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 αIIbβ3. 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 αIIbβ3–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 αIIbβ3 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 αIIbβ3 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 αIIbβ3 and, in a shear-dependent way, on GP Ib.
Keywords:
FITC (fluorescein isothiocyanate), GP Ib-IX-V (glycoprotein Ib-IX-V), HEPES (N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid), PAR1 (protease-activated receptor 1), PFP (platelet-free plasma), PRP (platelet-rich plasma), vWF (von Willebrand factor), Z-GGR-AMC (benzyloxycarbonyl Gly-Gly-Arg 7-amido-4-methyl-coumarin)To read this article in full you will need to make a payment
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Article info
Publication history
Accepted:
November 26,
2002
Received:
October 25,
2002
Identification
Copyright
© 2003 Elsevier Science Inc. Published by Elsevier Inc. All rights reserved.
