Platelet microparticle formation and thrombin generation under high shear are effectively suppressed by a monoclonal antibody against GPIbα

Luca Pontiggia; Beat Steiner; Hans Ulrichts; Hans Deckmyn; Marc Forestier; Jürg H. Beer

doi:10.1160/TH06-07-0367

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2006; 96(06): 774-780
DOI: 10.1160/TH06-07-0367

Platelets and Blood Cells

Schattauer GmbH

Platelet microparticle formation and thrombin generation under high shear are effectively suppressed by a monoclonal antibody against GPIbα

Luca Pontiggia

¹Department of Medicine, Laboratory for Thrombosis Research, Kantonsspital Baden, Switzerland

,

Beat Steiner

²Hoffmann-La Roche Ltd, Pharma Preclinical Research, Basel, Switzerland

,

Hans Ulrichts

³Laboratory forThrombosis Research, IRC, KU Leuven, Campus Kortrijk, Belgium

,

Hans Deckmyn

³Laboratory forThrombosis Research, IRC, KU Leuven, Campus Kortrijk, Belgium

,

Marc Forestier

¹Department of Medicine, Laboratory for Thrombosis Research, Kantonsspital Baden, Switzerland

,

Jürg H. Beer

¹Department of Medicine, Laboratory for Thrombosis Research, Kantonsspital Baden, Switzerland

› Author Affiliations

Abstract

Summary

We studied the inhibition of platelet microparticle (MP) formation and thrombin generation under high shear forces. We hypothesized that an inhibitor of the GPIbα-von Willebrand factor (vWF) interaction would be more effective in suppressing MP formation and thrombin generation than GPIIb/IIIa inhibitors. Platelet-rich plasma (PRP) anticoagulated with PPACK (D-Phe-Pro-Arg chloromethyl ketone) was exposed in a coneand-plate viscometer (shear: 5,000 s⁻¹ for5 min) in the presence of antagonists to GPIbα (the monoclonal antibody [Mab] Ib-23) or to GPIIb/IIIa (abciximab, tirofiban, eptifibatide) at their IC₉₀ determined in platelet aggregometry with ristocetin or ADP, respectively. We used double labeling (CD41-PE and annexin-V-FITC) for flow cytometric detection of MP and their aminophospholipid exposure. Thrombin generation was measured using PRP prepared fromACD anticoagulated blood. About 40% of the thrombin generation was found to be mediated by the MP fraction of the PRP. Blockade of GPIbα with Mab Ib-23 reduced MP formation and thrombin generation by 50%, and was more effective than any GPIIb/IIIa antagonist. The combination of Mab Ib-23 with one of the GPIIb/IIIa inhibitors further reduced the MP formation to ∼30%. The antibody also partially inhibited thrombin induced platelet aggregation. Epitope mapping suggested that Mab Ib-23 binds between the amino acids 201 and 268 of GPIbα, explaining the interference with vWF and thrombin interaction. In contrast to the commonly used GPIIb/IIIa antagonists, the blockade of GPIbα with Mab Ib-23 effectively reduces the prothrombotic MP generation and thrombin formation at shear rates typically found in arterial stenoses.

Keywords

Platelet microparticles - platelet glycoprotein Ib - thrombin - high shear - antiplatelet agents

Full Text

References

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