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Molecular requirements for the interaction of thrombospondin with
thrombin-activated human platelets: modulation of platelet aggregation
C Legrand, V Thibert, V Dubernard, B Begault and J Lawler
Unite 150 INSERM, Hopital Lariboisiere/Hopital St Louis, Paris, France.
We have investigated the molecular requirements for thrombospondin (TSP) to
bind to the platelet surface and to support the subsequent
secretion-dependent platelet aggregation. For this, we used two distinct
murine monoclonal antibodies (MoAbs), designated MAI and MAII, raised
against human platelet TSP, and three polyclonal antibodies, designated R3,
R6, and R5, directed against fusion proteins containing the type 1 (Gly
385-Ile 522), type 2 (Pro 559-Ile 669), and type 3 (Asp 784-Val 932)
repeating sequences, respectively. Among them, R5 and R6, but not R3,
inhibited thrombin-induced aggregation of washed platelets and the
concomitant secretion of serotonin. These antibodies, however, did not
inhibit the expression of TSP on thrombin-activated platelets, as measured
by the binding of a radiolabeled MoAb to TSP, suggesting that they may
inhibit platelet aggregation by interfering with a physiologic event
subsequent to TSP binding. In contrast, MoAb MAII, which reacts with an
epitope located within the heparin-binding domain of TSP, inhibited both
TSP surface expression and platelet aggregation/secretion induced by
thrombin. In addition, this MoAb inhibited in a dose-dependent manner (IC50
approximately 0.5 mumol/L) the interaction of 125I-TSP with immobilized
fibrinogen and platelet glycoprotein IV, both potential physiologic
receptors for TSP on thrombin-activated platelets. These results indicate
that the interaction of TSP with the surface of activated platelets can be
modulated at the level of a specific epitope located within the amino
terminal heparin-binding domain of the molecule. Thus, selective inhibition
of the platelet/TSP interaction may represent an alternative approach to
the inhibition of platelet aggregation.
Volume 79,
Issue 8,
pp. 1995-2003,
04/15/1992
Copyright © 1992 by The American Society of Hematology
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