Integrin α IIbβ 3 and shear-dependent action of glycoprotein ibα stimulate platelet-dependent thrombin formation in stirred plasma

Abstract

Under conditions of arterial-wall shear rates, platelets bind to von Willebrand factor (vWf) by way of the glycoprotein Ib (GP Ib) complex and integrin α IIbβ 3. Both adhesive receptors may also play roles in the development of procoagulant activity of platelets. Here, we investigated the effect of shear stress, as provided by a rotating cylinder, on GP Ib– and integrin α IIbβ 3–dependent thrombin generation in coagulating platelet-rich plasma (PRP). We measured thrombin continuously with the use of fluorometry from the cleavage rate of a fluorescent low-affinity substrate. The integrin α IIbβ 3 antagonist abciximab progressively reduced the peak of thrombin formation up to 43% when rate of stirring and shear stress were increased (estimated shear rates of 105-420 s −1). Abciximab did not lower the peak of thrombin formation in stirred PRP from patients with Glanzmann’s thrombasthenia lacking α IIbβ 3 but, surprisingly, shortened the time until onset. In PRP from control subjects, antibodies specifically directed against vWf-binding epitopes on GP Ibα reduced thrombin formation, with 25% to 30% at the high but not at the low stirring rate. In combination with the anti–GP Ib antibody, abciximab retained its strong inhibitory effect only at the high stirring rate. We conclude that thrombin formation and coagulation in stirred PRP depend, to a large extent, on platelet adhesion to integrin α IIbβ 3 and, in a shear-dependent way, on GP Ib.