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Migration of primed human eosinophils across cytokine-activated endothelial
cell monolayers
R Moser, J Fehr, L Olgiati and PL Bruijnzeel
Department of Medicine, University Hospital, Zurich, Switzerland.
Eosinophils are known to adhere to cytokine-activated endothelium. Whereas
transendothelial migration for neutrophils is an inevitable consequence of
this endothelial-dependent adherence, this has not yet been shown for
eosinophils. By means of human umbilical vein endothelial cells (HUVE)
grown to confluence on microporous filters as an in vitro model of
leukocytic migration across postcapillary venules, we have characterized
the conditions leading to endothelium-driven transmigration of blood
eosinophils from normals and from patients with allergic asthma. Freshly
isolated eosinophils from nonallergic donors adhered to interleukin-1
(IL-1) and tumor necrosis factor-activated HUVE, but did not penetrate
these monolayers. In contrast, eosinophils from allergic asthma patients
showed an increased adherence and transmigration capacity. This increased
functional competence was not caused by a difference in density phenotype,
because the eosinophils from both groups showed a comparable density
distribution over discontinuous Percoll gradients. Moreover, no difference
existed within one group among eosinophils harvested from the Percoll
density bands 1.080, 1.085, and 1.090 g/mL in terms of transendothelial
migration. In vitro cultivation of freshly isolated eosinophils from
nonallergic individuals in the presence of granulocyte-macrophage colony-
stimulating factor (GM-CSF) and IL-3 induced a stepwise decrease of the
density distribution over such gradients. In contrast, eosinophils from
patients with allergic asthma directly shifted to a final density of 1.075
g/mL within 24 hours of culture. Notwithstanding the kinetics of density
changes, eosinophils from nonallergic donors already expressed the capacity
to transmigrate IL-1-activated HUVE monolayers 20 hours after cultivation
with different combinations of GM-CSF, IL-3, and IL- 5. Inhibition studies
with monoclonal antibodies showed that endothelium-driven transmigration of
eosinophils predominantly implicates CD11/CD18 structures on the eosinophil
surface, whereas no significant inhibition was found with the anti-VLA-4
monoclonal antibody HP2/1. From cytofluorometric studies, we conclude that
spontaneous transmigration of eosinophils from allergic asthma patients is
not accompanied by quantitative upregulation of these antigens. Taken
together, these results allow the conclusion that blood eosinophils from
allergic asthma patients have undergone in vivo priming, mimicked in vitro
by cytokines such as GM-CSF, IL-3, and IL-5, leading to induction of the
capacity to migrate across cytokine- activated HUVE monolayers.
Volume 79,
Issue 11,
pp. 2937-2945,
06/01/1992
Copyright © 1992 by The American Society of Hematology
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