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

Authors

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

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

References
1 Gotte M, Joussen AM, Klein C. et al. Role of syndecan-1 in leukocyte-endothelial interactions in the ocular vasculature. Invest Ophthalmol Vis Sci 2002; 43: 1135-41.
2 Hoogewerf AJ. et al. Glycosaminoglycans mediate cell surface oligomerization of chemo-kines. Biochemistry 1997; 36: 13570-8.
3 Webb LM, Ehrengruber MU, Clark-Lewis I. et al. Binding to heparan sulfate or heparin enhances neutrophil responses to interleukin 8. Proc Natl Acad Sci USA 1993; 90: 7158-62.
4 Peter K. et al. Heparin inhibits ligand binding to the leukocyte integrin Mac-1 (CD11b/ CD18). Circulation 1999; 100: 1533-9.
5 Diamond MS. et al. Heparin is an adhesive ligand for the leukocyte integrin Mac-1 (CD11b/CD1). J Cell Biol 1995; 130: 1473-82.
6 Giuffre L, Cordey AS, Monai N. et al. Monocyte adhesion to activated aortic endothelium: role of L-selectin and heparan sulfate proteoglycans. J Cell Biol 1997; 136: 945-56.
7 Handa K. et al. Selectin GMP-140 (CD62; PADGEM) binds to sialosyl-Le(a) and sialosyl- Le(x), and sulfated glycans modulate this binding. Biochem Biophys Res Commun 1991; 181: 1223-30.
8 Hoffmann JN, Vollmar B, Romisch J. et al. Antithrombin effects on endotoxin-induced microcirculatory disorders are mediated mainly by its interaction with microvascular endothelium. Crit Care Med 2002; 30: 218-25.
9 Inthorn D. et al. Antithrombin III supplementation in severe sepsis: beneficial effects on organ dysfunction. Shock 1997; 8: 328-34.
10 Kaneider NC. et al. Syndecan-4 mediates anti-thrombin-induced chemotaxis of human peripheral blood lymphocytes and monocytes. J Cell Sci 2002; 115: 227-36.
11 Kaneider NC. et al. Syndecan-4 as antithrom-bin receptor of human neutrophils. Biochem Biophys Res Commun 2001; 287: 42-6.
12 Dunzendorfer S, Kaneider N, Rabensteiner A. et al. Cell-surface heparan sulfate proteogly-can-mediated regulation of human neutrophil migration by the serpin antithrombin III. Blood 2001; 97: 1079-85.
13 Horie S, Ishii H, Kazama M. Heparin-like glycosaminoglycan is a receptor for antithrombin III- dependent but not for thrombin-dependent prostacyclin production in human endothelial cells. Thromb Res 1990; 59: 895-904.
14 Olson ST. et al. Role of the antithrombin-binding pentasaccharide in heparin acceleration of antithrombin-proteinase reactions. Resolution of the antithrombin conformational change contribution to heparin rate enhancement. J Biol Chem 1992; 267: 12528-38.
15 Yanada M, Kojima T, Ishiguro K. et al. Impact of antithrombin deficiency in thrombogenesis: lipopolysaccharide and stress-induced thrombus formation in heterozygous antithrombin-deficient mice. Blood 2002; 99: 2455-8.
16 Hoffmann JN. et al. Adverse effect of heparin on antithrombin action during endotoxemia: microhemodynamic and cellular mechanisms. Thromb Haemost 2002; 88: 242-52.
17 Damus PS, Rosenberg RD. Antithrombin-heparin cofactor. Methods Enzymol 1976; 45: 653-69.
18 Kaneider NC. et al. Reversal of thrombin-induced deactivation of CD39/ATPDase in endothelial cells by HMG-CoA reductase inhibition: effects on Rho- GTPase and adenosine nucleotide metabolism. Arterioscler Thromb Vasc Biol 2002; 22: 894-900.
19 Hsu HY, Wen MH. Lipopolysaccharide-mediated reactive oxygen species and signal transduction in the regulation of interleukin-1 gene expression. J Biol Chem 2002; 277: 22131-9.
20 Carrell RW. How serpins are shaping up. Science 1999; 285: 1861
21 Uchiba M, Okajima K, Murakami K. et al. Attenuation of endotoxin-induced pulmonary vascular injury by antithrombin III. Am J Physiol 1996; 270: L921-L930.
22 Okada Y, Zuo XJ, Marchevsky AM. et al. Antithrombin III treatment improves parameters of acute inflammation in a highly histoin-compatible model of rat lung allograft rejection. Transplantation 1999; 67: 526-8.
23 Oelschlager C. et al. Antithrombin III inhibits nuclear factor kappaB activation in human monocytes and vascular endothelial cells. Blood 2002; 99: 4015-20.
24 De La Cadena RA. et al. Activation of the kallikrein-kinin system after endotoxin administration to normal human volunteers. Blood 1993; 81: 3313-7.
25 Schoeffel U, Lausen M, Ruf G. et al. The overwhelming inflammatory response and the role of endotoxin in early sepsis. Prog Clin Biol Res 1989; 308: 371-6.
26 Bick RL. Disseminated intravascular coagulation: pathophysiological mechanisms and manifestations. Semin Thromb Hemost 1998; 24: 3-18.
27 Parrillo JE. Pathogenetic mechanisms of septic shock. N Engl J Med 1993; 328: 1471-7.
28 McCuskey RS, Urbaschek R, Urbaschek B. The microcirculation during endotoxemia. Cardiovasc Res 1996; 32: 752-63.
29 Lowry SF. Cytokine mediators of immunity and inflammation. Arch Surg 1993; 128: 1235-41.
30 Lam C. et al. Microvascular perfusion is impaired in a rat model of normotensive sepsis. J Clin Invest 1994; 94: 2077-83.
31 Nooteboom A, Van Der Linden CJ, Hendriks T. Tumor necrosis factor-alpha and interleukin-1beta mediate endothelial permeability induced by lipopolysaccharide-stimulated whole blood. Crit Care Med 2002; 30: 2063-8.
32 Ostrovsky L, Woodman RC, Payne D. et al. Antithrombin III prevents and rapidly reverses leukocyte recruitment in ischemia/reperfusion. Circulation 1997; 96: 2302-10.
33 Johnston B. et al. Differential roles of selectins and the alpha4-integrin in acute, subacute, and chronic leukocyte recruitment in vivo. J Immunol 1997; 159: 4514-23.
34 Granger DN, Kubes P. The microcirculation and inflammation: modulation of leukocyte-endothelial cell adhesion. J Leukoc Biol 1994; 55: 662-75.
35 Kaneider NC. et al. Sphingosine kinase-dependent directional migration of leukocytes in response to phorbol ester. Biochem Biophys Res Commun 2002; 297: 806-10.
36 Luo J, Kato M, Wang H. et al. Heparan sulfate and chondroitin sulfate proteoglycans inhibit E- selectin binding to endothelial cells. J Cell Biochem 2001; 80: 522-31.
37 Salmivirta M, Lidholt K, Lindahl U. Heparan sulfate: a piece of information. FASEB J 1996; 10: 1270-9.
38 Wadstrom T, Ljungh A. Glycosaminoglycan-binding microbial proteins in tissue adhesion and invasion: key events in microbial pathogenicity. J Med Microbiol 1999; 48: 223-33.
39 Choay J, Petitou M, Lormeau JC. et al. Structure-activity relationship in heparin: a synthetic pentasaccharide with high affinity for antithrombin III and eliciting high anti-factor Xa activity. Biochem Biophys Res Commun 1983; 116: 492-9.
40 Hull C. et al. Lipopolysaccharide signals an endothelial apoptosis pathway through TNF receptor-associated factor 6-mediated activation of c-Jun NH2-terminal kinase. J Immunol 2002; 169: 2611-8.
41 Bernfield M, Hinkes MT, Gallo RL. Developmental expression of the syndecans: possible function and regulation. Dev Suppl 1993; 205-12.
42 Gallo R. et al. Syndecans-1 and -4 are induced during wound repair of neonatal but not fetal skin. J Invest Dermatol 1996; 107: 676-83.
43 Gallo RL, Ono M, Povsic T. et al. Syndecans, cell surface heparan sulfate proteoglycans, are induced by a proline-rich antimicrobial peptide from wounds. Proc Natl Acad Sci USA 1994; 91: 11035-9.
44 Fitzgerald ML, Wang Z, Park PW. et al. Shedding of syndecan-1 and -4 ectodomains is regulated by multiple signaling pathways and mediated by a TIMP-3-sensitive metalloproteinase. J Cell Biol 2000; 148: 811-24.
45 Park PW, Pier GB, Hinkes MT. et al. Exploitation of syndecan-1 shedding by Pseudomonas aeruginosa enhances virulence. Nature 2001; 411: 98-102.
46 Park PW, Pier GB, Preston MJ. et al. Syndecan-1 shedding is enhanced by Las A, a secreted virulence factor of Pseudomonas aeruginosa. J Biol Chem 2000; 275: 3057-64.
47 Warren BL, Eid A, Singer P. et al. Caring for the critically ill patient. High-dose anti-thrombin III in severe sepsis: a randomized controlled trial. JAMA 2001; 286: 1869-78.