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Blood, Vol. 93 No. 11 (June 1), 1999:
pp. 3831-3838
Endothelial Cells Undergoing Apoptosis Become Proadhesive for
Nonactivated Platelets
Thomas Bombeli,
Barbara R. Schwartz, and
John M. Harlan
From the Division of Hematology, University of
Washington, Seattle, WA; and the Division of Hematology, the Department
of Medicine, University Hospital of Zurich, Zurich, Switzerland.
Under normal conditions, platelets do not adhere to endothelium.
However, when platelets or endothelial cells are stimulated by thrombin
or cytokines, respectively, platelets bind avidly to endothelium.
Because there is accumulating evidence that endothelial cells may
become apoptotic under certain proinflammatory or prothrombotic conditions, we investigated whether endothelial cells undergoing apoptosis may become proadhesive for nonactivated platelets. Human umbilical vein endothelial cells (HUVEC) were induced to undergo apoptosis by staurosporine, a nonspecific protein kinase inhibitor, or
by culture in suspension with serum-deprivation. After treatment of
HUVEC or platelets with different receptor antagonists, nonactivated, washed human platelets were allowed to adhere to HUVEC for 20 minutes.
To exclude matrix involvement, platelet binding was measured in
suspension by using flow cytometry. Independent of the method of
apoptosis induction, there was a marked increase in platelet binding to
apoptotic HUVEC. Although HUVEC exhibited maximal adhesiveness for
platelets after 2 to 4 hours, complete DNA fragmentation of HUVEC
occurred only several hours later. Adhesion assays after blockade of
different platelet receptors showed only involvement of
 1-integrins. Platelet binding to apoptotic HUVEC was
inhibited by more than 70% when platelets were treated with blocking
anti-1 antibodies. Treatment of apoptotic HUVEC with
blocking antibodies to different potential platelet receptors,
including known ligands for 1-integrins, did not affect
platelet binding. As assessed by determination of -thromboglobulin
and platelet factor 4 in the supernatants, platelets bound to apoptotic
HUVEC became slightly activated. However, significant expression of
platelet P-selectin (CD62P) was not found. These data provide further
evidence that endothelial cells undergoing apoptosis may contribute to
thrombotic events.
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