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Tumor necrosis factor-induced endothelial tissue factor is associated with
subendothelial matrix vesicles but is not expressed on the apical surface
J Ryan, J Brett, P Tijburg, RR Bach, W Kisiel and D Stern
Department of Physiology and Cellular Biophysics, Columbia University,
College of Physicians and Surgeons, New York, NY 10032.
Cultured endothelial cells can be induced by tumor necrosis
factor/cachectin (TNF) and other cytokines to synthesize the procoagulant
cofactor tissue factor (TF). Intact monolayers of TNF- treated endothelial
cells showed only minimal TF activity. In contrast, after permeabilization
of these monolayers with detergent (saponin, 0.02%), there was
approximately 10- to 20-fold increase in TF-mediated, factor VIIa-dependent
factor Xa formation. Extracellular matrix derived from TNF-treated
endothelium, prepared after removing the cells by hypotonic lysis or
ammonium hydroxide (0.1 N), also had similarly enhanced TF activity.
Incubation with a blocking monoclonal antibody to TF inhibited the
procoagulant activity of both TNF-stimulated endothelial cells, whether
they were intact or permeabilized, and of their matrices. However, when the
apical cell surface was pretreated with anti-TF antibody, washed, and then
cells were lysed with water or permeabilized with saponin, similar
augmentation of TF activity was still observed, suggesting the presence of
a pool of TF to which the antibody did not initially gain access.
Consistent with this concept, the presence of TF in the matrix of
TNF-treated endothelial cells was shown by immunoblotting and morphologic
studies; cultured endothelial monolayers and the native endothelium of
aortic segments after exposure to TNF showed TF in extracellular matrix,
associated with vesicles. In contrast, TF was virtually undetectable on the
apical endothelial surface. Taken together, these findings suggest that
endothelial TF can be present in a cryptic pool that only gains access to
the blood after alteration in the integrity of the endothelial monolayer.
Volume 80,
Issue 4,
pp. 966-974,
08/15/1992
Copyright © 1992 by The American Society of Hematology
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