CP-25 inhibits the functions of activated human B cells through regulating BAFF-TRAF2-NF-κB and TNF-alpha-TRAF2-NF-κB signaling

OBJECTIVE This study was to investigate the effects of CP-25 on the functions of activated human B cells through regulating BAFF and TNF-alpha signaling.METHODS B cells from peripheral blood mononuclear cells(PBMCs) of normal human were isolated using magnetic cell separation(MACS) by a positive selection.B cells(107 cells·mL^(-1)) were stimulated by BAFF(100 ng·mL^(-1))or TNF-alpha(100 ng·mL^(-1)) for two hours,and then were treated with CP-25(10-5 mol·L^(-1)) or Rituximab(5 μg·mL^(-1)) or Etanercept(10 μg·mL^(-1)).B cell proliferation was detected by CCK-8.B cell subsets and BAFF receptors(BAFFR,BCMA and TACI) were analyzed by flow cytometry.The expression of TNFR1 and TNFR2 on B cells was analyzed by flow cytometry.The expression of MKK3,MKK6,P-p38,P-p65,TRAF2 and p100/52 was analyzed by Western blotting.RESULTS CP-25 inhibited B cells proliferation stimulated by BAFF or TNF-alpha.CP-25,Rituximab and Etanercept reduced the percentage and numbers of CD19^+B cells,CD19^+CD20^+B cells,CD19^+CD27^+B cells and CD19^+CD20^+CD27^+B cells induced by BAFF or TNF-alpha.CP-25 down-regulated the high expression of BAFFR,BCMA and TACI stimulated by BAFF or TNF-alpha.CP-25,Rituximab and Etanercept down-regulated significantly the expression of TNFR1 and TNFR2 on B cell stimulated by BAFF or TNF-alpha.CP-25,Rituximab and Etanercept down-regulated the expression of MKK3,P-p38,P-p65,TRAF2 and p52 in B cells stimulated by BAFF and the expression of TRAF2 and P-p65 in B cells stimulated by TNF-alpha.CONCLUSION CP-25 regulated moderately activated B cells function by by regulating the classical and alternative NF-κB signaling pathway mediated by BAFF and TNF-alpha-TRAF2-NF-κB signaling pathway.This study suggests that CP-25 may be a promising anti-inflammatory immune and soft regulation drug.

Ginsenoside metabolite compound K alleviate collegen-induced arthritis through impairing dendritic cells function

OBJECTIVE Ginsenoside metabolite compound K(CK)is a degradation product of ginsenoside in the intestine by bacteria.The anti-inflammatory and immunomodulatory activities of CK have been reported.This study investigated whether CK exerted its immunoregulatory effect through modulation of dendritic cells(DCs)function.METHODS In vivo,severity of collegen-induced arthritis(CIA),T cells and DCs subsets,phenotype of DC were assayed by flow cytometry,CCL19 and CCL21 level in lymph nodes assayed by ELISA.In vitro,bone marrow-derived DCs from normal mice were matured with lipopolysaccharide and treated with CK for 48 h.In vivo,bone marrow-derived DCs were generated from CIA mice before and 2 weeks into CK treatment.DCs were analyzed for migration,phenotype and T-cell stimulatory capacity.RESULTS CK alleviated the severity of CIA,decreased pD Cs and mo-DCs,increased na?ve T cells in CIA mice lymph nodes,and suppressed CCL21 expression in lymph nodes.CK suppressed DCs migration induced by CCL21 and T cells-stimulatory capability of DC,down-regulated LPS-induced expression of CD80,CD86,MHCII and CCR7 on DCs.CONCLUSION This study elucidated the novel immunomodulatory property of CK via impairing function of DCs in priming T cells activation.These results provide an interesting novel insight into the potential mechanism by which CK contribute to the restoration of immunoregulation in autoimmune conditions.