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Blood, 1 February 2001, Vol. 97, No. 3, pp. 660-668
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Changes in the biomechanical properties of neutrophils and
endothelial cells during adhesion
Qin Wang,
Eddie T. Chiang,
Mark Lim,
Jean Lai,
Rick Rogers,
Paul A. Janmey,
David Shepro, and
Claire M. Doerschuk
From the Physiology Program, Department of
Environmental Health, Harvard School of Public Health, Boston; the
Microvascular Research Laboratory, Boston University, Boston; and the
Hematology Division, Brigham and Women's Hospital, Boston, MA.
This study examined changes in the biomechanical properties
of cultured pulmonary microvascular endothelial cells (ECs) and neutrophils induced by adhesion of neutrophils to these ECs. The biomechanical properties of cells were evaluated using magnetic twisting cytometry, which measures the angular rotation of
ferromagnetic beads bound to cells through antibody ligation on
application of a specified magnetic torque. Adhesion of neutrophils to
24-hour tumor necrosis factor- (TNF-)-treated ECs, but not to
untreated ECs, induced an increase in EC stiffness within 2 minutes,
which was accompanied by an increase and a reorganization of
F-actin in ECs. A cell-permeant, phosphoinositide-binding peptide
attenuated the EC stiffening response, suggesting that intracellular
phosphoinositides are required. The stiffening response was not
inhibited by ML-7, a myosin light-chain kinase inhibitor, or BAPTA, an
intracellular Ca2+ chelator. Moreover, the phosphorylation
pattern of the regulatory myosin light chains was unaltered within 15 minutes of neutrophil adherence. These data suggested that the EC
stiffening response appeared not to be mediated by myosin
light-chain-dependent mechanisms. Concomitantly, neutrophil adhesion
to 24-hour TNF--treated ECs also induced changes in the
biomechanical properties of neutrophils compared to neutrophils bound
to untreated ECs. Taken together, these results demonstrated that
neutrophil adhesion to TNF--treated ECs induces changes in the
biomechanical properties of both cell types through actin cytoskeletal
remodeling. These changes may modulate neutrophil transmigration across
the endothelium during inflammation.
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