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Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions
11:22

Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions

Published on: August 23, 2013

Adhesion of endothelial cells to NOV is mediated by the integrins alphavbeta3 and alpha5beta1

Peter D Ellis1, James C Metcalfe, Marko Hyvönen

  • 1Section of Cardiovascular Biology, University of Cambridge, Cambridge, UK. pde1001@mole.bio.cam.ac.uk

Journal of Vascular Research
|August 7, 2003
PubMed

Insights

NOV protein, a CCN family member, promotes endothelial cell adhesion to blood vessels. This interaction, crucial for vascular health and injury response, involves specific integrins and intracellular signaling pathways.

Area of Science:

  • Vascular Biology
  • Cell Adhesion
  • Extracellular Matrix Proteins

Background:

  • NOV is a matricellular protein of the CCN family, predominantly expressed by vascular smooth muscle cells (VSMCs).
  • NOV expression in rat carotid arteries is downregulated post-injury, except in the developing intima, suggesting a role in endothelial cell interactions.

Purpose of the Study:

  • To investigate the role of NOV in endothelial cell adhesion.
  • To elucidate the molecular mechanisms underlying NOV-mediated endothelial cell adhesion.

Main Methods:

  • Assessing human umbilical vein endothelial cell (HUVEC) adhesion to NOV.
  • Utilizing anti-NOV antibodies, divalent cations, GRGDS peptide, and specific integrin-blocking monoclonal antibodies (mAbs) against alphavbeta3 and alpha5beta1.
  • Analyzing intracellular signaling via phosphotyrosine content of focal adhesion kinase (FAK).

Main Results:

  • NOV significantly promoted HUVEC adhesion, an effect blocked by anti-NOV antibody.
  • HUVEC adhesion to NOV was dependent on divalent cations and inhibited by GRGDS peptide, indicating integrin involvement.
  • NOV directly bound to the alphavbeta3 integrin, and while alpha5beta1 also mediated adhesion, it required fibronectin.
  • HUVEC adhesion triggered intracellular signaling, evidenced by increased FAK phosphotyrosine levels.

Conclusions:

  • NOV plays a significant role in regulating endothelial cell adhesion.
  • The interaction involves specific integrins, notably alphavbeta3, and influences intracellular signaling pathways.
  • These findings support a role for NOV in maintaining vascular homeostasis and responding to vascular injury.

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