Studies on the thrombogenic effects of recombinant tissue factor. In vivo versus ex vivo findings

An exposure of blood to tissue factor (TF) activates the coagulation system by the extrinsic pathway and may cause clot formation. Recombinant TF (r-TF) has been produced and subsequently reconstituted into phospholipid vesicles. The aim of these studies was to elucidate the in vitro procoagulant effects and the in vivo thrombogenicity of r-TF using a rabbit jugular vein stasis thrombosis model. The in vitro studies exhibited a clear concentration-dependent decrease in the clotting time when rabbit brain thromboplastin was replaced by r-TF in the prothrombin time assay. The in vivo studies revealed a dose-dependent thrombogenicity between 1.6 ng/kg and 50 ng/kg. Electron microscope scanning of the surface of representative clots revealed fibrin-rich structures of heterogeneous density. In comparison, thrombi obtained when FEIBA was utilized as the thrombogenic agent were more homogeneous. The injection of r-TF caused a slight transient drop in blood pressure with little or no effects on the pulse rate, complete blood count (CBC) profile, clotting and amidolytic assays when compared to sham control animals. In contrast, the whole blood clotting parameters (activated clotting time and thrombelastograph) were prolonged dose-dependently after r-TF injection. The antithrombotic activity of heparin was assessed in this model and compared to the antithrombotic activity when FEIBA is used as the thrombogenic agent. The apparent ED50 of heparin was found to be 4 times higher in the r-TF system. In control studies, no thrombogenic effects were observed by the phospholipid vesicles alone nor by r-TF not embedded in phospholipid vesicles. These data demonstrate that lipidated r-TF is a potent thrombogenic challenge that activates the hemostatic system by the extrinsic pathway.