Role of NF-κB pathway in CD40-CD154 interaction-induced human aortic endothelial cell activation

Abstract

Objective To determine human endothelial cells(VEC) activation induced by CD40-CD154 interaction and the role of NF-κB pathway in CD40-CD154 interaction. Methods VEC were isolated from aorta,and subcultures were stimulated by recombinant human TNF-α(rhTNF-α)or IFN-γ(rhIFN-γ). The expression of CD40 and adhesion molecules was detected by using flow cytometry. D1.1 cells expressing CD154 were co-incubated with VEC with or without CD154 specific monoclonal antibody, and the effects of CD40-CD154 interaction in induceding VEC activation were determined. In an additional study,BAY 11-7082,a NF-κB pathway inhibitor,was used to study whether CD40-CD154 interaction-induced VEC activation was NF-κB pathway-dependent. Results Resting VEC demonstrated weak expression of CD40,CD54 and CD106 but undetectable CD62E. VEC stimulated with rhTNF-α or rhINF-γ up-regulated the expression of CD40,CD62E,CD54 and CD106 respectively. VEC demonstrated up-regulation of CD62E, CD54 and CD106 after co-incubation with D1.1 cells,and VEC activation induced bv CD40-CD154 interaction was inhibited by CD154 specific monoclonal antibody. NF-κB could block the VEC activation induced bv rhTNF-α stimulation or by CD40-CD154 interaction. Conclusions The interaction between VEC-derived CD40 and CD154 expressing T cells plays a critical role in VEC activation. Inhibition of NF-κB during CD40-CD154 interaction prevents VEC activation. Key words: NF-κB; Endothelial cell; CD40 ligand; CD154 ligand; Transplantation