重组HSP60蛋白体外对小鼠骨髓树突状细胞功能的影响

Impact of HSP60 on function of dendritic cells cultured from murine bone marrow in vitro

目的:观察重组热休克蛋白60(HSP60)对小鼠骨髓树突状细胞(DC)功能的影响,探讨其可能的作用机制。方法:培养小鼠骨髓源性DC,分为对照组及HSP60组,对照组在正常条件下培养,HSP60组加入终浓度为10μg/ml的HSP60。流式细胞术检测DC细胞表面CD80、CD86、MHCⅡ分子、CD14及TLR4的变化,ELISA法检测DC分泌TNF-α、IFN-γ和IL-12的浓度,免疫细胞化学检测DC MyD88、核因子-κB(NF-κB)的表达,混合淋巴细胞培养检测T细胞增殖能力。结果:HSP60可促进DC高表达CD80、CD86、MHCⅡ分子、CD14及TLR4,促进Th1型细胞因子TNF-α、IFN-γ及IL-12释放、MyD88高表达及NF-κB核移位,并诱导T细胞增殖。结论:HSP60可促进DC成熟,其作用机制可能与激活了TLR4信号通路有关。

Objective：To explore the impact and mechanism of HSP60 on maturation of dendritic ceils （DCs） cultured fl＇om murine bone marrow. Methods： Mouse DCs were generated from bone malTow ceils and were divided into control group and HSP60 group. DCs in control group was cultured at normal condition, and HSP60 group was added HSP60 of the final concentrations of 10 μg/ ml. The CD80,CD86, MHC II ,CD14 and TLR4 were detected by flow cytometry. ELISA was used to detect the concentration of TNF-α, IFN-＇y and IL-12 in the supernatant. Immunochemistry was used to detect the concentration of MyD88 and NF-KB. Mixed lympho- cyte reaction （MLR） was used to detect the functional properties of DCs. Results： HSP60 increased the CD80, CD86, MHC 11 , CD14 and TLR4 in the cytomembrane of DCs and TNF-ct, IFN-y, IL-12 concentration in the supernatant. HSP60 also promoted the expres- sion of MyD88 and the shift of NF-KB to karyon. HSP60 could also induce the proliferation of T cells. Conclusion： HSP60 perhaps promotes the maturation of DCs through Toll-like receptor 4 signal pathway.