Effect of RelB nuclear translocation induced by anti-CD40 antibody on phenotype and function of bone marrow-derived dendritic cells
Objective:To investigate the effect of anti-CD40 antibody on the phenotype and function of bone marrow dendritic cells(BMDCs)in mice and its possible mechanism.Methods:BMDCs were prepared from the tibia and femur of wild-type female healthy BALB/c mice and stimulated with dif-ferent concentrations of anti-CD40 antibodies(0,10 or 20 μg/mL)for 24 hours.Flow cytometry was used to detect the expression of BMDCs surface molecules MHC-Ⅱ,CD40,CD80 and CD86.The levels of IL-6,IL-12p70 and TNF-α in the culture supernatant of BMDCs were detected by ELISA,and the expression levels of RelA and RelB mRNA in BMDCs were detected by real-time fluores-cence quantitative PCR.In some experiments,RelB nuclear translocation inhibitor SN52(40 μg/mL)was added 30 minutes before the stimulation of anti-CD40 antibody(20 μg/mL).After 24 hours,the expression of BMDCs surface molecules and the level of cytokines in the culture supernatant were de-tected.Results:The expression levels of BMDCs surface molecules MHC-Ⅱ,CD40,CD80 and CD86,proinflammatory cytokines IL-6,IL-12p70 and TNF-α and RelB mRNA in high anti-CD40 antibody group(20 μg/mL)and low anti-CD40 antibody group(10 μg/mL)were significantly higher than those in control group(0 μg/mL)(P<0.05 or P<0.01),and the effect was dose-dependent.There was no significant difference in RelA mRNA expression among the three groups(P>0.05).In addition,the levels of MHC-Ⅱ,CD40,CD80 and CD86 on the surface of BMDCs and the levels of IL-6,IL-12p70 and TNF-α in the culture supernatant in the anti-CD40 antibody plus SN52 group were significantly lower than those in the anti-CD40 antibody group(P<0.01),and there was no sig-nificant difference between the anti-CD40 antibody plus SN52 group and the control group(P>0.05).Conclusion:Anti-CD40 antibody may promote BMDCs phenotypic and functional maturation by acti-vating RelB nuclear translocation,and the effect is dose-dependent,which may provide a new idea for disease immunotherapy.
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