDCs were obtained in vitro from C57BL/6J mouse marrow and incubated to mature with 3 μg/ml and 10 μg/ml of mHSP60 or PBS for 24 h. The morphological changes were observed. The phenotype was detected by using flow cytometry. The stimulating capacity was determined by allogenic mixed lymphocyte reaction （MLR）. ELISA was used to measure the level of cytokines in thesupernatant of MLR. Results Compared to control group, mDC showed more protrusions, the expression of CD86 and CD80 on mHSP-DC significantly up-regulated, the stimulating capacity of mHSP-DCs obviously enhanced, levels of IFN-γ in the supernatant increased. and the ratio of IFN-γ/IL-4 elevated in mHSP60-treated groups. The above changes were more significant in the high dose mHSP60-treated group than in the low dose mHSP60-treated group. Conclusion mHSP60 could promote the maturation of mDC dose-dependently in vitro.