Inhibition effect of MyD88 siRNA to the activation of dendritic cells from murine bone marrow

Objective:To observe the impact of myeloid differentiaion factor 88(MyD88) siRNA on the biological activities of dendritic cells(DCs) cultured from murine bone marrow under stimulation by Lipopolysaccharides(LPS), and provide novel ideas and basis of DC’s clinical applications.Methods:Mouse DCs were generated from bone marrow cells and were divided into control group, LPS group and RNA interference group. Control group was added nothing, and LPS group was added LPS at the final concentration of 1 μg/ml. MyD88 siRNA was added in RNA interference group, and 12 hours later LPS at the final concentration of 10 μg/ml was added. All groups were cultured for 3 days thereafter. 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) was used to detect the phenotypes and functional properties of DCs. ELISA was used to detect the concentration of TNF-α, IFN-γ and IL-12 in the supernatant.Results:LPS stimulation increased the CD80, CD86, MHC-Ⅱ on the membrane of DCs and TNF-α, IFN-γ, IL-12 concentration in the supernatant. LPS stimulation also increased the MyD88 in the cytoplasm and promoted translocation of NF-κB to karyon. MyD88 siRNA can inhibit all these responess.Conclusion:MyD88 siRNA can suppress maturation of DCs and promote the tolerance-induction effect of DCs.