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Originally published In Press as doi:10.1074/jbc.M204003200 on April 26, 2002

J. Biol. Chem., Vol. 277, Issue 28, 25032-25039, July 12, 2002
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Induction of Tissue Factor Expression in Human Endothelial Cells by CD40 Ligand Is Mediated via Activator Protein 1, Nuclear Factor kappa B, and Egr-1*

Udo BavendiekDagger , Peter LibbyDagger , Meagan KilbrideDagger , Rebecca ReynoldsDagger , Nigel Mackman§, and Uwe SchönbeckDagger

From Dagger  The Leducq Center for Cardiovascular Research, Cardiovascular Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115 and the § Department of Immunology and Vascular Biology, The Scripps Research Institute, La Jolla, California 92037

Induction of tissue factor expression in endothelial cells via ligation of CD40 probably figures prominently in the pathogenesis of prevalent inflammatory diseases, including atherosclerosis. However, the molecular mechanisms of tissue factor gene expression triggered by CD40 ligand (CD40L) in this cell type remain unknown. We demonstrate here that the tissue factor promoter region -278 bp to +121 bp contains the CD40L-responsive elements, consisting of activator protein 1 (AP-1)±, nuclear factor (NF) kappa B-, and Egr-1-binding sites. Mutations of either the AP-1- or NF-kappa B-binding sites markedly reduced the CD40L-dependent promoter activation. The AP-1 and NF-kappa B sites displayed constitutive and CD40L-enhanceable DNA binding activity, respectively. Of note, mutation of the Egr-1-binding sites, previously not associated with CD40 signaling, impaired activation of the tissue factor promoter. Accordingly, CD40L strongly induced Egr-1 protein expression and DNA binding activity to all three bindings sites. In contrast to CD40L, other established inducers of tissue factor in endothelial cells, interleukin-1beta or tumor necrosis factor alpha , did not increase the expression of Egr-1. In conclusion, induction of tissue factor gene expression in human endothelial cells by CD40L involves AP-1 and NF-kappa B as well as Egr-1, a pathway previously not implicated in CD40 signaling and distinct from that employed by certain other proinflammatory cytokines.

* This work was supported by National Institutes of Health (NHLBI) Grants HL-34636 and HL-56985 (to P. L.), by a grant from the Fondation Leducq, and by Deutsche Forschungsgemeinschaft Grant BA 1997/1-1 (to U. B.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: The Leducq Center for Cardiovascular Research, Cardiovascular Division, Dept. of Medicine, Brigham and Women's Hospital, Harvard Medical School, 221 Longwood Ave., EBRC 309, Boston, MA 02115. Tel.: 617-278-0455; Fax: 617-732-6821; E-mail: uschoenbeck@rics.bwh.harvard.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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