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Objective: To explore the role and mechanism of myeloid differentiation factor88(MyD88) in HSP60 signal transduction in dendritic cells. Methods:Mouse DCs were cultured from murine bone marrow cells. The DC marker CD11c was detected by flow cytometry, then DCs were divided into control group, HSP60 groupand RNA interference group. Control group was cultured under normal condition, and HSP60 group was cultured with 10 μg/ml of HSP60. RNA interference group was first cultured with MyD88 siRNA for12 hours and then HSP60 was added into the culture mixture. All groups were cultured for 48 hours. Immunochemistry was used to detect the concentration of MyD88 and NF- κB. Western blot was used to detect the concentration of MyD88. Flow cytometry and mixed lymphocyte reaction(MLR) were used to detect the phenotype and functional properties of DCs. ELISA was used to detect the concentration of TNF-α, IFN-r and IL-12 in the supernatant. Results:The expression of CD11c in murine bone marrow DCs was 88.76%. HSP60 stimulation increased the expression of CD80, CD86, MHC-Ⅱ in DCs and TNF-α, IFN-r, IL-12 secretion in the supernatant. HSP60 stimulation also increased the level of MyD88 in the cytoplasm and promoted the shift of NF-κB to karyon and the proliferation of allogeneic T cells. MyD88 siRNA could decrease MyD88 and inhibit these effects induced by HSP60. Conclusion:HSP60 activates DCs through MyD88-dependent pathway. MyD88 plays a critical role in HSP60 signal transduction. Inhibition of MyD88 may be a novel way for treating disease correlated with HSP60.
Objective: To explore the role and mechanism of myeloid differentiation factor 88 (MyD88) in HSP60 signal transduction in dendritic cells. Methods: Mouse DCs were cultured from murine bone marrow cells. The DC marker CD11c was detected by flow cytometry, then DCs were divided into control group, HSP60 group and RNA interference group. Control group was cultured under normal condition, and HSP60 group was cultured with 10 μg / ml of HSP60. RNA interference group was first cultured with MyD88 siRNA for 12 hours and then HSP60 was added into the culture mixture . All groups were cultured for 48 hours. Immunochemistry was used to detect the concentration of MyD88 and NF- κB. Western blot was used to detect the concentration of MyD88. Flow cytometry and mixed lymphocyte reaction (MLR) were used to detect the phenotype and functional properties of DCs. ELISA was used to detect the concentration of TNF-α, IFN-r and IL-12 in the supernatant. Results: The expression of CD11c in murine bone marrow DCs was 88.76%. HSP60 stimulation increased the expression of CD80, CD86, MHC-II in DCs and TNF-α, IFN-r, IL-12 secretion in the supernatant. HSP60 stimulation also increased the level of MyD88 in the cytoplasm and promoted the Shift of NF-κB to karyon and the proliferation of allogeneic T cells. MyD88 siRNA could decrease MyD88 and inhibit these effects induced by HSP60. Conclusion: HSP60 activates DCs through MyD88-dependent pathway. MyD88 plays a critical role in HSP60 signal transduction. Inhibition of MyD88 may be a novel way for treating disease correlated with HSP60.