β2-integrins mediate stable adhesion in collisional interactions between neutrophils and ICAM-1-expressing cells

Eric Lynam, Larry A. Sklar, Andrew D. Taylor, Sriram Neelamegham, Bruce S. Edwards, C. Wayne Smith, Scott I. Simon

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

The aggregation of human neutrophils in suspension has features that are analogous to their attachment to activated endothelium in that both involve selectin and β2-integrin adhesion receptors. For the collisional interaction that forms neutrophil aggregates in suspension, there is a tethering step in which L-selectin on neutrophils binds PSGL-1. At relatively low shear rates (100200 s-1) firm adhesion is mediated in equal measure by LFA-1 binding to ICAM-3, and Mac-1 binding to an as yet undefined ligand. In this report we used a mouse melanoma cell line expressing an estimated 700,000 ICAM-1 (CD54) to examine the relative roles of LFA-1 and Mac-1 over the kinetics of heterotypic cell adhesion in shear mixed suspensions. Neither heterotypic nor homotypic neutrophil aggregates formed with application of shear alone. However, the rate of aggregation peaked within seconds of chemotactic stimulation. In contrast to homotypic aggregation, neither L- selectin nor its O-glycoprotein ligands on neutrophils contributed to heterotypic adhesion. Adhesion was inhibited in a dose-dependent manner as ICAM-1 was titrated with blocking mAb. A direct interaction between LFA-1 and ICAM-1 was preferred over the first minute of stimulation, whereas at later times adhesion was supported equally by Mac-1. Activation with MnCl2 also favored participation of the constitutively expressed LFA-1. Application of defined shear in a cone and plate viscometer showed that adhesion to the ICAM-1 cells decreased from a maximum level to baseline as shear rate increased up to 400 s-1 in a manner typical of integrin adhesion alone. In contrast, homotypic aggregation supported by the transition from selectin to integrin binding exhibited an increase in efficiency up to 800 s-1. The pathophysiological significance of receptor site density and duration of contact in collisional interactions relevant to leukocyte recruitment compared to leukocyte-endothelial cell interactions on surfaces is discussed.

Original languageEnglish (US)
Pages (from-to)622-630
Number of pages9
JournalJournal of Leukocyte Biology
Volume64
Issue number5
StatePublished - 1998
Externally publishedYes

Keywords

  • Cell adhesion
  • Flow cytometry

ASJC Scopus subject areas

  • Cell Biology

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    Lynam, E., Sklar, L. A., Taylor, A. D., Neelamegham, S., Edwards, B. S., Smith, C. W., & Simon, S. I. (1998). β2-integrins mediate stable adhesion in collisional interactions between neutrophils and ICAM-1-expressing cells. Journal of Leukocyte Biology, 64(5), 622-630.