Scientific article

Neutrophil transmigration under shear flow conditions in vitro is junctional adhesion molecule-C independent

Published inThe Journal of immunology, vol. 178, no. 9, p. 5879-5887
Publication date2007

Endothelial cell junctional adhesion molecule (JAM)-C has been proposed to regulate neutrophil migration. In the current study, we used function-blocking mAbs against human JAM-C to determine its role in human leukocyte adhesion and transendothelial cell migration under flow conditions. JAM-C surface expression in HUVEC was uniformly low, and treatment with inflammatory cytokines TNF-alpha, IL-1beta, or LPS did not increase its surface expression as assessed by FACS analysis. By immunofluorescence microscopy, JAM-C staining showed sparse localization to cell-cell junctions on resting or cytokine-activated HUVEC. Surprisingly, staining of detergent-permeabilized HUVEC revealed a large intracellular pool of JAM-C that showed little colocalization with von Willebrand factor. Adhesion studies in an in vitro flow model showed that functional blocking JAM-C mAb alone had no inhibitory effect on polymorphonuclear leukocyte (PMN) adhesion or transmigration, whereas mAb to ICAM-1 significantly reduced transmigration. Interestingly, JAM-C-blocking mAbs synergized with a combination of PECAM-1, ICAM-1, and CD99-blocking mAbs to inhibit PMN transmigration. Overexpression of JAM-C by infection with a lentivirus JAM-C GFP fusion protein did not increase adhesion or extent of transmigration of PMN or evoke a role for JAM-C in transendothelial migration. These data suggest that JAM-C has a minimal role, if any, in PMN transmigration in this model and that ICAM-1 is the preferred endothelial-expressed ligand for PMN beta(2) integrins during transendothelial migration.

  • Animals
  • Antibodies, Blocking/pharmacology
  • Antibodies, Monoclonal/pharmacology
  • Antigens, CD18/metabolism
  • Antigens, CD31/drug effects/metabolism
  • Cell Adhesion
  • Cell Adhesion Molecules/antagonists & inhibitors/immunology/ metabolism
  • Cell Movement
  • Immunoglobulins/immunology/ metabolism
  • Intercellular Adhesion Molecule-1/drug effects/metabolism
  • Interleukin-1beta/pharmacology
  • Lentivirus Infections/immunology
  • Lipopolysaccharides/pharmacology
  • Membrane Proteins/antagonists & inhibitors/immunology/ metabolism
  • Mice
  • Neutrophils/drug effects/ immunology
  • Shear Strength
  • Tumor Necrosis Factor-alpha/pharmacology
Affiliation Not a UNIGE publication
Citation (ISO format)
SIRCAR, Monica et al. Neutrophil transmigration under shear flow conditions in vitro is junctional adhesion molecule-C independent. In: The Journal of immunology, 2007, vol. 178, n° 9, p. 5879–5887. doi: 10.4049/jimmunol.178.9.5879
Updates (1)
ISSN of the journal0022-1767

Technical informations

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