Hydrodynamic shear and tethering through E-selectin signals phosphorylation of p38 MAP kinase and adhesion of human neutrophils

Eric Hentzen, Daniel McDonough, Larry McIntire, C. Wayne Smith, Harry L. Goldsmith, Scott I. Simon

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

Recently, we reported that tethering and rolling of neutrophils in shear flow over a substrate of E-selectin signals activation of β2-integrins and firm adhesion via an intracelullar signaling pathway involving phosphorylation of p38 MAP kinase. In the current study the objective was to examine the molecular mechanisms and shear dependence underlying activation and adhesion of β2-integrin during shear-induced collisions between human neutrophils and murine B cells (300.19) transfected to express either E-selectin or L-selectin. Three separate parameters of cell activation were assessed over the time course of application of a defined shear field to heterotypic cell suspensions in a cone-plate viscometer. These were the two-body collision doublet lifetime and capture efficiency, surface upregulation of CD11b/CD18, and tyrosine phosphorylation of p38 MAP kinase. The data indicate that neutrophil adhesion to E-selectin expressing 300.19 cells occurs with a fourfold higher efficiency of firm adhesion than do collisions with L-selectin or parent control cells. Visual analysis of aggregation in a transparent cone-plate rheoscope revealed that the lifetime and efficiency of doublet formation increased four-fold as the applied shear stress increased. Neutrophil tethering via E-selectin was associated with rapid activation as indicated by upregulation of surface CD11b/CD18 and phosphorylation of p38 MAP kinase within seconds of application of shear. Activation greatly exceeded that observed for neutrophils sheared alone or with B cells expressing L-selectin. A distinct dependence of activation on the magnitude of the shear rate suggests a coupling between the fluid mechanical effects of shear and signaling of neutrophil adhesion.

Original languageEnglish (US)
Pages (from-to)987-1001
Number of pages15
JournalAnnals of Biomedical Engineering
Volume30
Issue number8
DOIs
Publication statusPublished - 2002

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Keywords

  • Cell adhesion
  • Fluid shear
  • Integrin
  • MAP kinase
  • Neutrophilis
  • Selectin

ASJC Scopus subject areas

  • Biomedical Engineering

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