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Chemoattractant-induced changes in surface expression and redistribution of
a functional ligand for P-selectin on neutrophils
M Dore, AR Burns, BJ Hughes, ML Entman and CW Smith
Departement de pathologie et microbiologie, Faculte de medecine
veterinaire, Universite de Montreal, Quebec, Canada.
Adhesion between platelets and neutrophils is mediated through the
interaction of P-selectin on activated platelets with a carbohydrate-
containing structure on neutrophils, and occurs under both static and shear
conditions. Recent studies using flow chambers have shown that neutrophils
become activated after binding to surface-adherent platelets expressing
P-selectin. The objective of the present study was to investigate the
effect of such activation on the interactions of platelet P-selectin with
its ligand on neutrophils. Flow cytometric analyses using P-selectin
chimeras revealed that activation induced a rapid and marked reduction in
chimera binding, with levels of binding decreased by 71% after 15 minutes
of stimulation with the chemotactic agent, FMLP. Using a visual assay of
platelet-neutrophil rosetting, we showed that the P-selectin ligand was
translocated and clustered at the uropod of neutrophils following the shape
changes and polarization induced by chemotactic stimulation. Activated
neutrophils bound to surface-adherent platelets also displayed the
clustering of P-selectin ligand at the uropod, and these neutrophils
detached from the platelets when a shear stress (2 dynes/cm2) was applied
through the adhesion chamber. These results indicate that chemotactic
stimulation of neutrophils induces changes in the surface expression and
distribution of a biologically relevant ligand for P-selectin, and that
these changes might influence the adhesive interactions occurring between
neutrophils and activated platelets.
Volume 87,
Issue 5,
pp. 2029-2037,
03/01/1996
Copyright © 1996 by The American Society of Hematology
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