Syndecan-4-dependent signaling in the inhibition of endotoxin-induced endothelial adherence of neutrophils by antithrombin

Nicole C. Kaneider; Ellen Förster; Birgit Mosheimer; Daniel H. Sturn; Christian J. Wiedermann

doi:10.1160/TH03-03-0184

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 90(06): 1150-1157
DOI: 10.1160/TH03-03-0184

Wound Healing and Inflammation/Infection

Schattauer GmbH

Syndecan-4-dependent signaling in the inhibition of endotoxin-induced endothelial adherence of neutrophils by antithrombin

Nicole C. Kaneider

¹Division of General Internal Medicine, Department of Internal Medicine, University of Innsbruck, Innsbruck, Austria

,

Ellen Förster

¹Division of General Internal Medicine, Department of Internal Medicine, University of Innsbruck, Innsbruck, Austria

,

Birgit Mosheimer

¹Division of General Internal Medicine, Department of Internal Medicine, University of Innsbruck, Innsbruck, Austria

,

Daniel H. Sturn

¹Division of General Internal Medicine, Department of Internal Medicine, University of Innsbruck, Innsbruck, Austria

,

Christian J. Wiedermann

¹Division of General Internal Medicine, Department of Internal Medicine, University of Innsbruck, Innsbruck, Austria

› Author Affiliations

Abstract

Summary

Circulating endotoxin is elevated in sepsis and plays a role in endothelial dysfunction whereas antithrombin is decreased by virtue of its consumption during complex formation with clotting factors and by proteolytic degradation by granulocyte elastase. Dysfunction of endothelium results in enhanced leukocyte rolling and diapedesis into tissues leading to edema formation and injury. Antithrombin exerts beneficial effects on endothelial function in sepsis. A direct anti-inflammatory action of anti-thrombin in inflammatory cells is exerted via heparan sulfate proteoglycans. In this study, we investigated whether antithrom-bin affects endotoxin-induced adhesion of neutrophils to human endothelial cells in vitro and whether glycosaminoglycans are involved in its signaling. Adhesion of human neutrophils to monolayers of umbilical vein endothelial cells was tested under static conditions. Endothelial cells were pretreated with endotoxin, interleukin-1, heparinase-I, chondroitinase-ABC or anti-syndecan-4-antibody. Endotoxin and interleukin-1 increased neutrophil adherence to human umbilical vein endothelial cells which was inhibited by antithrombin. Concomitant incubation with pentasaccharide abolished this effect of antithrombin. Treatment of endothelial cells with heparinase or chondroitinase led to higher adhesion and prevented effects of antithrom-bin. With antibodies to syndecan-4, enhanced adhesion of neutrophils was observed. As studied by Western blotting, endo-toxin-induced signaling was diminished by antithrombin and the effect was reversible by chondroitinase or heparinase. From our results, we can conclude that endotoxin-induced adhesion of leukocytes to endothelium can be reversed by ligation of syndecan-4 with antithrombin´s heparin-binding site and interferences with stress response signaling events in endothelium.

Keywords

Syndecan-4 - neutrophils - antithrombin

Full Text

References

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