Published online before print April 24, 2003, doi: 10.1161/01.RES.0000072971.88704.CB
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Submitted on November 19, 2002
Revised on April 10, 2003
Accepted on April 10, 2003
Tumor Necrosis Factor-
Induces Early-Onset Endothelial Adhesivity by Protein Kinase C
-Dependent Activation of Intercellular Adhesion Molecule-1
Kamran Javaid ;
From the Department of Pharmacology (K.J., K.N.A., R.S.F., R.D.M., A.B.M.), University of Illinois College of Medicine, Chicago, Ill; and the Department of Pediatrics (A.R.), University of Rochester School of Medicine, Rochester, NY.
* To whom correspondence should be addressed. 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 
2-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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