Hammer, Daniel A

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Now showing 1 - 10 of 25
  • Publication
    Multiparticle Adhesive Dynamics. Interactions between Stably Rolling Cells
    (2001-08-01) King, Michael R; Hammer, Daniel A
    A novel numerical simulation of adhesive particles (cells) reversibly interacting with an adhesive surface under flow is presented. Particle–particle and particle–wall hydrodynamic interactions in low Reynolds number Couette flow are calculated using a boundary element method that solves an integral representation of the Stokes equation. Molecular bonds between surfaces are modeled as linear springs and stochastically formed and broken according to postulated descriptions of force-dependent kinetics. The resulting simulation, Multiparticle Adhesive Dynamics, is applied to the problem of selectin-mediated rolling of hard spheres coated with leukocyte adhesion molecules (cell-free system). Simulation results are compared to flow chamber experiments performed with carbohydrate-coated spherical beads rolling on P-selectin. Good agreement is found between theory and experiment, with the main observation being a decrease in rolling velocity with increasing concentration of rolling cells or increasing proximity between rolling cells. Pause times are found to increase and deviation motion is found to decrease as pairs of rolling cells become closer together or align with the flow.
  • Publication
    Adhesive Dynamics Simulation of Neutrophil Arrest with Deterministic Activation
    (2006-08-01) Krasik, Ellen F; Yee, Ka Lai; Hammer, Daniel A
    The transition from rolling to firm adhesion is a key element of neutrophil activation and essential to the inflammatory response. Although the molecular mediators of rolling and firm adhesion are known to be selectins and ß2-integrins, respectively, the precise dynamic mechanism by which these ligands facilitate neutrophil arrest remains unknown. Recently, it has been shown that ligation of E-selectin can stimulate the firm adhesion of neutrophils via a MAP-kinase cascade. To study the possible mechanism by which neutrophil arrest could occur, we created an integrated model by combining two methodologies from computational biology: a mechanics-based modeling of leukocyte adhesion (adhesive dynamics) and signal transduction pathway modeling. Within adhesive dynamics, a computational method our group has shown to accurately recreate rolling dynamics, we include a generic, tunable integrin activation module that links selectin engagement to integrin and activity. This model allows us to relate properties of the activation function to the dynamics of rolling and the time and distance rolled before arrest. This integrated model allows us to understand how intracellular signaling activity can set the timescale of neutrophil activation, adhesion, and diapedesis.
  • Publication
    Sialyl LewisX-Mediated, PSGL-1-Independent Rolling Adhesion on P-selectin
    (2000-08-01) Rodgers, Stephen D; Camphausen, Raymond T; Hammer, Daniel A
    Selectin-mediated cell adhesion is an essential component of the inflammatory response. In an attempt to unambiguously identify molecular features of ligands that are necessary to support rolling adhesion on P-selectin, we have used a reconstituted (“cell-free”) system in which ligand-coated beads are perfused over soluble P-selectin surfaces. We find that beads coated with the saccharides sialyl LewisX (sLeX), sialyl Lewisa (sLea), and sulfated LewisX (HSO3LeX) support rolling adhesion on P-selectin surfaces. Although it has been suggested that glycosylation and sulfation of P-selectin glycoprotein ligand-1 (PSGL-1) is required for high-affinity binding and rolling on P-selectin, our findings indicate that sulfation of N-terminal tyrosine residues is not required for binding or rolling. However, beads coated with a tyrosine-sulfated, sLeX-modified, PSGL-1-Fc chimera support slower rolling on P-selectin than beads coated with sLeX alone, suggesting that sulfation improves rolling adhesion by modulating binding to P-selectin. In addition, we find it is not necessary that P-selectin carbohydrate ligands be multivalent for robust rolling to occur. Our results demonstrate that beads coated with monovalent sLeX, exhibiting a more sparse distribution of carbohydrate than a similar amount of the multivalent form, are sufficient to yield rolling adhesion. The relative abilities of various ligands to support rolling on P-selectin are quantitatively examined among themselves and in comparison to human neutrophils. Using stop-time distributions, rolling dynamics at video frame rate resolution, and the average and variance of the rolling velocity, we find that P-selectin ligands display the following quantitative trend, in order of decreasing ability to support rolling adhesion on P-selectin: PSGL-1-Fc > sLea ~ sLeX > HSO3>LeX.
  • Publication
    HIV Viral Docking: Model Predictions for Bond Number and Trajectory
    (2003-03-22) English, Thomas J.; Hammer, Daniel A
    Viruses are nano-scale pathogenic particles. Understanding viral attachment is important to understand infectivity, disease transmission, and virus propagation throughout the host. A new simulation technique has been developed to study viral docking behavior - Brownian Adhesive Dynamics (BRAD). BRAD couples Brownian motion algorithm with adhesive dynamic models, and incorporates the effect of virus/cell geometry - an improvement over previous models. The method is extendable to any virus/cell system as well as nanoparticle adhesion system. Current studies have focused on the HIV/CD4 cell system. Comparison of BRAD simulation predictions with those of previous models of viral ducking has shown differences in steady state bond number and bond trajectory. This indicates that geometry of the system plays a significant role in the bonding behavior of viruses. Thus, it is shown that the equivalent site hypothesis is suspect.
  • Publication
    The State Diagram for Cell Adhesion Mediated by Two Receptors
    (2003-04-01) Bhatia, Sujata K; King, Michael R; Hammer, Daniel A
    Leukocyte recruitment from the bloodstream to surrounding tissues is an essential component of the immune response. Capture of blood-borne leukocytes onto vascular endothelium proceeds via a two-step mechanism, with each step mediated by a distinct receptor-ligand pair. Cells first transiently adhere, or "roll" (via interactions between selectins and sialyl- Lewis-x), and then firmly adhere to the vascular wall (via interactions between integrins and ICAM-1). We have reported that a computational method called adhesive dynamics (AD) accurately reproduces the fine-scale dynamics of selectin-mediated rolling. This paper extends the use of AD simulations to model the dynamics of cell adhesion when two classes of receptors are simultaneously active: one class (selectins or selectin ligands) with weakly adhesive properties, and the other (integrins) with strongly adhesive properties. AD simulations predict synergistic functions of the two receptors in mediating adhesion. At a fixed density of surface ICAM-1, increasing selectin densities lead to greater pause times and an increased tendency toward firm adhesion; thus, selectins mechanistically facilitate firm adhesion mediated by integrins. Conversely, at a fixed density of surface selectin, increasing ICAM-1 densities lead to greater pause times and an increased tendency to firm adhesion. We present this relationship in a two-receptor state diagram, a map that relates the densities and properties of adhesion molecules to various adhesive behaviors that they code, such as rolling or firm adhesion. We also present a state diagram for neutrophil activation, which relates β2-integrin density and integrin-ICAM-1 kinetic on rate to neutrophil adhesive behavior. The predictions of two-receptor adhesive dynamics are validated by the ability of the model to reproduce in vivo neutrophil rolling velocities from the literature.
  • Publication
    Adhesion of Polymer Vesicles
    (2005-07-06) Lin, John J.; Bates, Frank S.; Hammer, Daniel A; Silas, James A.
    The adhesion and bending modulus of polybutadiene-poly(ethylene oxide) block copolymer vesicles made from a bidisperse mixture of polymers is measured using micropipette aspiration. The adhesion energy between biotinylated vesicles and avidin beads is modeled by incorporating the extension of the adhesive ligands above the surface brush of the vesicle according to the blob model of bidisperse polymer mixtures of Komura and Safran assuming the polymer brush at the surface of the vesicle is compact. The same model accurately reproduces the scaling of the bending modulus with polymer composition.
  • Publication
    Effect of Shear Stress on Platelet Activation via the Glycoprotein VI Receptor
    (2002-10-23) Sarratt, Kendra L.; Chen, Hong M.; Kahn, Mark L.; Hammer, Daniel A
    Cardiovascular diseases are the nation's leading cause of death. Such diseases are caused by platelet response to collagen especially in the event of vascular injury leading to thrombosis. One of the platelet receptors known to bind to the collagen ligand is glycoprotein VI (GPVI) with co-receptor Fc receptor γ chain (FcRγ). By stably expressing the GPVI receptor in rat basophilic leukemia cells (RBL-2H3), which abundantly express FcRγ, but endogenously lack GPVI, studies have shown that GPVI-FcRγ is sufficient to confer adhesion as well as signaling responses to collagen as long as the receptor density is equivalent to that found on human platelets. While those investigations confirm that the GPVI receptor mediate binding to collagen under static conditions, they do not provide information on how the GPVI receptor interacts with collagen under dynamic conditions. In the present study we have used the GPVI-expressing RBL-2H3 cells to observe the kinetics of adhesion to collagen under hydrodynamic flow conditions in vitro using a parallel plate flow chamber coupled with video microscopy. We demonstrate that these cells do adhere to the surface at a low shear rate and do so at a greater adherent cell density than wild-type RBL-2H3 (WT-RBL) cells.
  • Publication
    A Microcantilever Device to Assess the Effect of Force on the Lifetime of Selectin-Carbohydrate Bonds
    (2001-02-01) Tees, Davis F. J.; Waugh, Richard E; Hammer, Daniel A
    A microcantilever technique was used to apply force to receptor-ligand molecules involved in leukocyte rolling on blood vessel walls. E-selectin was adsorbed onto 3-μm-diameter, 4-mm-long glass fibers, and the selectin ligand, sialyl Lewisx, was coupled to latex microspheres. After binding, the microsphere and bound fiber were retracted using a computerized loading protocol that combines hydrodynamic and Hookean forces on the fiber to produce a range of force loading rates (force/time), rf. From the distribution of forces at failure, the average force was determined and plotted as a function of ln rf. The slope and intercept of the plot yield the unstressed reverse reaction rate, kro , and a parameter that describes the force dependence of reverse reaction rates, ro. The ligand was titrated so adhesion occurred in ~30% of tests, implying that >80% of adhesive events involve single bonds. Monte Carlo simulations show that this level of multiple bonding has little effect on parameter estimation. The estimates are ro = 0.048 and 0.016 nm and kro = 0.72 and 2.2 s-1 for loading rates in the ranges 200–1000 and 1000–5000 pN s-1, respectively. Levenberg-Marquardt fitting across all values of rf gives ro = 0.034 nm and kro = 0.82 s-1. The values of these parameters are in the range required for rolling, as suggested by adhesive dynamics simulations.
  • Publication
    Rolling Adhesion of Yeast Engineered to Express Cell Adhesion Molecules
    (2002-04-20) Bhatia, Sujata K; Swers, Jeffrey S.; Camphausen, Raymond T.; Hammer, Daniel A; Wittrup, K. Dane
    Selectins are cell adhesion molecules that mediate capture of leukocytes on vascular endothelium as an essential component of the inflammatory response. Here we describe a method for yeast surface display of selectins, together with a functional assay that measures rolling adhesion of selectin-expressing yeast on a ligand-coated surface. E-selectin-expressing yeast roll specifically on surfaces bearing sialyl-Lewisx ligands. Observation of yeast rolling dynamics at various stages of their life cycle indicates that the kinematics of yeast motion depends on the ratio of the bud radius to the parent radius (B/P). Large-budded yeast "walk" across the surface, alternately pivoting about bud and parent. Small-budded yeast "wobble" across the surface, with bud pivoting about parent. Tracking the bud location of budding yeast allows measurement of the angular velocity of the yeast particle. Comparison of translational and angular velocities of budding yeast demonstrates that selectin-expressing cells are rolling rather than slipping across ligand-coated surfaces.
  • Publication
    The Shear Threshold Effect for Particle Adhesion to Bioreactive Surfaces: Influence of Receptor and Ligand Site Density
    (2003-03-22) Bhatia, Sujata K.; Hammer, Daniel A
    Selectins are cell adhesion molecules that mediate capture and rolling adhesion of white blood cells to vascular walls, an essential component of the inflammatory response. Adhesion through L-selectin requires a hydrodynamic shear stress above a threshold level, a phenomenon known as the shear threshold effect. We have reported that the shear threshold effect can he re-created in cell-free systems, in which microspheres coated with the carbohydrate ligand sialyl Lewis x (sLex) are perfused over L-selectin-coated surfaces. This paper extends the use of the cell-free system to determine the concurrent influence of receptor and ligand site density on the shear threshold effect. We find that the shear threshold effect diminishes with increasing levels of either L-selectin or sLex. At reduced site densities of either L-selectin or sLex, the shear threshold effect is present, with maximal rolling observed at a shear stress of 1.2 dynes/cm2. At higher site densities of L-selectin and sLex, the shear threshold effect disappears. These results suggest that the shear threshold relies on the formation of low numbers of receptor-ligand bonds.