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Updated: Jun 28, 2026

Quantitative In vitro Assay to Measure Neutrophil Adhesion to Activated Primary Human Microvascular Endothelial Cells under Static Conditions
Published on: August 23, 2013
E-selectin and intercellular adhesion molecule-1 are released by activated human endothelial cells in vitro
J F Leeuwenberg1, E F Smeets, J J Neefjes
1Department of Surgery, University of Limburg, Maastricht, The Netherlands.
Researchers discovered that endothelial cells release soluble E-selectin and ICAM-1 adhesion molecules following cytokine activation. The release kinetics differ, with E-selectin peaking early and ICAM-1 later, potentially impacting inflammatory cell interactions.
Area of Science:
- Cell Biology
- Immunology
- Biochemistry
Background:
- Endothelial cells express adhesion molecules like E-selectin and ICAM-1 upon cytokine stimulation.
- The release mechanism and fate of these cell surface molecules were previously unknown.
Purpose of the Study:
- To investigate the release of soluble E-selectin and ICAM-1 from activated human umbilical vein endothelial cells (HUVEC).
- To characterize the kinetics and correlation of soluble adhesion molecule release with cell surface expression.
Main Methods:
- Developed specific sandwich ELISAs for soluble E-selectin (sE-selectin) and soluble ICAM-1 (sICAM-1).
- Activated HUVEC with tumor necrosis factor (TNF), interleukin-1 (IL-1), or lipopolysaccharide (LPS).
- Analyzed HUVEC supernatant for soluble adhesion molecules using ELISA and biochemical methods.
Main Results:
- Soluble E-selectin and ICAM-1 were detected in HUVEC supernatant after cytokine activation.
- sE-selectin release peaked 6-12 hours post-activation and declined within 24 hours.
- sICAM-1 release increased gradually, plateauing after 24 hours and remaining stable for 3 days.
- The amount of released soluble molecules correlated directly with cell surface expression.
Conclusions:
- Endothelial cells release E-selectin and ICAM-1 into the supernatant following activation.
- The distinct release patterns of sE-selectin and sICAM-1 suggest different regulatory mechanisms.
- The physiological significance of released soluble adhesion molecules in inflammatory processes requires further investigation.
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