Interplay between Rolling and Firm Adhesion Elucidated with a Cell-Free System Engineered with Two Distinct Receptor-Ligand Pairs

The firm arrest of leukocytes to the endothelium during inflammation is known to be mediated by endothelial intercellular adhesion molecules (ICAMs) binding to activated integrins displayed on leukocyte surface. Selectin-ligand interactions, which mediate rolling, are believed to be important for fa...

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Veröffentlicht in:Biophysical journal 2003-10, Vol.85 (4), p.2720-2731
Hauptverfasser: Eniola, A. Omolola, Willcox, P. Jeanene, Hammer, Daniel A.
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Willcox, P. Jeanene
Hammer, Daniel A.
description The firm arrest of leukocytes to the endothelium during inflammation is known to be mediated by endothelial intercellular adhesion molecules (ICAMs) binding to activated integrins displayed on leukocyte surface. Selectin-ligand interactions, which mediate rolling, are believed to be important for facilitating firm adhesion, either by activating integrins or by facilitating the transition to firm adhesion by making it easier for integrins to bind. Although leukocytes employ two distinct adhesion molecules that mediate different states of adhesion, the fundamental biophysical mechanisms by which two pairs of adhesion molecules facilitate cell adhesion is not well understood. In this work, we attempt to understand the interaction between two molecular systems using a cell-free system in which polystyrene microspheres functionalized with the selectin ligand, sialyl LewisX (sLeX), and an antibody against ICAM-1, aICAM-1, are perfused over P-selectin/ICAM-1 coated surfaces in a parallel plate flow chamber. Separately, sLeX/P-selectin interactions support rolling and aICAM-1/ICAM-1 interactions mediate firm adhesion. Our results show that sLeX/aICAM-1 microspheres will firmly adhere to P-selectin/ICAM-1 coated surfaces, and that the extent of firm adhesion of microspheres is dependent on wall shear stress within the flow chamber, sLeX/aICAM-1 microsphere site density, and P-selectin/ICAM-1 surface density ratio. We show that P-selectin's interaction with sLeX mechanistically facilitates firm adhesion mediated by antibody binding to ICAM-1: the extent of firm adhesion for the same concentration of aICAM-1/ICAM-1 interaction is greater when sLeX/P-selectin interactions are present. aICAM-1/ICAM-1 interactions also stabilize rolling by increasing pause times and decreasing average rolling velocities. Although aICAM-1 is a surrogate for β2-integrin, the kinetics of association between aICAM-1 and ICAM-1 is within a factor of 1.5 of activated integrin binding ICAM-1, suggesting the findings from this model system may be insightful to the mechanism of leukocyte firm adhesion. In particular, these experimental results show how two molecule systems can interact to produce an effect not achievable by either system alone, a fundamental mechanism that may pervade leukocyte adhesion biology.
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In this work, we attempt to understand the interaction between two molecular systems using a cell-free system in which polystyrene microspheres functionalized with the selectin ligand, sialyl LewisX (sLeX), and an antibody against ICAM-1, aICAM-1, are perfused over P-selectin/ICAM-1 coated surfaces in a parallel plate flow chamber. Separately, sLeX/P-selectin interactions support rolling and aICAM-1/ICAM-1 interactions mediate firm adhesion. Our results show that sLeX/aICAM-1 microspheres will firmly adhere to P-selectin/ICAM-1 coated surfaces, and that the extent of firm adhesion of microspheres is dependent on wall shear stress within the flow chamber, sLeX/aICAM-1 microsphere site density, and P-selectin/ICAM-1 surface density ratio. 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Omolola</au><au>Willcox, P. Jeanene</au><au>Hammer, Daniel A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Interplay between Rolling and Firm Adhesion Elucidated with a Cell-Free System Engineered with Two Distinct Receptor-Ligand Pairs</atitle><jtitle>Biophysical journal</jtitle><addtitle>Biophys J</addtitle><date>2003-10</date><risdate>2003</risdate><volume>85</volume><issue>4</issue><spage>2720</spage><epage>2731</epage><pages>2720-2731</pages><issn>0006-3495</issn><eissn>1542-0086</eissn><abstract>The firm arrest of leukocytes to the endothelium during inflammation is known to be mediated by endothelial intercellular adhesion molecules (ICAMs) binding to activated integrins displayed on leukocyte surface. Selectin-ligand interactions, which mediate rolling, are believed to be important for facilitating firm adhesion, either by activating integrins or by facilitating the transition to firm adhesion by making it easier for integrins to bind. 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subjects Antigen-Antibody Complex - analysis
Antigen-Antibody Complex - chemistry
Antigen-Antibody Complex - metabolism
Biochemistry
Biomimetics - methods
Cell Adhesion - physiology
Cell Biophysics
Cell Movement - physiology
Cell-Free System - chemistry
Cell-Free System - metabolism
Coated Materials, Biocompatible - chemistry
Coated Materials, Biocompatible - metabolism
Flow Cytometry - methods
Intercellular Adhesion Molecule-1 - chemistry
Intercellular Adhesion Molecule-1 - metabolism
Kinetics
Leukocytes
Microspheres
Molecular biology
Oligosaccharides - chemistry
Oligosaccharides - metabolism
P-Selectin - chemistry
P-Selectin - metabolism
Protein Binding
Rotation
Sialyl Lewis X Antigen
title Interplay between Rolling and Firm Adhesion Elucidated with a Cell-Free System Engineered with Two Distinct Receptor-Ligand Pairs
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