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Platelet-dependent primary hemostasis promotes selectin- and integrin-
mediated neutrophil adhesion to damaged endothelium under flow conditions
PH Kuijper, HI Gallardo Torres, JA van der Linden, JW Lammers, JJ Sixma, L Koenderman and JJ Zwaginga
Department of Haematology, University Hospital Utrecht, The Netherlands.
Co-localization of blood platelets and granulocytes at sites of hemostasis
and inflammation has triggered an intense interest in possible interactions
between these cellular processes and induction of vessel wall injury.
Leukocyte adhesion to endothelial cells decreases with increasing shear and
is dependent on an initial rolling phase mediated by selectins. We
hypothesized that flow-dependent platelet adhesion at an injured vessel
wall will lead to P-selectin expression by platelets, thus mediating
leukocyte co-localization. A perfusion chamber was used in which flowing
whole blood induced platelet adhesion to a subendothelial matrix (ECM) of
cultured human umbilical vein endothelial cells (HUVEC). We compared
neutrophil (polymorphonuclear leukocyte [PMN]) interactions with HUVEC and
their ECM with and without adhered platelets. PMNs adhered predominantly to
ECM-adhered platelets and not to endothelial cells. ECM alone did not
support PMN adhesion under flow conditions. PMN adhesion to unstimulated
HUVEC was only substantial at low shear (up to 200 cells/mm2 at shear
stress 80 mPa). In marked contrast, PMN adhesion to ECM-adhered platelets
was dramatically increased, and adhesion was demonstrated at much higher
shear stress (up to 640 mPa). Studies with specific antibodies showed that
the platelet-dependent neutrophil adhesion was selectin-mediated.
Inhibition of P-selectin caused a marked inhibition of adhesion at high
shear stress, whereas the role of leukocyte L-selectin was less pronounced.
beta2-Integrin-blocking antibodies inhibited static neutrophil adhesion.
fMLP induced L-selectin shedding from leukocytes, resulting in decreased
leukocyte adhesion. In conclusion, platelet- dependent hemostasis at the
ECM appears to be a powerful intermediate in neutrophil-vessel wall
interactions at shear stresses that normally do not allow neutrophil
adhesion to intact endothelium.
Volume 87,
Issue 8,
pp. 3271-3281,
04/15/1996
Copyright © 1996 by The American Society of Hematology
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