title: Immobilized IL-8 Triggers Phagocytosis and Dynamic Changes in Membrane Microtopology in Human Neutrophils.
abstract: The interaction of leukocytes with surface bound ligands can be limited by the location of the molecules relative to the surface topology of the cell. In this report, we examine the dynamic response of neutrophils to IL-8-fractalkine chimera immobilized on bead surfaces, taking into account changes in receptor occupancy resulting from changes in surface topography. As a readout for receptor signaling, we observe the dynamics of calcium release in neutrophils following contact with the IL-8 coated surface. After a delay that depended on the initial area of contact and the surface density of IL-8, the cell began to phagocytose the IL-8 coated bead. This appeared to be a pre-requisite for release of calcium, which typically followed shortly after the initiation of phagocytosis. In separate experiments, effective kinetic coefficients for the formation of bonds between immobilized IL-8 and receptors on the cell surface were determined. Using these coefficients, we were able to estimate the number of bound receptors in the nascent contact zone. Kinetic modeling of the signaling response predicted that cell spreading and a concomitant increase in the density of occupied receptors would be required for the experimentally observed calcium dynamics. Postulating that there is an increase in receptor occupancy resulting from smoothing of the cell surface as it is stretched over the bead enabled us to obtain model predictions consistent with experimental observations. This study reveals the likely importance of membrane microtopology as a rate-limiting property and potential means of regulation of cell responses stimulated by two-dimensional surface interactions.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Pennsylvania, 240 Skirkanich Hall, 210 S. 33rd St., Philadelphia, PA, 19104, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Beste
authors.forename: Elena B
authors.initials: EB
authors.lastname: Lomakina
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Hammer
authors.forename: Richard E
authors.initials: RE
authors.lastname: Waugh
mesh_terms: D020013:Calcium Signaling; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D016209:Interleukin-8 / Q000737:chemistry*; D008954:Models, Biological; D009504:Neutrophils / Q000378:metabolism*; D010587:Phagocytosis
chemical_list: C508606:CXCL8 protein, human; D055767:Immobilized Proteins; D016209:Interleukin-8