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1 Department of Chemical Engineering, The Johns Hopkins University, Baltimore, MD, USA
2 Department of Medicine, Division of Clinical Immunology, The Johns Hopkins University School of Medicine, Baltimore, MD, USA
* To whom correspondence should be addressed. E-mail: konst_k{at}jhu.edu.
This study examines the binding kinetics and molecular requirements of eosinophil adhesion to surface-anchored platelets in shear flow. P-selectin-glycoprotein-ligand-1 (PSGL-1) binding to platelet P-selectin initiates tethering and rolling of eosinophils to platelets under flow. These primary interacting cells assist in the capture of free-flowing eosinophils through homotypic tethering (secondary-interactions) mediated via L-selectin-PSGL-1 interactions. Differences between eosinophils and neutrophils in the PSGL-1 and L-selectin expression levels predict the pattern and relative extent of their adhesive interactions with immobilized platelets under shear, as well as the relative magnitude of their average rolling velocities. The majority of tethered eosinophils become rapidly stationary on the platelet layer, a process that is predominantly mediated via eosinophil PSGL-1 binding to platelet P-selectin, and has an absolute requirement for intact cytoskeleton. Only a small fraction of these stationary eosinophils develop shear-resistant attachments mediated by CD18 integrins. However, stimulation of eosinophils with eotaxin-2 converts PSGL-1-P-selectin-dependent stationary adhesion to CD18-mediated shear-resistant stable attachment. These studies provide insights for designing strategies to combat thrombotic disorders in hypereosinophilic patients based on blocking eosinophil-platelet interactions.
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