Immunomodulation of neutrophil–endothelial interaction by inotropes.
OBJECTIVE
To evaluate the effect of the inotropes epinephrine, dopamine and dobutamine on expression of endothelial adhesion molecules and on neutrophil adhesion to endothelial cells under dynamic conditions.


METHODS
Endothelial cells were obtained by collagenase digestion of human umbilical cord veins.Endothelial monolayers were pre-incubated with one of the chosen inotropes, with or without butoxamine, and exposed to interleukin-1. The monolayers were then incubated with fluorescencelabelled anti-human monoclonal antibodies directed against the endothelial adhesion molecules ICAM-1, E-selectin or VCAM-1. Expression of endothelial adhesion molecules was analysed by flow cytometry after pre-incubation of endothelial monolayers with one of the chosen inotropes, with or without butoxamine, and after exposure to interleukin-1. To evaluate the neutrophil adherence, the endothelium was placed on a horizontal shaker-incubator and overlayered with neutrophils. Then, non-adherent neutrophils were removed, and cells were completely dissociated. Finally, neutrophils and endothelial cells were counted by flow cytometry.


RESULTS
The expression of E-selectin on endothelium following stimulation with interleukin-1 is attenuated by the inotropes dopamine or dobutamine, but not by epinephrine. The addition of butoxamine does not modify the expression of E-selectin following stimulation with interleukin-1 and pre-incubation with one of the chosen inotropes. The decrease in neutrophil adhesion to endothelium following stimulation with interleukin-1 and addition of inotropes is antagonised by the b-blocker butoxamine.


CONCLUSIONS
In contrast to the modulation of E-selectin expression on endothelium, the effect of inotropes on neutrophil adhesion to endothelium is regulated by the expression of adhesion molecules on PMNs and mediated by the b-adrenoceptor.
Trabold
Benedikt
B
Department of Anesthesiology, University Medical Center, 93042 Regensburg, Germany.
Lunz
Dirk
D
Gruber
Michael
M
Fröhlich
Dieter
D
Graf
Bernhard
B
D002316:Cardiotonic Agents / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000276:immunology / Q000378:metabolism*; D002450:Cell Communication / Q000187:drug effects; D002478:Cells, Cultured; D004280:Dobutamine / Q000494:pharmacology; D004298:Dopamine / Q000494:pharmacology; D019040:E-Selectin / Q000276:immunology / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects* / Q000276:immunology; D004837:Epinephrine / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism; D007375:Interleukin-1 / Q000276:immunology; D009504:Neutrophils / Q000187:drug effects* / Q000502:physiology; D019010:Vascular Cell Adhesion Molecule-1 / Q000276:immunology / Q000378:metabolism
D002316:Cardiotonic Agents; D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007375:Interleukin-1; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D004280:Dobutamine; D004298:Dopamine; D004837:Epinephrine