title: Neutrophils rolling on immobilised platelets migrate into homotypic aggregates after activation.
abstract: Interactions between platelets and leucocytes are implicated in the pathology of thrombotic vascular disease. Using a flow-based adhesion assay we have investigated a novel route for the formation of neutrophil aggregates on the surface of immobilised activated platelets. Neutrophils perfused over a platelet monolayer formed numerous rolling attachments but rapidly stopped and spread after the superfusion of N-formyl-methionyl-leucyl-phenylalanine or platelet-activating factor (both at 10(-7) M). Subsequent integrin-mediated migration across the platelet monolayer enabled formation of homotypic neutrophil aggregates, which was significant within 2.5 min of receipt of either stimulus. Aggregates increased in size with time and had an average projected area of approximately 500 microm2 after 10 min. Increasing size was correlated with an increasing tendency for movement downstream and large aggregates sometimes tumbled in that direction. The formation and stability of homotypic aggregates was dependent on several adhesive mechanisms. Antibody blockade demonstrated that interactions involving CD11a/CD18 and ICAM-3, between alpha(v)beta3-integrin and CD31 and between L-selectin and an unidentified counter-ligand were all required for the complete aggregatory response. Furthermore, blockade of L-selectin allowed initial aggregation which then reversed, suggesting that this receptor might regulate the interactions between other adhesion molecules that directly supported cell-cell adhesion. We propose that this novel route for leucocyte aggregation could promote vascular occlusion in thrombotic vessels or at distal sites in the event of embolisation.
authors.affiliation: Department of Physiology, The Medical School, The University of Birmingham, UK. raingege@bham.ac.uk
authors.forename: G E
authors.initials: GE
authors.lastname: Rainger
authors.forename: C
authors.initials: C
authors.lastname: Buckley
authors.forename: D L
authors.initials: DL
authors.lastname: Simmons
authors.forename: G B
authors.initials: GB
authors.lastname: Nash
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD; D000943:Antigens, Differentiation; D001792:Blood Platelets; D018821:CD18 Antigens / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000502:physiology*; D002449:Cell Aggregation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D048429:Cell Size; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D018056:Hemorheology; D006801:Humans; D016023:Integrins / Q000502:physiology; D019041:L-Selectin / Q000502:physiology; D016177:Macrophage-1 Antigen / Q000502:physiology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D010972:Platelet Activating Factor / Q000494:pharmacology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000502:physiology; D019030:Receptors, Vitronectin / Q000502:physiology
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000943:Antigens, Differentiation; D018821:CD18 Antigens; D015815:Cell Adhesion Molecules; C483616:ICAM3 protein, human; D016023:Integrins; D016177:Macrophage-1 Antigen; D010972:Platelet Activating Factor; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D019030:Receptors, Vitronectin; D019041:L-Selectin; D009240:N-Formylmethionine Leucyl-Phenylalanine