Therapeutic immunoglobulin reduces Ca2+ mobilization and von Willebrand factor secretion, and increases nitric oxide release in human endothelial cells

Monique David-Dufilho; Olivier Schussler; Marie-Gabrielle Pernollet; Annie Brunet; Elisabeth Millanvoye-Van Brussel; Kim Hahn Le Quan Sang; Francine Rendu

doi:10.1160/TH03-03-0186

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 90(06): 1046-1053
DOI: 10.1160/TH03-03-0186

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Therapeutic immunoglobulin reduces Ca²⁺ mobilization and von Willebrand factor secretion, and increases nitric oxide release in human endothelial cells

Monique David-Dufilho

²Research unit “Cellular signalling and early atherosclerosis”, CNRS UMR 7131, Hôpital Broussais, Hôpital Européen George Pompidou, Paris, France

,

Olivier Schussler

¹Service de chirurgie cardiaque, Hôpital Européen George Pompidou, Paris, France

,

Marie-Gabrielle Pernollet

²Research unit “Cellular signalling and early atherosclerosis”, CNRS UMR 7131, Hôpital Broussais, Hôpital Européen George Pompidou, Paris, France

,

Annie Brunet

²Research unit “Cellular signalling and early atherosclerosis”, CNRS UMR 7131, Hôpital Broussais, Hôpital Européen George Pompidou, Paris, France

,

Elisabeth Millanvoye-Van Brussel

²Research unit “Cellular signalling and early atherosclerosis”, CNRS UMR 7131, Hôpital Broussais, Hôpital Européen George Pompidou, Paris, France

,

Kim Hahn Le Quan Sang

²Research unit “Cellular signalling and early atherosclerosis”, CNRS UMR 7131, Hôpital Broussais, Hôpital Européen George Pompidou, Paris, France

,

Francine Rendu

²Research unit “Cellular signalling and early atherosclerosis”, CNRS UMR 7131, Hôpital Broussais, Hôpital Européen George Pompidou, Paris, France

› Author Affiliations

Abstract

Summary

Intravenous γ-immunoglobulin (IVIg) is commonly used in the treatment of autoimmune and inflammatory vascular disorders to prevent thrombotic complications. The mechanism of action of IVIg is, however, not yet elucidated. In view of this, we investigated the ability of IVIg to modulate i) Ca²⁺ signals of fura-2 loaded endothelial cells, and ii) the associated release of nitric oxide (NO) and von Willebrand factor (vWf). NO was measured either indirectly by radioimmunoassay of cGMP in unstimulated cells or directly by electrochemistry at the surface of stimulated endothelial cells from human umbilical cord veins (HUVEC). Short-term treatment of unstimulated HUVEC with intact IVIg decreased the basal cytosolic Ca²⁺ concentration by 20% while it activated the NO/cGMP synthesis. Following IVIg treatment of HUVEC, the Ca²⁺ liberation from internal stores and the vWf secretion induced by ATP, thrombin or histamine were significantly reduced by 38 and 60 %, respectively. The effects on Ca²⁺ signals were observed with intact IVIg as well as with the F(ab’)2 or the Fc fragments indicating that both portions are involved in the mechanism of action. The IVIg treatment of HUVECs had no effect on the NO release induced by thrombin or histamine. By contrast, the IVIg treatment increased the ATP-activated NO release by amplifying the Ser1177-eNOS phosphorylation. The IVIg also activated the NO-dependent cGMP release in resting and collagen-stimulated platelets. Since NO is a potent inhibitor of platelet activation and vWF is a platelet adhesion cofactor, the beneficial effects of therapeutic IVIg may lie in the inhibition of platelet adhesion to damaged endothelium.

Keywords

Endothelial cells - IVIg - nitric oxide - platelets - von Willebrand factor

Full Text

References

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