Published online before print April 24, 2003, doi: 10.1161/01.RES.0000072971.88704.CB
© 2003 American Heart Association, Inc.
Molecular Medicine |
Tumor Necrosis Factor-
Induces Early-Onset Endothelial Adhesivity by Protein Kinase C
–Dependent Activation of Intercellular Adhesion Molecule-1
From the Department of Pharmacology, University of Illinois College of Medicine, Chicago, Ill. Current affiliation of A.R. is Department of Pediatrics, University of Rochester School of Medicine, Rochester, NY.
Correspondence to Asrar B. Malik, Distinguished Professor and Head, Department of Pharmacology, University of Illinois, College of Medicine, 835 S Wolcott Ave, Chicago, IL 60612. E-mail abmalik{at}uic.edu
We tested the hypothesis that TNF-
induces early-onset endothelial adhesivity toward PMN by activating the constitutive endothelial cell surface ICAM-1, the ß2-integrin (CD11/CD18) counter-receptor. Stimulation of human pulmonary artery endothelial cells with TNF- resulted in phosphorylation of ICAM-1 within 1 minute, a response that was sustained up to 15 minutes after TNF- challenge. We observed that TNF- induced 10-fold increase in PMN adhesion to endothelial cells in an ICAM-1–dependent manner and that this response paralleled the rapid time course of ICAM-1 phosphorylation. We also observed that the early-onset TNF-–induced endothelial adhesivity was protein synthesis–independent and associated with cell surface ICAM-1 clustering. Pretreatment of cells with the pan-PKC inhibitor, chelerythrine, prevented the activation of endothelial adhesivity. As PKC
, an atypical PKC isoform abundantly expressed in endothelial cells, is implicated in signaling TNF-–induced ICAM-1 gene transcription, we determined the possibility that PKC was involved in mediating endothelial adhesivity through ICAM-1 expression. We observed that TNF- stimulation of endothelial cells induced PKC activation and its association with ICAM-1. Inhibition of PKC by pharmacological and genetic approaches prevented the TNF-–induced phosphorylation and the clustering of the cell surface ICAM-1 as well as activation of endothelial adhesivity. Thus, TNF- induces early-onset, protein synthesis–independent expression of endothelial adhesivity by PKC-dependent phosphorylation of cell surface ICAM-1 that precedes the de novo ICAM-1 synthesis. The rapid ICAM-1 expression represents a novel mechanism for promoting the stable adhesion of PMN to endothelial cells that is needed to facilitate the early-onset transendothelial migration of PMN.
Key Words: tumor necrosis factor- • intercellular adhesion molecule-1 • endothelium • polymorphonuclear leukocyte adhesion • protein kinase C
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