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Activated factor X and thrombin formation triggered by tissue factor on
endothelial cell matrix in a flow model: effect of the tissue factor
pathway inhibitor
C van 't Veer, TM Hackeng, C Delahaye, JJ Sixma and BN Bouma
Department of Hematology, University Hospital Utrecht, The Netherlands.
The procoagulant subcellular matrix of stimulated endothelial cells that
contains tissue factor (TF) was used to investigate the mechanism by which
TF pathway inhibitor (TFPI) inhibits thrombin formation initiated by
TF/factor VIIa (FVIIa) under flow conditions. Purified coagulation factors
VII, X, and V and prothrombin were perfused at a wall shear rate of 100 s-1
through a flow chamber containing a coverslip covered with matrix of
cultured human umbilical vein endothelial cells. This resulted in a TF- and
FVII-dependent FXa and thrombin generation as measured in the effluent at
the outlet of the system. Inhibition of this TF/FVIIa-triggered thrombin
formation by TFPI purified from plasma was dependent on the amount of TF
present on the endothelial cell matrix. The rate of prothrombinase assembly
and steady-state levels of thrombin formation were decreased by TFPI.
Because persistent albeit decreased steady-state levels of thrombin
formation occurred in the presence of TFPI, we conclude that plasma- TFPI
does not inhibit FXa present in the prothrombinase complex. The addition of
FIX and FVIII to perfusates containing FVII and FX increased the FXa
generation on endothelial matrices, and counteracted the inhibition of
thrombin formation on endothelial cell matrices by TFPI. Our data provide
further evidence for the hypothesis that the rapid inactivation of TF/FVIIa
by TFPI in combination with the absence of either FVIII or FIX causes the
bleeding tendency of patients with hemophilia A or B.
Volume 84,
Issue 4,
pp. 1132-1142,
08/15/1994
Copyright © 1994 by The American Society of Hematology
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