A juxtacrine mechanism for neutrophil adhesion on platelets involves platelet-activating factor and a selectin-dependent activation process.
The aim of this study was to identify the molecular mechanisms involved in neutrophil adhesion to immobilized platelets with particular focus on the possible existence of a juxtacrine system for neutrophil-platelet interactions. Platelets were immobilized onto collagen (type I)-coated coverslips that were placed in a flow chamber and neutrophils were perfused across these confluent monolayers at a shear stress of 1 to 4 dynes/cm2. Neutrophils rolled, and a significant proportion (25% to 50%) adhered to platelet monolayers. P-selectin was expressed in very large quantities on the surface of platelets and mediated all of the rolling, whereas the beta2-integrin mediated firm adhesion. An activation mechanism for adhesion was necessary inasmuch as fixed neutrophils continued to roll on immobilized platelets, but did not adhere. Platelets adherent to collagen produced significant levels of platelet-activating factor (PAF). Accordingly, the firm adhesion of neutrophils to platelets was significantly inhibited by a PAF receptor antagonist (WEB 2086). Treatment of only the platelets with acetylhydrolase, which converts membrane-associated PAF to lyso-PAF, prevented 60% of the adhesion. These data suggest that PAF, on the surface of platelets, mediated a significant portion of the adhesive interaction. Addition of some selectin-binding carbohydrates (fucoidan or soluble SLEx analogs but not dextran sulfate) to the platelets caused rolling neutrophils to immediately adhere, an event that was not observed on histamine or thrombin-treated endothelium or P-selectin transfectants. These data support the view that a juxtacrine activation process exists on immobilized platelets for neutrophils. This process can be greatly enhanced on platelets and may involve a signaling mechanism through P-selectin.
Ostrovsky
L
L
Department of Medical Physiology and Medicine, University of Calgary, Alberta, Canada.
King
A J
AJ
Bond
S
S
Mitchell
D
D
Lorant
D E
DE
Zimmerman
G A
GA
Larsen
R
R
Niu
X F
XF
Kubes
P
P
D000925:Anticoagulants / Q000494:pharmacology; D001792:Blood Platelets / Q000166:cytology* / Q000502:physiology*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D006801:Humans; D009504:Neutrophils / Q000166:cytology* / Q000502:physiology*; D019007:P-Selectin / Q000502:physiology*; D010972:Platelet Activating Factor / Q000502:physiology*; D015539:Platelet Activation / Q000502:physiology*; D011134:Polysaccharides / Q000494:pharmacology
D000925:Anticoagulants; D019007:P-Selectin; D010972:Platelet Activating Factor; D011134:Polysaccharides; C007789:fucoidan