Role of nitric oxide in poly(I-C)-induced endothelial cell expression of leukocyte adhesion molecules.

Polyinosinic-polycytidylic acid [poly(I-C)] is a synthetic double-stranded RNA (dsRNA) that simulates a viral-infected state in cells. It has been shown that viral infection, as well as poly(I-C), stimulates leukocyte adhesion to endothelial cell (EC) monolayers and that this is mediated through the surface expression of the adhesion molecules E-selectin, vascular cell adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule 1. We have tested the involvement of nitric oxide (NO) in poly(I-C)-induced monocytic cell adhesion to human vascular EC. Using primary cultured EC for these studies, we confirmed the results from previous reports that these cells have higher basal levels of NO production than passaged cells. Poly(I-C)-induced monocytic cell adhesion to primary EC was concentration-dependently inhibited by 40-74% by the nitric oxide synthase (NOS) inhibitor NG-methyl-l-arginine (l-NMA), as well as three other NOS inhibitors, without significantly affecting interleukin-1β-induced adhesion.l-NMA inhibited poly(I-C)-induced surface expression of E-selectin and VCAM-1 by 25 and 45%, respectively, and mRNA levels of E-selectin and VCAM-1 by 62 and 74%, respectively. Primary EC transiently transfected with a plasmid containing an E-selectin promoter-driven luciferase reporter gene showed that l-NMA treatment reduced poly(I-C)-induced E-selectin promoter activity to basal levels. Electrophoretic mobility shift analysis indicated that poly(I-C)-induced nuclear factor-κB (NF-κB) binding to a radiolabeled oligonucleotide corresponding to the consensus NF-κB binding domain of the E-selectin promoter was decreased byl-NMA pretreatment. Hence, NO appears to augment E-selectin gene expression in response to poly(I-C) at the transcriptional level in vascular EC. Collectively, these data support the hypothesis that NO augments poly(I-C)-induced EC activation. These data suggest a novel role for NO as a response mediator in dsRNA-induced leukocyte adhesion to EC.