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Physiology Program, Department of Environmental Health, Harvard School of Public Health, Boston, MA 02115
This study evaluated the changes in the biomechanical properties of
endothelial cells (ECs) induced by neutrophil adhesion and the roles of
ICAM-1 and reactive oxygen species (ROS) in modulating these changes.
Neutrophil adherence to 24-h TNF-
-activated pulmonary microvascular
ECs induced an increase in the apparent stiffness of ECs within 2 min,
measured with magnetic twisting cytometry. An anti-ICAM-1 Ab
blocked the EC stiffening response without inhibiting neutrophil
adherence. Moreover, cross-linking ICAM-1 mimicked the stiffening
response induced by neutrophils. The neutrophil-induced increase in the
apparent stiffness of ECs was inhibited with 1% DMSO (a hydroxyl
radical scavenger), allopurinol (a xanthine oxidase inhibitor), or
deferoxamine (an iron chelator), suggesting that ROS may be involved in
mediating the EC stiffening response. The cellular sources of ROS were
determined by measuring the oxidation of dichlorofluorescein.
Neutrophil adherence to TNF--activated ECs induced ROS production
only in ECs, and not in neutrophils. This ROS production in ECs was
completely prevented by the anti-ICAM-1 Ab and partially inhibited
by allopurinol. These results suggest that ICAM-1-mediated signaling
events during neutrophil adherence may activate xanthine oxidase, which
in turn mediates the ROS production in ECs that leads to stiffening.
ROS generated in ECs on neutrophil adherence appear to mediate
cytoskeletal remodeling, which may modulate subsequent inflammatory
responses.
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