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Downregulation of the platelet surface glycoprotein Ib-IX complex in whole
blood stimulated by thrombin, adenosine diphosphate, or an in vivo wound
[see comments]
AD Michelson, PA Ellis, MR Barnard, GB Matic, AF Viles and AS Kestin
Department of Pediatrics, University of Massachusetts Medical School,
Worcester 01655.
In washed platelet systems, thrombin has been demonstrated to downregulate
the platelet surface expression of glycoprotein (GP) Ib and GPIX. In the
present study, we addressed the question as to whether, in the more
physiologic milieu of whole blood, downregulation of platelet surface GPIb
and GPIX can be induced by thrombin, adenosine diphosphate (ADP), and/or by
an in vivo wound. Thrombin-induced downregulation of GPIb and GPIX on the
surface of individual platelets in whole blood was demonstrated by the use
of flow cytometry, a panel of monoclonal antibodies (MoAbs) and, to inhibit
fibrin polymerization, the peptide glycyl-L-prolyl-L-arginyl-L-proline.
Platelets were identified in whole blood by a GPIV-specific MoAb and
exclusion of monocytes by light scattering properties. Flow cytometric
analysis of whole blood emerging from a standardized bleeding-time wound
established that downregulation of platelet surface GPIb and GPIX can occur
in vivo. A GPIb-IX complex-specific antibody indicated that the GPIb and
GPIX remaining on the surface of platelets activated in vivo or in vitro
were fully complexed. Simultaneous analysis of individual platelets by two
fluorophores demonstrated that thrombin-induced platelet surface exposure
of GMP-140 (degranulation) was nearly complete at the time that
downregulation of platelet surface GPIb-IX was initiated. However,
degranulation was not a prerequisite because ADP downregulated platelet
surface GPIb-IX without exposing GMP-140 on the platelet surface.
Inhibitory effects of cytochalasins demonstrated that the
activation-induced downregulation of both GPIX and GPIb are dependent on
actin polymerization. In summary, downregulation of the platelet surface
GPIb-IX complex occurs in whole blood stimulated by thrombin, ADP, or an in
vivo wound, and is independent of alpha granule secretion.
Volume 77,
Issue 4,
pp. 770-779,
02/15/1991
Copyright © 1991 by The American Society of Hematology
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