scholarly journals Recombinant human tissue factor pathway inhibitor exerts anticoagulant, anti-inflammatory and antibacterial effects in murine pneumococcal pneumonia

Critical Care ◽  
2009 ◽  
Vol 13 (Suppl 4) ◽  
pp. P27
Author(s):  
F Boogaard ◽  
X Brands ◽  
M Schultz ◽  
M Levi ◽  
J Roelofs ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Human Tissue ◽  
Pneumococcal Pneumonia ◽  
Tissue Factor Pathway Inhibitor ◽  
Antibacterial Effects ◽  
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 Endogenous tissue factor pathway inhibitor has a limited effect on host defence in murine pneumococcal pneumonia

2015 ◽  
Vol 114 (07) ◽  
pp. 115-122 ◽  
Author(s):  
Cornelis van ’t Veer ◽  
Joris J. T. H. Roelofs ◽  
Joost C. M. Meijers ◽  
Marcus J. Schultz ◽  
George Broze Jr ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Bacterial Growth ◽  
Pneumococcal Pneumonia ◽  
Tissue Factor Pathway Inhibitor ◽  
Community Acquired Pneumonia ◽  
Lung Pathology ◽  
Kunitz Domain ◽  
Tissue Factor Pathway ◽  
Genetic Deletion ◽  
Low Levels

SummaryStreptococcus (S.) pneumoniae is the most common causative pathogen in community-acquired pneumonia. Coagulation and inflammation interact in the host response to infection. Tissue factor pathway inhibitor (TFPI) is a natural anticoagulant protein that inhibits tissue factor (TF), the main activator of inflammation-induced coagulation. It was the objective of this study to investigate the effect of endogenous TFPI levels on coagulation, inflammation and bacterial growth during S. pneumoniae pneumonia in mice. The effect of low endogenous TFPI levels was studied by administration of a neutralising anti-TFPI antibody to wild-type mice, and by using genetically modified mice expressing low levels of TFPI, due to a genetic deletion of the first Kunitz domain of TFPI (TFPIK1(-/-)) rescued with a human TFPI transgene. Pneumonia was induced by intranasal inoculation with S. pneumoniae and samples were obtained at 6, 24 and 48 hours after infection. Anti-TFPI reduced TFPI activity by ~50 %. Homozygous lowTFPI mice and heterozygous controls had ~10 % and ~50 % of normal TFPI activity, respectively. TFPI levels did not influence bacterial growth or dissemination. Whereas lung pathology was unaffected in all groups, mice with ~10 % (but not with ~50 %) of TFPI levels displayed elevated lung cytokine and chemokine concentrations 24 hours after infection. None of the groups with low TFPI levels showed an altered procoagulant response in lungs or plasma during pneumonia. These data argue against an important role for endogenous TFPI in the antibacterial, inflammatory and procoagulant response during pneumococcal pneumonia.

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.

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