title: Neutrophils can adhere via alpha4beta1-integrin under flow conditions.
abstract: In this study we investigated the possibility that an alternative pathway exists for neutrophil recruitment, namely an alpha4beta1-dependent pathway. A parallel plate chamber was used to investigate whether neutrophils could tether, roll, and adhere to tumor necrosis factor alpha (TNF alpha)-stimulated endothelium via alpha4beta1. alpha4beta1-integrin was induced on neutrophils using dihydrocytochalasin B and either an endogenous (endothelial-derived) chemotactic agent or an exogenous chemotactic molecule. alpha4beta1-expressing neutrophils could stably adhere under shear force (2 dyne/cm2) to TNF alpha-stimulated endothelium independent of the beta2-integrin. The firm adhesion was entirely abolished by antibodies directed against either the alpha4 or beta1-integrin subunits. However, the rolling interaction was not dependent on alpha4beta1 but was abolished by antiselectin therapy. Neutrophils expressing alpha4beta1 could also tether to the endothelium in the presence of antiselectin therapy, but at shear stresses less than 2 dyne/cm2. alpha4beta1-expressing neutrophils also tethered to and stably adhered (no rolling) to VCAM-1- but not to ICAM-1-transfected L cells. The interaction only occurred at shear stress less than 2 dyne/cm2. A cell line (Ramos) known to express high quantities of alpha4beta1-integrin interacted with VCAM-1-transfected L cells at very similar shear conditions. alpha4beta1-expressing neutrophils were also able to adhere to a second alpha4-integrin ligand, fibronectin; however, this interaction only occurred under static conditions. These data suggest that, under certain conditions, neutrophils can adhere independently of the beta2-integrin pathway and adhere via the alpha4beta1-integrin. This study refutes the concept that alpha4beta1-integrin adhesion is restricted to mononuclear leukocytes and is not functional on human neutrophils.
authors.affiliation: Immunology Research Group, University of Calgary, Alberta, Canada.
authors.forename: P H
authors.initials: PH
authors.lastname: Reinhardt
authors.forename: J F
authors.initials: JF
authors.lastname: Elliott
authors.forename: P
authors.initials: P
authors.lastname: Kubes
mesh_terms: D000818:Animals; D018821:CD18 Antigens / Q000502:physiology; D002448:Cell Adhesion; D002478:Cells, Cultured; D002630:Chemotactic Factors / Q000494:pharmacology; D003571:Cytochalasin B / Q000031:analogs & derivatives / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D039041:Integrin alpha4beta1; D016023:Integrins / Q000502:physiology*; D007739:L Cells; D007964:Leukocytosis; D051379:Mice; D009504:Neutrophils / Q000166:cytology* / Q000187:drug effects; D016201:Receptors, Lymphocyte Homing / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012212:Rheology; D013314:Stress, Mechanical; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D018821:CD18 Antigens; D002630:Chemotactic Factors; D005353:Fibronectins; D039041:Integrin alpha4beta1; D016023:Integrins; D016201:Receptors, Lymphocyte Homing; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; C024794:dihydrocytochalasin B; D003571:Cytochalasin B