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Updated: Aug 6, 2026
Quantitation of Endothelial Cell Adhesiveness In Vitro
Published on: June 18, 2015
Enhanced adhesion of polymorphonuclear leukocytes to anoxic cultured vascular endothelium
J M Budd1, S J Kirtland, H Baum
1Roche Products Ltd., Welwyn Garden City, Hertfordshire, U.K.
Abstract:
This study showed that the adherence of human polymorphonuclear leukocytes (PMN) to monolayer cultures of human umbilical vein endothelial cells (HUVEC) was increased when the latter were rendered anoxic. This adhesion was greater after 4-5 h than after 8 h of anoxia, but even at 8 h was significantly above the level of adhesion to HUVEC maintained under normoxic conditions for the same period. The changes in adhesion were not dependent on the viability of HUVEC during anoxia. Anoxia-induced adhesion was prevented by addition of cycloheximide (5 micrograms/ml) to the cultures, suggesting that it depended on HUVEC protein synthesis. Enhanced adhesion was also prevented by addition of a monoclonal antibody directed against the cytokine, interleukin-1 alpha (IL-1 alpha). These findings are consistent with a role of endogenous IL-1 alpha as a mediator of the anoxia-induced adhesion of PMN to HUVEC.
Insights
Anoxia increases human polymorphonuclear leukocyte (PMN) adhesion to endothelial cells (HUVEC). This effect, mediated by interleukin-1 alpha (IL-1 alpha) and dependent on protein synthesis, is observed even after prolonged anoxia.
Area of Science:
- Cell Biology
- Immunology
- Physiology
Background:
- Endothelial cells (HUVEC) play a crucial role in regulating immune cell interactions.
- Anoxia, or low oxygen conditions, can significantly alter cellular functions and responses.
- Polymorphonuclear leukocytes (PMN) are key immune cells involved in inflammation and host defense.
Purpose of the Study:
- To investigate the effect of anoxia on the adhesion of human polymorphonuclear leukocytes (PMN) to human umbilical vein endothelial cells (HUVEC).
- To elucidate the mechanisms underlying anoxia-induced PMN-HUVEC adhesion, including the role of protein synthesis and specific cytokines.
Main Methods:
- Monolayer cultures of HUVEC were subjected to anoxic conditions for varying durations (4-8 hours).
- Adhesion of PMN to HUVEC was quantified under anoxic and normoxic conditions.
- The study utilized cycloheximide to assess the role of protein synthesis and monoclonal antibodies against interleukin-1 alpha (IL-1 alpha) to investigate cytokine involvement.
Main Results:
- Anoxia significantly increased PMN adhesion to HUVEC, with peak adhesion observed at 4-5 hours.
- Even after 8 hours of anoxia, PMN adhesion remained elevated compared to normoxic controls.
- Anoxia-induced adhesion was inhibited by cycloheximide, indicating dependence on HUVEC protein synthesis.
- The addition of an anti-IL-1 alpha antibody blocked the enhanced PMN adhesion, implicating IL-1 alpha as a mediator.
Conclusions:
- Endogenous interleukin-1 alpha (IL-1 alpha) acts as a mediator in the anoxia-induced adhesion of PMN to HUVEC.
- Anoxia triggers protein synthesis in HUVEC, leading to the expression of adhesion-promoting factors.
- These findings highlight a novel mechanism by which hypoxic conditions can enhance inflammatory cell recruitment to the endothelium.
