Profile of Thrombin Generation Assay and Thromboelastometry in Women with Moderate and Severe Preeclampsia. the Roadmap-Preeclampsia Study

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 37-38
Author(s):  
Patrick Van Dreden ◽  
Elmina Lefkou ◽  
Aurélie Rousseau ◽  
Grigorios T. Gerotziafas
Keyword(s):  
Endothelial Cell ◽  
Thrombin Generation ◽  
Cell Activation ◽  
Lag Time ◽  
Clot Formation ◽  
Endothelial Cell Activation ◽  
Thrombin Generation Assay ◽  
Formation Kinetics ◽  
The Mean ◽  
Α Angle

Introduction: Preeclampsia is a frequent vascular complication of pregnancy and figures among the major causes of maternal and neonatal morbidity and mortality. Early diagnosis and prompt, targeted treatment remain a unmet need. Hypercoagulability and endothelial cell activation are among the principal pathogenetic mechanisms in patients with preeclempsia. Development of diagnostic algorithms including clinically relevant biomarkers of hypercoagulability is expect to improve the management of preeclampsia. Among the numerous coagulation test, Global Coagulation Assays (GCA) such as thrombogram and thromboelastometry, could be of potential value for the evaluation of blood hypercoagulability. They provide information, on thrombin generation process, clot formation kinetics, clot firmness and even fibrinolysis potential. Aim: In this study we investigated the clinical accuracy of whole blood thromboelastometry (ROTEM®), and thrombin generation assay (calibrated automated thrombography: CAT® assay) to identify women with preeclampsia and we tried to compare their sensitivity. Methods: An observational retrospective case-control study was conducted. Plasma samples were collected from 84 women divided into three groups, the healthy pregnant (HP) group (n=35), the mild preeclampsia (MP) group (n=34) and the severe preeclampsia (SP) group (n=15). Thromboelastometry in whole blood was performed on ROTEM delta instrument (Tem Innovations GmbH, Werfen, Munich, Germany) with INTEM reagent. Thrombin generation triggered by PPP reagent low® (1 pM TF and 4µM phospholipid) was measured in platelet poor plasma. Thrombogram was also assessed in the presence or absence of thrombomodulin and the corresponding ration was calculated. Blood was collected at the diagnosis of preeclampsia (groups MP and SP) or at the equivalent months of pregnancy in the control group (HP). Statistical analysis was performed using the PASW Statistics 17.0.2 (SPSS Inc.) for Windows. Results: Thromboelastometry analysis showed that the clotting time (CT) was significantly longer in SP group as compared to MP and HP group. Both preeclampsia groups had longer clot formation time (CFT as compared to HP-group. MP-group had longer CFT as compared to SP-group. The α angle was significantly lower in SP-group as compared to the HP and MP groups. The maximum clot firmness was significantly higher in MP groups as compared to either HP or SP-group. The mean lysis (ML) was lower in both preeclampsia groups as compared to the HP group (Table 1). Thrombogram analysis showed that the lag-time of thrombin generation was significantly longer in both MP and SP groups as compared to HP group. Moreover, SP group showed significantly longer lag -time as compared to MP-group. Peak and the endogenous thrombin potential (ETP) were significantly higher in MP group as compared to either HP or SP groups. The mean rate index of the propagation phase of thrombin generation was not significantly different among the three groups whereas the thrombomodulin ratio for the ETP was significantly shorter in the SP-group (Table 2). Both tests showed a significant prolongation of the initiation phase of blood coagulation (reflected on CT and lag-time) in SP. The levels of clotting factors and fibrinogen were normal in all patients and none was on anticoagulant treatment. Thus, this prolongation reflects changes at the levels of TFPI and Thrmbomodulin reflecting an endothelial cell activation. ROTEM showed a decrease of the α-angle and MCF in SP group which is related with a lower platelet count in these patients. ROTEM showed enhanced fibrinolysis in both MP and SP groups Women with MP showed higher Peak and ETP than SP, MP showed higher ratio of ETP (TM+/TM-) than SP. Conclusion: The two GCA proved complementary information on the status of blood coagulation in pregnant women with preeclampsia. ROTEM provides information on clot formation kinetics and clot firmness as well as on fibrinolysis activation, which allow to differentiate SP from HP. However, the capacity of the assay for identification of patients with MP is limited. Thrombin generation assay showed a distinct profile between the three groups, which allowed differentiating the MP from HP as well as from SP. Disclosures No relevant conflicts of interest to declare.

scholarly journals Prospective Assessment of Biomarkers of Hypercoagulability in Oncological Patients and Healthcare Workers Following Vaccination Against Sars-Cov-2 with the mRNA Vaccine. the Roadmap-COVID-19-Vaccin Study

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3207-3207
Author(s):  
Patrick Van Dreden ◽  
Joseph Gligorov ◽  
Evangelos Terpos ◽  
Mathieu Jamelot ◽  
Michele Sabbah ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Tissue Factor ◽  
Platelet Activation ◽  
Research Funding ◽  
Thrombin Generation ◽  
Cell Activation ◽  
Control Group ◽  
Clotting Time ◽  
Endothelial Cell Activation ◽  
Active Cancer

Abstract Background: COVID-19 has been associated with hypercoagulability, endothelial cell injury and frequent thrombotic complications resulting both from direct effects of the virus on the endothelium and from the 'cytokine storm' resulting from the host's immune response. Since the COVID-19 vaccines have been shown to effectively prevent symptomatic infection including hospital admissions and severe disease, the risk of COVID-19-related thrombosis should be expected to (almost) disappear in vaccinated individuals. However, some rare cases of venous thrombosis have been reported in individuals vaccinated with mRNA vaccines. Thus, there is a sharp contrast between the clinical or experimental data reported in the literature on COVID-19 and on the rare thrombotic events observed after the vaccination with these vaccines. This phenomenon raised some scepticism of even some fear about the safety of these vaccines which could compromise the adhesion of the citizens in the vaccination program. Aims: We conducted a prospective observational study, to explore the impact of vaccination with the BNT162b2 (Pfizer/BioNTech) on blood hypercoagulability and endothelial cell activation and to investigate if this is modified by the presence of active cancer. Methods: In total 229 subjects were prospectively included in the study from April to June 2021. Subjects were stratified in three predefined groups: 127 vaccinated patients with active cancer (VOnco group), 72 vaccinated health care workers (VHcw group) and 30 non vaccinated health individuals (Control group). Blood samples were obtained 2 days after the administration of the first dose of BNT162b2 vaccine and collected in Vacutainer® tubes (0.109 mol/L trisodium citrate). Platelet poor plasma (PPP) was prepared by double centrifugation at 2000 g for 20 minutes at room temperature and plasma aliquots were stored at -80°C until assayed. Samples of PPP were assessed for thrombin generation (TG) with PPP-Reagent® (Thrombogram-Thrombinoscope assay with PPP-Reagent®TF 5pM), E-selectin, D-dimers, (D-Di), Tissue Factor (TFa), procoagulant phospholipid-dependent clotting time (Procag-PPL) and von Willebrand factor (vWF), thrombomodulin (TM), tissue factor pathway inhibitor (TFPI), and platelet factor 4 (PF4). All assays were from Diagnostica Stago (France). The upper and lower normal limits (UNL and LNL) for each biomarker were calculated by the mean±2SD for the control group. Results: All vaccinated subjects showed significantly increased levels of PF4 (71% >UNL, p<0.001), D-Dimers (74% >UNL, p<0.01), vWF (60% >UNL, p<0.01), FVIII (62% >UNL, p<0.01) and shorter Procoag-PPL clotting time (96% <LNL, p<0.001), as compared to controls. Thrombin generation showed significantly higher Peak (60% >UNL, p<0.01), ETP (38% >UNL, p<0.01) and MRI (66% >UNL, p<0.01) but no differences in lag-time in vaccinated subjects as compared to the control group. Vaccinated subjects did not show any increase at the levels of TFa, TFPI, TM and E-selectin in comparison with the control group. The studied biomarkers were not significantly different between the VOnco and VHcw groups. Conclusion: The ROADMAP-COVID-19-Vaccine study shows that administration of the first dose of the BNT162b2 vaccine induced significant platelet activation documented by shorter Procoag-PPL associated with increased levels of PF4. Plasma hypercoagulability was less frequent in vaccinated individuals whereas there was no evidence of significant endothelial cells activation after vaccination. Interestingly, the presence of active cancer was not associated with an enhancement of platelet activation, hypercoagulability, or endothelial cell activation after the vaccination. Probably, the generated antibodies against the spike protein or lead to platelet activation in a FcyRIIa dependent manner that results in PF4 release. The implication of the mild inflammatory reaction triggered by the vaccination could be another possible pathway leading to platelet activation. Nevertheless, vaccination does not provoke endothelial activation even in patients with cancer. The findings of the ROADMAP-COVID-19-Vaccine study support the concept administration of mRNA based vaccines does not directly cause a systematic hypercoagulability. Disclosures Gligorov: Roche-Genentech: Research Funding; Novartis: Research Funding; Onxeo: Research Funding; Daichi: Research Funding; MSD: Research Funding; Eisai: Research Funding; Genomic Heatlh: Research Funding; Ipsen: Research Funding; Macrogenics: Research Funding; Pfizer: Research Funding. Terpos: Novartis: Honoraria; Janssen: Consultancy, Honoraria, Research Funding; Genesis: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria, Research Funding; BMS: Honoraria; Amgen: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding; Sanofi: Consultancy, Honoraria, Research Funding; GSK: Honoraria, Research Funding. Dimopoulos: Amgen: Honoraria; BMS: Honoraria; Janssen: Honoraria; Beigene: Honoraria; Takeda: Honoraria.

scholarly journals Increased Concentration of Soluble CD40 Ligand in Preeclampsia

2007 ◽  
Vol 13 (2) ◽  
pp. 201-205 ◽  
Author(s):  
Inci Alacacioglu ◽  
Mehmet Ali Ozcan ◽  
Ozden Piskin ◽  
Faize Yuksel ◽  
Ahmet Alacacioglu ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Inflammatory Response ◽  
Cell Activation ◽  
Disease Onset ◽  
Cd40 Ligand ◽  
Endothelial Cell Activation ◽  
Soluble Cd40 Ligand ◽  
Future Studies ◽  
Study Population ◽  
The Mean

Preeclampsia has been associated with increased platelet activation detected before disease onset. Platelets are involved in hemostasis and also directly initiate an inflammatory response of the vessel wall. Inappropriate activation of platelets may be involved in pathogenesis in preeclampsia by promoting coagulation and thrombosis, and also as a mediator of inflammation. Platelets may release inflammatory mediators such as soluble CD40 ligand. The plasma level of soluble CD40 ligand was investigated during preeclamptic (n =20) and normal pregnancies (n = 20) to emphasize inflammatory response in preeclampsia. The mean soluble CD40 ligand levels were 1.08 ± 0.43 ng/mL in patients with preeclampsia and 0.76 ± 0.24 ng/mL in healthy pregnant women, which was statistically significant ( P = .01). To clarify whether inflammation may cause inappropriate endothelial cell activation or inappropriate endothelial cell activation may start this inflammatory response, future studies are needed in a larger study population.

scholarly journals Biomarkers of Hypercoagulability and Endothelial Cell Activation in Patients with Severe COVID-19 Admitted at the Intensive Care Unit. the Roadmap-COVID-19 Prospective Observational Study

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 5-6
Author(s):  
Patrick Van Dreden ◽  
Guillaume Voiriot ◽  
Alexandre Alabbadi ◽  
Ismail Elalamy ◽  
Loula Papageorgiou ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Thrombin Generation ◽  
Cell Activation ◽  
Moderate Increase ◽  
Healthy Individuals ◽  
Coagulation Activation ◽  
Endothelial Cell Activation ◽  
High Increase ◽  
Cellular Origin ◽  
Screening Algorithm

Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19) characterized by acute pneumonia which may progress to respiratory failure and life-threatening complications, including acute respiratory distress syndrome (ARDS) and multisystem organ failure with fatal outcome. COVID-19 should be regarded as a systemic disease involving multiple systems. COVID-19 is associated with excessive inflammation, platelet activation, endothelial dysfunction, blood coagulation activation and fibrin formation. The development of a screening algorithm for the evaluation of endothelial and blood coagulation activation is an urgent need for the optimisation of the management of the patients with severe COVID-19. To this aim biomarkers of endothelial cell activation and hypercoagulability have central place Aim: The prospective observational monocentric cohort study ROADMAP-COVID-19 study aimed to evaluate the biomarkers of hypercoagulability and endothelial cell activation which they could be used in the development of a screening algorithm in patients with COVID-19 Methods: The ROADMAP-COVID-19 study enrolled 90 consecutive patients hospitalized at Tenon University Hospital (AP-HP, Paris) between March 18th and April 30th, 2020 with confirmed SARS-CoV-2 infection admitted to the ICU with clinically deteriorated severe COVID-19 (ICU- group). Patients were assessed upon the first day to the ICU admission. The control group consisted 30 healthy individuals (C-group). All patients and controls were evaluated for, clotting times (PT and aPTT), fibrinogen, D-Dimers, antithrombin (AT), protein C (PC) and protein S (Ps) activity, clotting factors V (FV), VIII (FVIII) and XII (FXII), high molecular weight kininogen (HK), von Willebrand factor (FvW), thrombin generation assay (TG) using TF 20 pM-PPP-Reagent® (Thrombogram-Thrombinoscope from Stago), fibrin monomers (FM), free TFPI and TF activity, thrombomodulin activity (TM), TF expressing microparticles (the ratio TF/TFPI was also calculated) and Heparanase. The IL-6 levels were also measured. All patients hospitalized in conventional medical department or in ICU routinely received thromboprophylaxis with body weight adapted enoxaparin. The ratio of the mean values of each studied biomarker in the ICU and the C-group was calculated. Biomarkers were classified in four clusters (Decrease, Stable, Slight Increase, Moderate Increase and High Increase). Results :The ICU-group consisted of 102 patients; 87% of these patients were admitted to the ICU directly from the emergency department. Males were 76 out of 102 patients. Age ranged from 30 to 93 years in the W-group. The changes of the biomarkers in patients with COVID-19 admitted at the ICU as compared to healthy individuals with are shown in Table 1. The ICU patients with COVID-19 were characterized by marked increase of IL-6 followed by increase of heparanase and D-dimers. The PPL-ct showed the most important decrease. Patients showed a significant decrease in FXII levels which was associated with a moderate increase of kininogen levels. The levels of the natural coagulation inhibitors as well as the clotting times (PT and aPTT) did not significantly modify in patients as compared to the controls. The rate of the propagation phase of thrombin generation (MRI) and the Peak of thrombin were significantly decrease and the lag-time was prolonged reflecting the antithrombotic effect of treatment with LMWH. Heparanse, free TFPI, and TM showed moderate or high increase. The decreased Ratio of TF/TFPI indicated that the free TFPI release dominated over the release of soluble TF activity. Conclusion: Patients with severe COVID-19 requiring ICU admission showed inhaled inflammatory reaction, hyperocoagulability and endothelial cell activation whereas platelet activation appeared to be secondary. Among the studied biomarkers the PPL-ct D-dimers and heparanase followed by TFPI, FvW, FM and TM levels appear to be mandatory for hypercoagulability of cellular origin. Interestingly patients with severe COVID-19 showed consumption of FXII indicating intrinsic clotting pathway activation and high levels of HK which also indicates endothelial cell activation. The clinical relevance of these biomarkers for early detection of hypercoagulability of cellular origin in patients with COVID-19 is being evaluated in prospective study. Disclosures No relevant conflicts of interest to declare.

scholarly journals Platelet disturbances correlate with endothelial cell activation in uncomplicated Plasmodium vivax malaria

2020 ◽  
Vol 14 (7) ◽  
pp. e0007656
Author(s):  
João Conrado Khouri Dos-Santos ◽  
João Luiz Silva-Filho ◽  
Carla C. Judice ◽  
Ana Carolina Andrade Vitor Kayano ◽  
Júlio Aliberti ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Plasmodium Vivax ◽  
Vivax Malaria ◽  
Cell Activation ◽  
Endothelial Cell Activation ◽  
Plasmodium Vivax Malaria

scholarly journals Signs of sympathetic and endothelial cell activation in the skin of patients with restless legs syndrome

2021 ◽  
Author(s):  
Melanie Bergmann ◽  
Anna Heidbreder ◽  
Ambra Stefani ◽  
Cecilia Raccagni ◽  
Elisabeth Brandauer ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Restless Legs Syndrome ◽  
Cell Activation ◽  
Endothelial Cell Activation ◽  
Restless Legs

OR17 Heme oxygenase-1 modulates HLA class I antibody-dependent endothelial cell activation

2015 ◽  
Vol 76 ◽  
pp. 15
Author(s):  
Eva Zilian ◽  
Hendry Saragih ◽  
Oliver Hiller ◽  
Abid Aljabri ◽  
Constanca Figueiredo ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Heme Oxygenase ◽  
Cell Activation ◽  
Hla Class I ◽  
Class I ◽  
Heme Oxygenase 1 ◽  
Endothelial Cell Activation

scholarly journals Dengue Virus Induces the Expression and Release of Endocan from Endothelial Cells by an NS1–TLR4-Dependent Mechanism

2021 ◽  
Vol 9 (6) ◽  
pp. 1305
Author(s):  
Carlos Alonso Domínguez-Alemán ◽  
Luis Alberto Sánchez-Vargas ◽  
Karina Guadalupe Hernández-Flores ◽  
Andrea Isabel Torres-Zugaide ◽  
Arturo Reyes-Sandoval ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Mrna Expression ◽  
Cell Lines ◽  
Cell Activation ◽  
Dengue Infection ◽  
Elevated Serum ◽  
Fold Increase ◽  
Endothelial Cell Activation ◽  
Stimulation Of

A common hallmark of dengue infections is the dysfunction of the vascular endothelium induced by different biological mechanisms. In this paper, we studied the role of recombinant NS1 proteins representing the four dengue serotypes, and their role in promoting the expression and release of endocan, which is a highly specific biomarker of endothelial cell activation. We evaluated mRNA expression and the levels of endocan protein in vitro following the stimulation of HUVEC and HMEC-1 cell lines with recombinant NS1 proteins. NS1 proteins increase endocan mRNA expression 48 h post-activation in both endothelial cell lines. Endocan mRNA expression levels were higher in HUVEC and HMEC-1 cells stimulated with NS1 proteins than in non-stimulated cells (p < 0.05). A two-fold to three-fold increase in endocan protein release was observed after the stimulation of HUVECs or HMEC-1 cells with NS1 proteins compared with that in non-stimulated cells (p < 0.05). The blockade of Toll-like receptor 4 (TLR-4) signaling on HMEC-1 cells with an antagonistic antibody prevented NS1-dependent endocan production. Dengue-infected patients showed elevated serum endocan levels (≥30 ng/mL) during early dengue infection. High endocan serum levels were associated with laboratory abnormalities, such as lymphopenia and thrombocytopenia, and are associated with the presence of NS1 in the serum.

scholarly journals Experimental endotoxemia in humans: analysis of cytokine release and coagulation, fibrinolytic, and complement pathways

Blood ◽  
1990 ◽  
Vol 76 (12) ◽  
pp. 2520-2526 ◽  
Author(s):  
SJ van Deventer ◽  
HR Buller ◽  
JW ten Cate ◽  
LA Aarden ◽  
CE Hack ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Plasminogen Activator ◽  
Cell Activation ◽  
Plasminogen Activator Inhibitor ◽  
Contact System ◽  
Initial Increase ◽  
Cytokine Release ◽  
Endothelial Cell Activation ◽  
System A ◽  
Von Willebrand

Abstract Endotoxemia was evoked by bolus injection of Escherichia coli endotoxin (2 ng/kg body weight) in six healthy subjects to investigate the early kinetics of cytokine release in relation to the development of clinical and hematologic abnormalities frequently seen in gram-negative septicemia. The plasma concentration of tumor necrosis factor (TNF) increased markedly after 30 to 45 minutes, and reached a maximal level after 60 to 90 minutes. In each volunteer, the initial increase of plasma interleukin 6 (IL-6) concentrations occurred 15 minutes after the initial TNF increase, and maximal IL-6 concentrations were reached at 120 to 150 minutes. A transient increase in body temperature and pulse rate occurred simultaneously with the initial TNF and IL-6 increases, whereas a significant decrease in blood pressure occurred after 120 minutes. These changes were proportional to the changes in TNF and IL-6 concentrations. Coagulation activation, as assessed by a rise of prothrombin fragments and thrombin-antithrombin III complexes, was noted after 120 minutes, in the absence of activation of the contact system. A two- to sixfold increase in the concentrations of tissue plasminogen activator (t-PA) and von Willebrand factor antigen indicated endothelial cell activation. This increase started at 120 and 90 minutes, respectively. The release of t-PA coincided with activation of the fibrinolytic pathway, as measured by plasmin-alpha 2-antiplasmin complexes. The fibrinolytic activity of t-PA was subsequently offset by release of plasminogen activator inhibitor, observed 150 minutes after the endotoxin injection, and reaching a peak at 240 minutes. No complement activation was detected. These results show that in humans endotoxin induces an early, rapidly counteracted fibrinolytic response, and a more long-lasting activation of thrombin by a mechanism other than contact system activation. In addition, our data suggest that endotoxin-induced leukopenia and endothelial cell activation are mediated by TNF.

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