scholarly journals Retinoic acid reduces induction of monocyte tissue factor and tissue factor/factor VIIa-dependent arterial thrombus formation

Blood ◽  
1995 ◽  
Vol 86 (1) ◽  
pp. 212-218 ◽  
Author(s):  
RM Barstad ◽  
MJ Hamers ◽  
RW Stephens ◽  
KS Sakariassen
Keyword(s):  
Retinoic Acid ◽  
Tissue Factor ◽  
Human Peripheral Blood ◽  
Thrombus Formation ◽  
Antigen Expression ◽  
Fibrin Deposition ◽  
Thrombotic Risk ◽  
Plaque Disruption ◽  
Platelet Thrombus ◽  
Arterial Grafting

Agents that downregulate the induction of monocyte/macrophage tissue factor (TF) activity may attenuate the thrombotic risk associated with mechanical restoration of vessel patency or artificial arterial grafting. In such events, procoagulant macrophages in the atherosclerotic plaque and procoagulant monocytes adherent to artificial materials may be exposed to the blood stream. Ishii et al (Blood 80:2556, 1992) reported that induction of endothelial TF is downregulated by all-trans retinoic acid (ATRA), and Conese et al (Thromb Haemost 66:662, 1991) reported that retinoids downregulate monocyte procoagulant activity (PCA). These findings led us to investigate the effect of ATRA on monocyte TF expression, and to study the effect of ATRA on monocyte-induced thrombus formation in a model system of human arterial thrombogenesis. Induction of PCA in human peripheral blood monocytes by 0.5 microgram/mL lipopolysaccharide (LPS) was dose dependently reduced by ATRA, reaching a reduction of 56% at 10(-5) mol/L ATRA (P < .0001). A 38% reduction (P < .0007) in LPS- induced TF antigen expression was observed at an ATRA concentration of 10(-6) mol/L. Adherence of monocytes to plastic cover slips (Thermanox, Miles Laboratories, Naperville, IL) also triggered induction of cellular PCA, which was inhibited by more than 80% by an anti-TF monoclonal antibody (MoAb) (P < .002). Inclusion of ATRA (10(-6) mol/L) reduced this PCA by 40% (P < .03), and the TF antigen expression by 30% (P < .0001). Exposure of Thermanox adherent monocytes to flowing nonanticoagulated human blood in a parallel-plate perfusion chamber device at an arterial wall shear rate of 650 s-1 elicited significant fibrin deposition and platelet thrombus formation. Partial interruption of this thrombus formation was achieved by 10(-6) mol/L ATRA, which reduced the fibrin deposition by 80% (P < .02) and platelet thrombus formation by 50% (P < .05). In comparison, incubation of adherent monocytes with the anti-TF MoAb before the blood exposure, reduced the fibrin deposition by 83% (P < .02) and platelet thrombus volume by 75% (P < .0008). Thus, ATRA is an effective down-regulator of monocyte TF- PCA, and may reduce thrombotic complications at sites of plaque rupture, at plaque disruption after percutaneous transluminal angioplasty procedures, or on surfaces introduced by artificial arterial grafting.

scholarly journals Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin

2003 ◽  
Vol 197 (11) ◽  
pp. 1585-1598 ◽  
Author(s):  
Shahrokh Falati ◽  
Qingde Liu ◽  
Peter Gross ◽  
Glenn Merrill-Skoloff ◽  
Janet Chou ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Blood Coagulation ◽  
Thrombus Formation ◽  
Recipient Mouse ◽  
Wild Type ◽  
Activated Platelets ◽  
Injury Model ◽  
Platelet Thrombus ◽  
Flowing Blood

Using a laser-induced endothelial injury model, we examined thrombus formation in the microcirculation of wild-type and genetically altered mice by real-time in vivo microscopy to analyze this complex physiologic process in a system that includes the vessel wall, the presence of flowing blood, and the absence of anticoagulants. We observe P-selectin expression, tissue factor accumulation, and fibrin generation after platelet localization in the developing thrombus in arterioles of wild-type mice. However, mice lacking P-selectin glycoprotein ligand 1 (PSGL-1) or P-selectin, or wild-type mice infused with blocking P-selectin antibodies, developed platelet thrombi containing minimal tissue factor and fibrin. To explore the delivery of tissue factor into a developing thrombus, we identified monocyte-derived microparticles in human platelet–poor plasma that express tissue factor, PSGL-1, and CD14. Fluorescently labeled mouse microparticles infused into a recipient mouse localized within the developing thrombus, indicating that one pathway for the initiation of blood coagulation in vivo involves the accumulation of tissue factor– and PSGL-1–containing microparticles in the platelet thrombus expressing P-selectin. These monocyte-derived microparticles bind to activated platelets in an interaction mediated by platelet P-selectin and microparticle PSGL-1. We propose that PSGL-1 plays a role in blood coagulation in addition to its known role in leukocyte trafficking.

Protein Disulfide Isomerase Is Required for Fibrin Generation and Platelet Thrombus Formation In Vivo.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 292-292 ◽  
Author(s):  
Jaehyung Cho ◽  
Barbara C. Furie ◽  
Shaun R. Coughlin ◽  
Bruce Furie
Keyword(s):  
Tissue Factor ◽  
Platelet Activation ◽  
Thrombus Formation ◽  
Null Mouse ◽  
Fibrin Formation ◽  
Platelet Thrombus ◽  
Wall Injury ◽  
Protein Disulfide ◽  
Dose Dependent

Abstract Thiol isomerases catalyze disulfide oxidation, reduction and isomerization, playing an important role during protein synthesis. Recent studies suggest a role for protein disulfide isomerase (PDI), a prototype of the thiol isomerase family, in platelet function and regulation of tissue factor activity (Essex and Li. Curr Drug Targets. 2006; Chen and Hogg. J Thromb Haemost. 2006). To determine the role of intravascular PDI during thrombus formation, PDI expression, platelet accumulation, and fibrin generation were monitored following laser-induced arteriolar injury in the mouse cremaster muscle by intravital fluorescence microscopy. PDI antigen exhibited a time-dependent increase in the developing thrombus after vessel wall injury and remained associated with the thrombus. Infusion of bacitracin, a non-specific inhibitor of thiol isomerases, into the circulation inhibited platelet thrombus formation and fibrin generation in a dose-dependent manner. Infusion of a function-blocking monoclonal antibody to PDI (RL90) into the circulation of a wild type mouse also resulted in dose-dependent inhibition of platelet accumulation and fibrin generation. To determine whether PDI inhibits fibrin formation by blocking tissue factor activation, or by preventing platelet activation and the development of the membrane surface that is required for assembly of the tenase and the prothrombinase complex in vivo, we explored fibrin formation in mice lacking protease-activated receptor-4 (Par4). Although there is no stable accumulation of platelets and no platelet activation, fibrin formation is normal in the Par4 null mouse (Vandendries et al, Proc Natl Acad Sci USA. 2007), suggesting that fibrin generation in the laser-induced vessel injury model is independent of platelet activation. Infusion of the function-blocking anti-PDI antibody (RL90) into the circulation of a Par4 null mouse prior to vessel wall injury inhibited fibrin generation. These results indicate that PDI is required to generate tissue factor in a form that leads to thrombin generation and fibrin formation during thrombus development and is required for thrombus formation.

scholarly journals Procoagulant Human Monocytes Mediate Tissue Factor/Factor VIIa–Dependent Platelet-Thrombus Formation When Exposed to Flowing Nonanticoagulated Human Blood

1995 ◽  
Vol 15 (1) ◽  
pp. 11-16 ◽  
Author(s):  
R. Marius Barstad ◽  
Maria J. A. G. Hamers ◽  
Peter Kierulf ◽  
Åse-Britt Westvik ◽  
Kjell S. Sakariassen
Keyword(s):  
Tissue Factor ◽  
Human Blood ◽  
Factor Viia ◽  
Thrombus Formation ◽  
Human Monocytes ◽  
Platelet Thrombus

Factor XI Deficient Mice Have Reduced Platelet Accumulation and Fibrin Deposition after Laser Injury.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 218-218
Author(s):  
T. Regan Baird ◽  
David Gailani ◽  
Bruce Furie ◽  
Barbara C. Furie
Keyword(s):  
Tissue Factor ◽  
Factor Viia ◽  
Thrombus Formation ◽  
Factor Xa ◽  
Fibrin Deposition ◽  
Wild Type ◽  
Factor Xi ◽  
Factor Xia ◽  
Tissue Factor Pathway ◽  
Platelet Accumulation

Abstract Tissue factor exposure at sites of vascular injury results in the generation of factor Xa and thrombin. A current model of blood coagulation suggests that the amount of thrombin generated through this pathway is limited by the inhibition of the factor VIIa-tissue factor complex by tissue factor pathway inhibitor in the presence of factor Xa. The initial thrombin activates a number of hemostatic proteins including factor XI. Factor XIa then activates factor IX leading to generation of the tenase complex to maintain the thrombin flux. While in vitro studies support this hypothesis the importance of factor XI for thrombus formation in vivo remains unclear. We have examined thrombus formation upon laser injury to the arterioles (30–50 μm diameter) of the cremaster muscle in living mice lacking factor XI using digital multi-channel fluorescence intravital microscopy. Platelets were labeled with Alexa 488 conjugated murine CD41 Fab fragments by systemic infusion of the antibody. Maximum platelet accumulation in factor XI null mice (median of 35 thrombi in 4 mice) is only 25% of that of wild type mice (median of 40 thrombi in 4 mice) after injury (p<0.03). The time course of platelet accumulation is similar between both genotypes. Maximum platelet accumulation occurs in approximately 90 seconds (p<0.2). Fibrin deposition was observed simultaneously using an Alexa 660 conjugated anti-fibrin antibody that does not recognize fibrinogen. Maximum fibrin deposition in factor XI null mice is 50% that of wild type mice (p<0.001) and the rate of fibrin generation is slower in factor XI null mice. However, the time to achieve half maximal fibrin deposition is approximately the same in factor XI null mice (77 sec) compared to wild type mice (63.5 sec, p<0.09). These data suggest that the primary difference in response to laser induced injury between the factor XI null mice and wild type mice is the level of thrombin generated and supports the hypothesis that factor XI participates in maintaining thrombin flux after inhibition of the factor VII-tissue factor. The model above postulates a single source of tissue factor, the vessel wall, and further, that the tissue factor-factor VIIa complex formed from the exposed tissue factor is rapidly inactivated by tissue factor pathway inhibitor after the appearance of the initial factor Xa formed. In addition it has been suggested that a rapidly growing thrombus blocks access to vascular wall tissue factor. However we have recently observed that there is a P-selectin and P-selectin glycoprotein ligand 1 dependent pathway of blood coagulation that recruits blood borne tissue factor into a growing thrombus at sites of laser-induced vessel injury. Both vessel wall and blood borne tissue factor are required for normal thrombus formation. Our data suggest that although tissue factor is continuously recruited to the growing thrombus, factor XIa plays a significant role in thrombin generation.

Iohexol, platelet activation and thrombosis

1998 ◽  
Vol 39 (4) ◽  
pp. 355-361 ◽  
Author(s):  
K. S. Sakariassen ◽  
M. Buchmann ◽  
M. J. A. G. Hamers ◽  
H. Stormorken
Keyword(s):  
Tissue Factor ◽  
Platelet Activation ◽  
Thrombus Formation ◽  
Initial Study ◽  
Human Model ◽  
Thrombotic Risk ◽  
Arterial Thrombus ◽  
Radiographic Contrast ◽  
Radiographic Contrast Media

Background: There is a dispute about the potential effects of radiographic contrast media (CM) on thrombogenesis. The nonionic CM iohexol triggers platelet β-thromboglobulin (β-TG) secretion, and thus may activate the platelets and promote thrombosis. We addressed this topic in a study employing a human model of arterial thrombus formation in the presence of aspirin and heparin. This was a follow-up to our initial study (on thrombus formation in native blood) which did not include antithrombotic drugs. The nonionic CM iohexol (monomer) and iodixanol (dimer) and the ionic CM ioxaglate (dimer) were compared. Methods and Results: Thrombus formation was triggered by a surface rich in either collagen or tissue factor, positioned in a parallel-plate perfusion chamber device at an arterial wall shear rate of 2600 s−1. Blood from healthy volunteers, following ingestion of 1 g aspirin, was mixed with 40 vol% CM and 2.0 IU/ml heparin and passed over the surfaces. Thrombus formation in the presence of either CM showed no difference, despite the fact that iohexol triggered a pronounced platelet β-TG secretion; iodixanol or ioxaglate were virtually inert. Conclusion: There was no association between iohexol-induced β-TG secretion and thrombus formation on collagen (platelet-driven) or on tissue factor (thrombin-driven) in the presence of a standard antithrombotic regimen of aspirin and heparin as used in the clinic. The notion of a thrombotic risk due to platelet activation by iohexol was thus not substantiated by this study.

scholarly journals Abelacimab Reduces Thrombus Propagation in a Baboon Model of Vascular Graft Thrombosis

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 1027-1027
Author(s):  
Michael Wallisch ◽  
Yasser Khder ◽  
Monica T. Hinds ◽  
Erik I Tucker ◽  
Dan Bloomfield ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Tissue Factor ◽  
Thrombus Formation ◽  
Therapeutic Benefit ◽  
Clot Formation ◽  
Fibrin Deposition ◽  
Publicly Traded ◽  
Graft Thrombosis ◽  
Av Shunt ◽  
Collagen Tissue

Abstract Background: Factor XI (FXI) inhibition demonstrated strong efficacy in preventing thrombus formation in preclinical and clinical models of arterial and venous thrombosis. However, the effect of FXI inhibition in halting the progression of a formed clot remains largely unknown. Aims: This study aims to test whether abelacimab, a dual-acting FXI and activated FXI (FXIa) monoclonal antibody, is effective in halting clot formation and downstream growth when administered before or during active clot formation in an established baboon femoral arterio-venous (AV) shunt model. Methods: Three baboons had a chronic femoral AV shunt put in place; platelet and fibrin deposition inside and distal to collagen- or collagen + tissue factor (TF)-coated vascular grafts were measured at baseline (control), in a therapeutic setting, where abelacimab (1 mg/kg, intravenously) was administered 30 minutes after thrombus initiation, and in a preventative setting within the first 48 h and 1 week (144 - 216 h) post-administration. Pharmacodynamic effect was measured by activated partial thromboplastin time (aPTT). Results: Consistent with its half-life of 20 to 30 days, single iv administration of abelacimab at a dose of 1 mg/kg resulted in long-lasting (&gt; 4-week) aPTT prolongation (&gt; 2-fold). Administration of abelacimab 30 minutes after initiation of thrombosis using grafts coated either with collagen or with collagen + TF quickly halted downstream propagation of platelet and fibrin deposition compared to control. Further, downstream propagation of platelet and fibrin deposition was markedly reduced when clotting was induced by collagen, or collagen + tissue factor after abelacimab administration. Conclusions: These data suggest that abelacimab, a dual-acting anti-FXI/FXIa monoclonal antibody with a single long-lasting iv injection has the potential to slow down the growth and reduce the size of thrombi when admistered before or after clot induction. Data indicate a potential for therapeutic benefit of targeting FXI both in therapeutic and preventive settings. Sponsored by: Anthos Therapeutics Inc., 55 Cambridge Parkway, Suite 103, Cambridge, MA 02142 Figure 1 Figure 1. Disclosures Wallisch: Aronora Inc,: Current Employment. Khder: Anthos Therapeutics: Consultancy; Novartis: Current equity holder in publicly-traded company, Other: Retiree. Bloomfield: Anthos Therapeutics: Current Employment. Gruber: Aronora Inc.: Current Employment, Current equity holder in publicly-traded company; Oregon Health and Science University: Current Employment.

scholarly journals Human Coronary Thrombus Formation Is Associated With Degree of Plaque Disruption and Expression of Tissue Factor and Hexokinase II

2015 ◽  
Vol 79 (11) ◽  
pp. 2430-2438 ◽  
Author(s):  
Nozomi Okuyama ◽  
Shuntaro Matsuda ◽  
Atsushi Yamashita ◽  
Sayaka Moriguchi-Goto ◽  
Naoki Sameshima ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Thrombus Formation ◽  
Hexokinase Ii ◽  
Plaque Disruption ◽  
Coronary Thrombus

scholarly journals Animal Models of Arterial and Venous Thrombosis

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. SCI-2-SCI-2
Author(s):  
Bruce Furie ◽  
Barbara C. Furie ◽  
Robert Flaumenhaft ◽  
Jeffrey I. Zwicker
Keyword(s):  
Animal Models ◽  
Tissue Factor ◽  
Intravital Microscopy ◽  
Vessel Wall ◽  
High Speed ◽  
Thrombus Formation ◽  
Critical Role ◽  
Receptor Activation ◽  
Platelet Receptor ◽  
Platelet Thrombus

The intersection of the availability of genetically altered mice, new technologies for intravital microscopy and high speed computing systems has led to the use of animal models to build on the concepts that have emerged from in vitro studies of the molecular and cellular biology of hemostasis and thrombosis. In an effort to improve the understanding of the etiology and pathogenesis of thrombosis, thrombus formation has been initiated in experimental systems via mechanical disruption, laser-induced, photochemical-induced and ferric chloride-induced injury to the vessel wall, among others. None of these methods are physiologic, and as such, remain models that require extrapolation from a living animal – a mouse – to human biology. We have focused on laser-induced injury of the arteriole vessel wall in the cremasteric muscle of the mouse. Using high speed digital imaging of fluorescently labeled components and real time intravital microscopy, our group has been able to demonstrate that platelet accumulation and fibrin generation during thrombus formation occur simultaneously, that tissue factor and collagen are independent initiators of platelet activation, and that monocyte-derived microparticles deliver tissue factor to the site of thrombus development. Perhaps the most important and unanticipated observation has been that thiol isomerases, thought only to be involved in protein biosynthesis via the formation of disulfide bonds in the endoplasmic reticulum, play a critical role in thrombus formation. Protein disulfide isomerase (PDI), ERp5 and ERp57 are among the vascular thiol isomerases that are known to be important for the initiation of thrombus formation. Laser-induced thrombosis in mice is associated with PDI, ERp5 and ERp57 secretion by platelets and endothelium. Inhibition of these thiol isomerases blocks platelet thrombus formation and fibrin generation. The integrins αIIbβ3 and αVβ3 play a key role in this process, binding directly with the secreted thiol isomerases and capturing them in the vicinity of vessel wall injury. These enzymes are required for the initiation of platelet thrombus formation and fibrin generation, but the mechanism by which they function remains to be elucidated. At present, it would appear that there is an electron transfer pathway involving these enzymes that regulates the initiation of thrombus formation. The mechanism by which PDI participates in thrombus generation is being evaluated by using trapping mutant forms to identify substrates of PDI that participate in the network pathways linking thiol isomerases, platelet receptor activation and fibrin generation. Several proteins, including vitronectin, thrombospondin and Factor V, have been identified as forming covalent disulfide intermediates with PDI. We are currently exploring PDI as an antithrombotic target using isoquercetin and quercetin 3-rutinoside, inhibitors of PDI identified by high throughput screening. Tail bleeding times are equivalent for mice treated with quercetin-3-rutinoside and isoquercetin compared to untreated mice. In an in vivo mouse pulmonary embolism model, PDI inhibitors rescue a high percentage of mice from death. The b-domain of PDI binds to quercetin-3-rutinoside, and infusion of the isolated b’ domain into a mouse treated with quercetin-3-rutinoside restores thrombus formation. This suggests a method for reversal of bleeding if these PDI inhibitors are found to be complicated by bleeding. The antithrombotic properties of quercetin and isoquercetin in humans have been tested. A pharmacokinetic study with quercetin and isoquercetin determined optimal oral delivery with isoquercetin. The effectiveness of these PDI inhibitors in human studies is being evaluated in a clinical trial evaluating prophylaxis of thromboembolic events in patients with cancer-associated thrombosis. PDI is a novel target for antithrombotic therapy and is unique in that its inhibition simultaneously blocks platelet thrombus formation and fibrin generation. Disclosures Zwicker: Quercegen Pharma: Research Funding.

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