Figure 1

Evaluation of adhesion molecule expression by TEC and binding of PMN to TEC and HUVEC. (a) FACS analysis of CD40 and inter-cellular adhesion molecule-1 (ICAM-1) expression by tubular epithelial cells (TEC) incubated with healthy plasma, septic plasma or septic plasma after Amberchrom resin adsorption (Septic + CG161 M). Kolomogorov Smirnov statistical analysis was performed. (b and c) In vitro adhesion assay of freshly purified polymorphonuclear neutrophils (PMN) on (b) TEC or (c) human umbilical vein endothelial cells (HUVEC) monolayers: Amberchrom resin adsorption significantly reduced septic plasma-induced PMN-TEC and PMN-HUVEC interaction (*P < 0.05 Septic vs Healthy; ^§P < 0.05 Septic + CG161 M vs. Septic). (d) Effect of the CD40/CD154 pathway on PMN/TEC interaction: PMN adhesion was significantly increased in presence of supernatants collected from chinese hamster ovary (CHO) cells transfected with a cDNA coding for a soluble form of CD154 but not with an empty vector (*P < 0.05 CHO gp39-CD8 vs. Vehicle or CHO mock) and decreased in TEC engineered by siRNA to knock-down CD40 (^§P < 0.05 Septic + CD40 siRNA vs Septic or Septic + control siRNA). Data in b and c are expressed as average number ± standard deviation fluorescent cells in 10 different fields (×100 magnification). Analysis of variance with Newmann-Keuls multicomparison test was performed.