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Updated: Jul 13, 2026
Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions
Published on: August 8, 2012
Interactions of neutrophils and endothelial cells under low flow conditions in vitro
N Yoshida1, T Yoshikawa, Y Nakamura
1First Department of Internal Medicine, Kyoto Prafectural University of Medicine, Japan.
Abstract:
The interactions of polymorphonuclear leukocytes (PMN) and endothelial cells are modulated by adhesion molecules, inflammatory cytokines, and shear stress. We investigated the changes in PMN-endothelial cell interactions induced by interleukin (IL)-1 beta under low flow conditions. PMN were isolated from the venous blood of healthy adults, and endothelial cells were obtained from human umbilical veins. The number of PMN that adhered to the endothelial cells monolayer that was treated with IL-1 increased significantly at shear stresses from .5 to 4.0 dyn/cm2 as compared with untreated endothelial cells. Anti-intercellular adhesion molecule (ICAM)-1 monoclonal antibody (mAb), anti-E-selectin mAb, and anti-CD18 mAb each significantly inhibited the increase in PMN adherence induced by IL-1 at a low shear stress (1.0 dyn/cm2). Anti-CD18 mAb significantly reduced the number of PMN that migrated through the endothelial monolayer by blocking the adherence of PMN to the luminal surface of the endothelial cells, as well as their transendothelial migration. In contrast, anti-ICAM-1 and anti-E-selectin mAb each reduced the number of PMN that migrated by reducing the number of PMN that adhered to the luminal surface without significantly influencing the percent of the adherent PMN that had migrated. Although anti-L-selectin mAb reduced the adherence and migration of PMN, these effects were not statistically significant. These results indicated that under low flow conditions, as well as in the nonflow state, PMN-endothelial cell interactions were elicited via CD11/CD18 and ICAM-1 without the involvement of selectins.
Insights
Interleukin-1 beta increases polymorphonuclear leukocyte (PMN) adherence to endothelial cells under low flow. CD11/CD18 and ICAM-1 mediate these interactions, not selectins.
Area of Science:
- Immunology
- Cell Biology
- Biomedical Engineering
Background:
- Polymorphonuclear leukocyte (PMN) and endothelial cell interactions are crucial in inflammation.
- These interactions are influenced by shear stress, cytokines, and adhesion molecules.
Purpose of the Study:
- To investigate the effect of interleukin-1 beta (IL-1 beta) on PMN-endothelial cell interactions under low flow conditions.
- To identify the specific adhesion molecules involved in IL-1 beta-induced PMN recruitment.
Main Methods:
- Human umbilical vein endothelial cells and PMN from healthy adults were used.
- PMN adherence and transmigration assays were performed under varying shear stresses (0.5-4.0 dyn/cm2) with and without IL-1 beta.
- Monoclonal antibodies against ICAM-1, E-selectin, CD18, and L-selectin were used to block specific adhesion molecules.
Main Results:
- IL-1 beta significantly increased PMN adherence to endothelial cells at low shear stress.
- Anti-ICAM-1, anti-E-selectin, and anti-CD18 antibodies inhibited IL-1 beta-induced PMN adherence.
- Anti-CD18 antibody significantly reduced both PMN adherence and transmigration.
- Anti-ICAM-1 and anti-E-selectin antibodies reduced transmigration by decreasing adherence, but did not affect the transmigration percentage of adherent PMN.
- Anti-L-selectin antibody showed non-significant effects on PMN adherence and migration.
Conclusions:
- Under low flow, IL-1 beta enhances PMN-endothelial cell interactions primarily through CD11/CD18 and ICAM-1.
- Selectins do not play a significant role in IL-1 beta-induced PMN recruitment under these conditions.
- These findings elucidate key molecular mechanisms in inflammatory cell recruitment at low shear stress.
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