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The Journal of Immunology, 2000, 164: 3316-3322.
Copyright © 2000 by The American Association of Immunologists

A Functional Role of I{kappa}B-{epsilon} in Endothelial Cell Activation1

Martin Spiecker*, Harald Darius* and James K. Liao2,{dagger}

* Vascular Medicine and Atherosclerosis Unit, Cardiovascular Division, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115; and {dagger} Medical Clinic II, Johannes Gutenberg University, Mainz, Germany

The NF-{kappa}B inhibitor I{kappa}B-{epsilon} is a new member of the I{kappa}B protein family, but its functional role in regulating NF-{kappa}B-mediated induction of adhesion molecule expression is unknown. In vascular endothelial cells, I{kappa}B-{epsilon} associates predominantly with the NF-{kappa}B subunit Rel A and to a lesser extent with c-Rel, whereas I{kappa}B-{alpha} and I{kappa}B-ß associate with Rel A only. Following stimulation with TNF-{alpha}, pyrrolidine dithiocarbamate (PDTC), N-acetylcysteine, and dexamethasone prevented I{kappa}B kinase-induced I{kappa}B-{alpha}, but not I{kappa}B-ß or I{kappa}B-{epsilon} phosphorylation and degradation. Since the activation of NF-{kappa}B is required for the induction of adhesion molecule expression, we examined the role of I{kappa}B-{epsilon} in the transactivation of promoters from VCAM-1, ICAM-1, and E-selectin. Using reporter gene constructs of adhesion molecule promoters, PDTC inhibited VCAM-1 and E-selectin, but to a lesser extent, ICAM-1 promoter activity. Subcloning of {kappa}B cis-acting elements of VCAM-1, E-selectin, and ICAM-1 into a heterologous promoter construct revealed that PDTC inhibited VCAM-1 and E-selectin, but to a lesser extent, ICAM-1 {kappa}B promoter activity. By electrophoretic mobility shift assay, NF-{kappa}B heterodimers containing c-Rel specifically bind to the {kappa}B motif in the ICAM-1, but not VCAM-1 or E-selectin promoter. Indeed, overexpression of c-Rel induced ICAM-1 {kappa}B promoter activity to a greater extent than that of E-selectin and overexpression of I{kappa}B-{epsilon} inhibited ICAM-1 and VCAM-1 promoter activity in endothelial cells. These findings indicate that c-Rel-associated I{kappa}B-{epsilon} is involved in the induction of ICAM-1 expression.




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