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

Ling-ling ZHANG,Feng ZHANG,Jin-ling SHU,Ying LI,Yu-jing WU,Xian-zheng ZHANG,Le HAN,Xiao-yu TANG,Chen WANG,Yu TAI,Qing-tong WANG,Jing-yu CHEN,Yan CHANG,Hua-xun WU,Wei WEI

(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Hefei 230032,China)

T1-44

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

Ling-ling ZHANG,Feng ZHANG,Jin-ling SHU,Ying LI,Yu-jing WU,Xian-zheng ZHANG,Le HAN,Xiao-yu TANG,Chen WANG,Yu TAI,Qing-tong WANG,Jing-yu CHEN,Yan CHANG,Hua-xun WU,Wei WEI

(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Hefei 230032,China)

OBJECTIVEThis study was to investigate the effects of CP-25 on the functions of activated human B cells through regulating BAFF and TNF-alpha signaling.METHODSB cells from peripheral blood mononuclear cells(PBMCs)of normal human were isolated using magnetic cell separation(MACS)by a positive selection.B cells(107cells·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(10-5mol·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.RESULTSCP-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.CONCLUSIONCP-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.

BAFF;TNF-alpha;signaling pathway;CP-25;etanercept;rituximab

The project supported by National Natural Science Foundation of China(81330081,81473223 and 81673444);and Anhui Province Postdoctoral Science Foundation(2016B134)

Ling-ling ZHANG,Tel:13856001266,E-mail：llzhang@ahmu.edu.cn;Wei WEI,Tel:13605510563,E-mail：wwei@ahmu.edu.cn