title: Immobilized IL-8 triggers progressive activation of neutrophils rolling in vitro on P-selectin and intercellular adhesion molecule-1.
abstract: The chemokine IL-8 is found on the luminal side of vascular endothelial cells, where it is postulated to be immobilized during inflammation. In this study, we observed that immobilized IL-8 can stimulate neutrophils to firmly adhere to a substrate containing ICAM-1 in a static adhesion assay. Soluble IL-8 was then perfused over neutrophils rolling on P-selectin (P-sel) and ICAM-1, confirming that IL-8 in solution can quickly cause rolling neutrophils to arrest. To mimic a blood vessel wall with IL-8 expressed on the luminal surface of endothelial cells, IL-8 was immobilized along with P-sel and ICAM-1 at defined site densities to a surface. Neutrophils rolled an average of 200 microm on surfaces of P-sel, ICAM-1, and IL-8 before firmly adhering through ICAM-1-beta(2) integrin interactions at 2 dynes/cm(2) wall shear stress. Increasing the density of IL-8 from 60 to 350 sites/microm(2) on the surface decreased by 50% the average distance and time the neutrophils rolled before becoming firmly adherent. Temporal dynamics of ICAM-1-beta(2) integrin interactions of rolling neutrophils following IL-8 exposure suggest the existence of two classes of beta(2) integrin-ICAM-1 interactions, a low avidity interaction with a 65% increase in pause times as compared with P-sel-P-sel glycoprotein ligand-1 interactions, and a high avidity interaction with pause times 400% greater than the selectin interactions. Based on the proportionality between IL-8 site density and time to arrest, it appears that neutrophils may need to sample a critical number of IL-8 molecules presented by the vessel wall before forming a sufficient number of high avidity beta(2) integrin bonds for firm adhesion.
authors.affiliation: Department of Biomedical Engineering, University of Virginia, Charlottesville, VA 22903, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: DiVietro
authors.forename: M J
authors.initials: MJ
authors.lastname: Smith
authors.forename: B R
authors.initials: BR
authors.lastname: Smith
authors.forename: L
authors.initials: L
authors.lastname: Petruzzelli
authors.forename: R S
authors.initials: RS
authors.lastname: Larson
authors.forename: M B
authors.initials: MB
authors.lastname: Lawrence
mesh_terms: D002448:Cell Adhesion; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D016209:Interleukin-8 / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D018715:Microscopy, Video; D018375:Neutrophil Activation; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D019007:P-Selectin / Q000378:metabolism*
chemical_list: D016209:Interleukin-8; D019007:P-Selectin; D018799:Intercellular Adhesion Molecule-1