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Updated: Aug 8, 2026

Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions
08:54

Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions

Published on: August 8, 2012

Analysis of integrin (CD11b/CD18) movement during neutrophil adhesion and migration on endothelial cells

Y P Rochon1, T J Kavanagh, J M Harlan

  • 1Department of Molecular Biotechnology, University of Washington, Seattle, WA 98195-7710, USA.

Journal of Microscopy
|January 5, 2000
PubMed

Insights

Leukocyte adhesion molecule Mac-1 rapidly accumulates at contact points with endothelial cells and redistributes to the leading edge during neutrophil migration. This dynamic movement highlights the cell surface pool

Area of Science:

  • Immunology
  • Cell Biology
  • Microscopy

Background:

  • The distribution of cell surface molecules during leukocyte adhesion and migration on endothelial cells is not well understood.
  • Understanding these dynamics is crucial for comprehending immune cell trafficking and inflammatory responses.

Purpose of the Study:

  • To investigate the dynamic redistribution of the integrin CD11b/CD18 (Mac-1) during neutrophil adhesion and migration on human umbilical vein endothelial cells (HUVEC).
  • To assess the utility of confocal microscopy with non-inhibitory antibodies for studying cell surface molecule dynamics.

Main Methods:

  • Utilized confocal microscopy to visualize the movement of Mac-1.
  • Employed Fab fragments of a non-inhibitory monoclonal antibody against Mac-1, labeled with photostable fluorophores.
  • Observed interactions between neutrophils and TNF-alpha activated HUVEC.

Main Results:

  • Mac-1 rapidly accumulated at the initial contact area between neutrophils and HUVEC.
  • During neutrophil spreading, Mac-1 redistributed away from the initial contact zone.
  • Mac-1 was observed to move to the leading edge of migrating neutrophils on HUVEC.
  • Mac-1-dense macroaggregates formed and disassembled dynamically at the cell interface.

Conclusions:

  • The cell surface pool of Mac-1 is dynamic and can be recruited to the leading edge during neutrophil migration.
  • Confocal microscopy with labeled non-inhibitory antibodies is effective for studying the temporal dynamics of cell surface molecules.
  • These findings provide insights into the mechanisms of leukocyte-endothelial cell interactions.

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