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

Thromb Haemost. 2006 Dec;96(6):774-80.

Abstract

We studied the inhibition of platelet microparticle (MP) formation and thrombin generation under high shear forces. We hypothesized that an inhibitor of the GPIb a -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 cone-and-plate viscometer (shear: 5,000 s(-1) for 5 min) in the presence of antagonists to GPIb a (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 from ACD anticoagulated blood. About 40% of the thrombin generation was found to be mediated by the MP fraction of the PRP. Blockade of GPIb a 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 GPIb a , explaining the interference with vWF and thrombin interaction. In contrast to the commonly used GPIIb/IIIa antagonists, the blockade of GPIb a with Mab Ib-23 effectively reduces the prothrombotic MP generation and thrombin formation at shear rates typically found in arterial stenoses.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abciximab
  • Annexin A5 / metabolism
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / pharmacology
  • Binding Sites, Antibody
  • Blood Platelets / drug effects
  • Blood Platelets / metabolism*
  • Dose-Response Relationship, Drug
  • Epitope Mapping
  • Eptifibatide
  • Flow Cytometry
  • Humans
  • Immunoglobulin Fab Fragments / pharmacology
  • In Vitro Techniques
  • Peptides / pharmacology
  • Phosphatidylserines / metabolism
  • Platelet Aggregation Inhibitors / pharmacology
  • Platelet Aggregation* / drug effects
  • Platelet Function Tests
  • Platelet Glycoprotein GPIIb-IIIa Complex / antagonists & inhibitors
  • Platelet Glycoprotein GPIIb-IIIa Complex / metabolism*
  • Platelet Glycoprotein GPIb-IX Complex / immunology
  • Platelet Glycoprotein GPIb-IX Complex / metabolism*
  • Stress, Mechanical
  • Thrombin / metabolism*
  • Tirofiban
  • Tyrosine / analogs & derivatives
  • Tyrosine / pharmacology
  • von Willebrand Factor / metabolism

Substances

  • Annexin A5
  • Antibodies, Monoclonal
  • Immunoglobulin Fab Fragments
  • Peptides
  • Phosphatidylserines
  • Platelet Aggregation Inhibitors
  • Platelet Glycoprotein GPIIb-IIIa Complex
  • Platelet Glycoprotein GPIb-IX Complex
  • von Willebrand Factor
  • Tyrosine
  • Thrombin
  • Tirofiban
  • Eptifibatide
  • Abciximab