Nebulized Recombinant Human Tissue Factor Pathway Inhibitor Attenuates Coagulation and Exerts Modest Anti-inflammatory Effects in Rat Models of Lung Injury

2017 ◽  
Vol 30 (2) ◽  
pp. 91-99 ◽  
Author(s):  
Florry E. van den Boogaard ◽  
Jorrit J. Hofstra ◽  
Xanthe Brands ◽  
Marcel M. Levi ◽  
Joris J.T.H. Roelofs ◽  
...  
Keyword(s):  
Lung Injury ◽  
Tissue Factor ◽  
Human Tissue ◽  
Tissue Factor Pathway Inhibitor ◽  
Rat Models ◽  
Tissue Factor Pathway ◽  
Anti Inflammatory

scholarly journals Thromboresistance of Balloon-Injured Porcine Carotid Arteries After Local Gene Transfer of Human Tissue Factor Pathway Inhibitor

Circulation ◽  
2000 ◽  
Vol 101 (3) ◽  
pp. 289-295 ◽  
Author(s):  
Pierre Zoldhelyi ◽  
Janice McNatt ◽  
Harnath S. Shelat ◽  
Yasutaka Yamamoto ◽  
Zhi-Qiang Chen ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Tissue Factor ◽  
Human Tissue ◽  
Tissue Factor Pathway Inhibitor ◽  
Carotid Arteries ◽  
Tissue Factor Pathway

scholarly journals Enhanced Antifibrinolytic Efficacy of a Plasmin-Specific Kunitz-Inhibitor (60-Residue Y11T/L17R with C-Terminal IEK) of Human Tissue Factor Pathway Inhibitor Type-2 Domain1

2020 ◽  
Vol 9 (11) ◽  
pp. 3684
Author(s):  
Kanagasabai Vadivel ◽  
Anne K. Zaiss ◽  
Yogesh Kumar ◽  
Frank M. Fabian ◽  
Ayman E. A. Ismail ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Human Tissue ◽  
Active Sites ◽  
Tissue Factor Pathway Inhibitor ◽  
Clot Lysis ◽  
Dependent Manner ◽  
Kunitz Inhibitor ◽  
Tissue Factor Pathway ◽  
Inhibitor Type

Current antifibrinolytic agents reduce blood loss by inhibiting plasmin active sites (e.g., aprotinin) or by preventing plasminogen/tissue plasminogen activator (tPA) binding to fibrin clots (e.g., ε-aminocaproic acid and tranexamic acid); however, they have adverse side effects. Here, we expressed 60-residue (NH2NAE…IEKCOOH) Kunitz domain1 (KD1) mutants of human tissue factor pathway inhibitor type-2 that inhibit plasmin as well as plasminogen activation. A single (KD1-L17R-KCOOH) and a double mutant (KD1-Y11T/L17R- KCOOH) were expressed in Escherichia coli as His-tagged constructs, each with enterokinase cleavage sites. KD1-Y11T/L17R-KCOOH was also expressed in Pichia pastoris. KD1-Y11T/L17R-KCOOH inhibited plasmin comparably to aprotinin and bound to the kringle domains of plasminogen/plasmin and tPA with Kd of ~50 nM and ~35 nM, respectively. Importantly, compared to aprotinin, KD1-L17R-KCOOH and KD1-Y11T/L17R-KCOOH did not inhibit kallikrein. Moreover, the antifibrinolytic potential of KD1-Y11T/L17R-KCOOH was better than that of KD1-L17R-KCOOH and similar to that of aprotinin in plasma clot-lysis assays. In thromboelastography experiments, KD1-Y11T/L17R-KCOOH was shown to inhibit fibrinolysis in a dose dependent manner and was comparable to aprotinin at a higher concentration. Further, KD1-Y11T/L17R-KCOOH did not induce cytotoxicity in primary human endothelial cells or fibroblasts. We conclude that KD1-Y11T/L17R-KCOOH is comparable to aprotinin, the most potent known inhibitor of plasmin and can be produced in large amounts using Pichia.

scholarly journals Anti-inflammatory effects of long-lasting locally-delivered human recombinant tissue factor pathway inhibitor after balloon angioplasty

2002 ◽  
Vol 97 (3) ◽  
pp. 198-205 ◽  
Author(s):  
Yoichi Nakamura ◽  
Kazufumi Nakamura ◽  
Keiko Ohta ◽  
Hiromi Matsubara ◽  
Chikao Yutani ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Balloon Angioplasty ◽  
Tissue Factor Pathway Inhibitor ◽  
Tissue Factor Pathway ◽  
Anti Inflammatory

Human Tissue Factor Pathway Inhibitor-2 Induces Caspase-Mediated Apoptosis in a Human Fibrosarcoma Cell Line (HT-1080).

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4165-4165
Author(s):  
Prakasha Kempaiah ◽  
Walter Kisiel
Keyword(s):  
Tissue Factor ◽  
Human Tissue ◽  
Tissue Factor Pathway Inhibitor ◽  
Serine Proteinase ◽  
Annexin V ◽  
Polymyxin B ◽  
Bax Protein ◽  
Tissue Factor Pathway ◽  
Kunitz Type

Abstract Human tissue factor pathway inhibitor-2 (TFPI-2) is a 32 kDa extracellular matrix-associated Kunitz-type serine proteinase inhibitor that regulates the plasmin and trypsin- mediated activation of zymogen matrix metalloproteinases and growth factors essential for tumor growth, invasion, and metastasis. We previously demonstrated that HT-1080 human fibrosarcoma cells do not express TFPI-2, but restoration of TFPI-2 expression in these cells by stable transfection with the human TFPI-2 cDNA markedly inhibited their growth and metastasis in-vivo. In this study, 1 μM concentrations of either recombinant human TFPI-2 or its mutated first Kunitz-type domain (R24K KD1), were offered to HT-1080 cells, incubated for 48 h, and the degree of apoptosis assessed by nuclear fragmentation, ethidium bromide/acridine orange (EB/AO) staining, fluorescence-activated cell sorting (FACS) and immunoblotting. Agarose gel electrophoresis of DNA from HT-1080 cells treated with either TFPI-2 or R24K KD1 for 48h indicated DNA fragmentation with a ladder pattern typically associated with apoptosis. EB/AO staining of TFPI-2- and R24K KD1- treated cells revealed that 40–70% of the cells were apoptotic in relation to vehicle-treated cells as judged by fluoresence microscopy. Co-administration of TFPI-2 with polymyxin B produced similar results, suggesting that apoptosis was not endotoxin-dependent. Consistent with our earlier studies showing its enhanced inhibitory activity relative to TFPI-2, R24K KD1 was able to induce apoptosis in 68% of HT-1080 cells after 48h of treatment compared to 39% for the parent TFPI-2 at an equivalent concentration. Moreover, HT-1080 cells treated with a KD1 preparation lacking the reactive site arginine residue (R24Q KD1) produced only an 18% apoptosis rate, thereby linking the observed apoptosis with serine proteinase inhibition. FACS analysis, using propidium iodide and annexin-V labeling, revealed similar apoptotic rates to that seen by EB/AO staining assays. In addition, immunoblotting experiments of vehicle and TFPI-2-treated cells indicated increased caspase-3 activation in TFPI-2-treated cells, thus providing evidence that apoptosis is caspase-mediated. We also observed up-regulation of the proapoptotic Bax protein by immunoblotting following treatment of HT-1080 cells with either TFPI-2 or R24K KD1. Taken together, our results demonstrate that treatment of HT-1080 cells with exogenous TFPI-2 or R24K KD1 activates caspase-mediated, proapoptotic signaling pathways and induces apoptosis. In addition, our data provides suggestive evidence that peritumor application of either TFPI-2 or R24K KD1 may facilitate tumor apoptosis in-vivo.

Human Tissue Factor Pathway Inhibitor (TFPI) Gene: Complete Genomic Structure and Localization on the Genetic Map of Chromosome 2q

Genomics ◽  
1993 ◽  
Vol 17 (2) ◽  
pp. 423-428 ◽  
Author(s):  
Kei-ichi Enjyoji ◽  
Mitsuru Emi ◽  
Tsunehiro Mukai ◽  
Motohiro Imada ◽  
Mark L. Leppert ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Genetic Map ◽  
Human Tissue ◽  
Tissue Factor Pathway Inhibitor ◽  
Genomic Structure ◽  
Tissue Factor Pathway ◽  
Complete Genomic
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