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-Activated Endothelium Under Flow Conditions: 4 Integrins Mediate Initial Attachment, and E-Selectin Mediates Rolling1
Departments of
*
Pulmonary Diseases and
Haematology, Academisch Ziekenhuis Utrecht, Utrecht, The Netherlands
The multistep model of leukocyte adhesion reveals that selectins
mediate rolling interactions and that integrins mediate firm adhesion
processes. In this study, the interaction between eosinophils and
TNF-
-activated HUVEC (second or third passage) was studied under
flow conditions (0.8 and 3.2 dynes/cm2). Especially the
role of 4 integrins on eosinophils and E-selectin on
HUVEC was studied. Inhibition of the integrin 4 chain on
eosinophils reduced the number of firmly adhered resting eosinophils to
TNF--stimulated endothelium by 43% whereas the percentage rolling
cells increased 2.2-fold compared with untreated control eosinophils.
Blocking of E-selectin on the endothelium reduced the number of
adherent eosinophils by only 23% and 16%. In this situation, however,
hardly any rolling adhesion was observed, and the few rolling cells
showed a low rolling velocity. Blocking both 4 integrin
on eosinophils and E-selectin on HUVEC reduced the number of adhered
eosinophils by 95%. P-selectin did not significantly participate in
eosinophil adhesion to TNF--activated HUVEC. Inhibition of both
4 integrins and ß2 integrins on
eosinophils resulted in a reduction of adhered cells by 65% and a
3-fold increase in percentage rolling cells. Taken together, these
results clearly show that resting eosinophils preferentially use
constitutively active 4 integrins
(4ß1, 4ß7)
for the first attachment to TNF--activated HUVEC. In addition,
4 integrins and E-selectin work synergistically in
eosinophil adherence to TNF--activated HUVEC. Although E-selectin is
important for eosinophil rolling under these conditions, P-selectin
plays only a minor role.
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