thromboplastin and Hemolytic-Uremic-Syndrome

Thrombocytopenia-associated multiple organ failure (TAMOF) is a clinical phenotype that encompasses a spectrum of syndromes associated with disseminated microvascular thromboses, such as the thrombotic microangiopathies thrombotic thrombocytopenic purpura/hemolytic uremic syndrome (TTP/HUS) and disseminated intravascular coagulation (DIC). Autopsies findings in TTP, HUS, or DIC reveal specific findings that can differentiate these 3 entities. Von Willebrand factor and ADAMTS-13 play a central role in TTP. Shiga toxins and the complement pathway are vital in the development of HUS. Tissue factor is the major protease that drives the pathology of DIC. Acute kidney injury (AKI) is a common feature in patients with TAMOF.

Topics: Acute Kidney Injury; ADAM Proteins; ADAMTS13 Protein; Antibodies, Monoclonal, Humanized; Complement Inactivating Agents; Complement System Proteins; Disseminated Intravascular Coagulation; Hemolytic-Uremic Syndrome; Humans; Multiple Organ Failure; Purpura, Thrombotic Thrombocytopenic; Shiga Toxins; Thromboplastin; von Willebrand Factor

Topics: Adenocarcinoma; Anemia, Hemolytic; Animals; Blood Coagulation; Disseminated Intravascular Coagulation; Erythrocyte Aging; Erythrocytes, Abnormal; Fibrinogen; Hemolytic-Uremic Syndrome; Heparin; Humans; Hypertension, Malignant; Iodine Radioisotopes; Kidney Transplantation; Liver Transplantation; Purpura, Thrombotic Thrombocytopenic; Rabbits; Thromboplastin; Transplantation, Homologous; Venoms

Topics: Apoptosis; Biomarkers; Child; Child, Preschool; Disease Susceptibility; Endothelial Cells; Female; Heme; Heme Oxygenase-1; Hemolytic-Uremic Syndrome; Humans; Infant; Male; Oxidation-Reduction; Phenotype; Protein Transport; Reactive Oxygen Species; Shiga-Toxigenic Escherichia coli; Stress, Physiological; Thromboplastin

Endothelial expression of tissue factor (TF) may play a major role in (Stx)-related hemolytic uremic syndrome. We examined human umbilical vein endothelial cell (HUVEC) monolayers to determine the interaction between TF and TF pathway inhibitor (TFPI), hypothesizing that changes in TFPI modulate TF expression.. We studied 1) cell surface expression of globotriasylceramide (Gb3, the receptor for Stx) with Stx-1 (10 pM), TNFα (20 Ng/ml), or Stx-1 plus TNFα compared to control, 2) gene expression of TF and TFPI, 3) total cellular and cell surface antigenic TF and TFPI, 4) TFPI secretion into supernatant, and 5) factor Xa production.. Gb3 expression, negligible with control and Stx-1 alone, increased significantly with TNFα and with Stx-1 plus TNFα. TF mRNA increased 1.25 ± 0.32- fold (N = 9; p = 0.041) with Stx-1 alone vs. 2.82 ± 0.92-fold (N = 13; p < 0.0005) with TNFα alone. However, Stx-1 plus TNFα yielded a 6.51 ± 3.48-fold increase (N = 17; p < 0.0005). TFPI mRNA decreased with TNFα (p < 0.001) and Stx-1 plus TNFα (p < 0.0005). Total cellular and cell surface TF antigen increased significantly with TNFα, but no further with Stx-1 plus TNFα. Total TFPI cellular and cell surface antigen levels, and TFPI secretion decreased significantly with Stx-1 plus TNFα. Median factor Xa production for Stx-1 plus TNFα vs TNFα alone increased (p < 0.001) 3.24-fold. Our results indicate that a subinhibitory concentration of Stx-1 plus TNFα impairs TFPI gene expression, synthesis, cell-surface association, and secretion, leading to augmented functional TF.

Topics: Antigens, Tumor-Associated, Carbohydrate; Blood Coagulation; Gene Expression Regulation; Hemolytic-Uremic Syndrome; Host-Pathogen Interactions; Human Umbilical Vein Endothelial Cells; Humans; Lipoproteins; RNA, Messenger; Shiga Toxin; Shigella dysenteriae; Signal Transduction; Thromboplastin; Tumor Necrosis Factor-alpha

Aggregates formed between leukocytes and platelets in the circulation lead to release of tissue factor (TF)-bearing microparticles contributing to a prothrombotic state. As enterohemorrhagic Escherichia coli (EHEC) may cause hemolytic uremic syndrome (HUS), in which microthrombi cause tissue damage, this study investigated whether the interaction between blood cells and EHEC virulence factors Shiga toxin (Stx) and lipopolysaccharide (LPS) led to release of TF.. The interaction between Stx or LPS and blood cells induced platelet-leukocyte aggregate formation and tissue factor (TF) release, as detected by flow cytometry in whole blood. O157LPS was more potent than other LPS serotypes. Aggregates formed mainly between monocytes and platelets and less so between neutrophils and platelets. Stimulated blood cells in complex expressed activation markers, and microparticles were released. Microparticles originated mainly from platelets and monocytes and expressed TF. TF-expressing microparticles, and functional TF in plasma, increased when blood cells were simultaneously exposed to the EHEC virulence factors and high shear stress. Stx and LPS in combination had a more pronounced effect on platelet-monocyte aggregate formation, and TF expression on these aggregates, than each virulence factor alone. Whole blood and plasma from HUS patients (n = 4) were analyzed. All patients had an increase in leukocyte-platelet aggregates, mainly between monocytes and platelets, on which TF was expressed during the acute phase of disease. Patients also exhibited an increase in microparticles, mainly originating from platelets and monocytes, bearing surface-bound TF, and functional TF was detected in their plasma. Blood cell aggregates, microparticles, and TF decreased upon recovery.. By triggering TF release in the circulation, Stx and LPS can induce a prothrombotic state contributing to the pathogenesis of HUS.

Topics: Animals; Blood Platelets; Cell Separation; Enterohemorrhagic Escherichia coli; Female; Flow Cytometry; Hemolytic-Uremic Syndrome; Humans; Leukocytes; Lipopolysaccharides; Male; Mice; Shiga Toxin; Thromboplastin; Thrombosis

The pathogenesis of Shiga toxin (Stx)-mediated childhood hemolytic uremic syndrome (HUS) is not fully delineated, although current evidence implicates a prothrombotic state. We hypothesized that the tissue factor (TF) pathway plays a major role in the pathophysiology of HUS.. We measured cell surface TF activity in response to tumor necrosis factor-alpha (TNF-alpha) (20 ng mL(-1), 2-144 h), Stx-1 (10(-11) mol L(-1), 4-144 h), or their combination (TNF-alpha 22 h and Stx-1 for the last 0.5-4 h of TNF-alpha incubation) on human glomerular (microvascular) endothelial cells (HGECs) and human umbilical vein (macrovascular) endothelial cells (HUVECs).. We observed that while TNF-alpha caused an increase in cell surface TF activity on both cell types, the combination of TNF-alpha and Stx-1 differentially affected HGECs. On these cells, TF activity was increased further by 2.67 +/- 0.38-fold (n = 38, P < 0.001), consistent with our parallel observation that Stx-1 binds to HGECs but not to HUVECs. Anti-TF antibody abolished functional TF while anti-tissue factor pathway inhibitor antibody enhanced TF activity. Stx-1 alone did not induce TF activity on either cell type. Measurement of TF antigen levels and quantitative real-time polymerase chain reaction demonstrated that exposure to TNF-alpha markedly increased TF protein and TF mRNA for HGECs, but the exposure to the combination of TNF-alpha and Stx-1 did not increase further the amount of either TF protein or TF mRNA. We conclude that cytokine-activated HGECs, but not HUVECs, undergo a significant augmentation of cell surface TF activity following exposure to Stx, suggesting an important role for TF in the coagulopathy observed in HUS.

Topics: Cell Membrane; Cells, Cultured; Chromogenic Compounds; Endothelium, Vascular; Epithelial Cells; Factor Xa; Fibronectins; Hemolytic-Uremic Syndrome; Humans; Kidney Glomerulus; Lipoproteins; Microcirculation; Microscopy, Fluorescence; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Shiga Toxin 1; Thromboplastin; Time Factors; Tumor Necrosis Factor-alpha; Umbilical Veins; Up-Regulation

Severe deficiency of ADAMTS13 activity was recently found in patients with thrombotic thrombocytopenic purpura (TTP). The great advance associated with these basic and clinical studies on ADAMTS13 is the possible elucidation of the pathogenesis of TMA (thrombotic microangiopathy). However, the exact pathogenetic mechanism in TMA without severe deficiency of ADAMTS13 activity remains unknown due to heterogeneity of the disease. In this study, there were 7 patients with TTP, 7 with HUS, 3 with drug-induced HUS, 1 with VOD, and 1 with IVL out of 19 TMA patients with a moderate deficiency (6-70%) of ADAMTS13 activity. Five of the 7 TTP patients had a poor outcome. Plasma thrombomodulin and t-PA-PAI-1 complex levels in TMA patients with a moderate deficiency of ADAMTS13 activity were significantly higher than those in patients with a severe deficiency of ADAMTS13 activity. These data suggest that the etiology in these patients may be systemic vascular endothelial cell damage.

Topics: ADAM Proteins; ADAMTS13 Protein; Adolescent; Adult; Aged; Biomarkers; Female; Fibrin; Hemolytic-Uremic Syndrome; Humans; Lipoproteins; Male; Metalloendopeptidases; Middle Aged; Peptide Fragments; Protein Precursors; Prothrombin; Purpura, Thrombotic Thrombocytopenic; Thromboplastin

Topics: Biomarkers; Child; Escherichia coli Infections; Escherichia coli O157; Hemolytic-Uremic Syndrome; Humans; Thromboplastin

Microvascular thrombosis in the kidney plays an important role in the pathogenesis of hemolytic uremic syndrome (HUS). Tissue factor (TF), present on the vascular surface of endothelial cells, binds factor VIIa. The complex initiates the coagulating cascade by activating factors X and IX.. In cases of HUS associated with verotoxin-producing E. coli (VTEC) infection, VTEC gastroenteritis without HUS and normal controls, we measured plasma concentrations of TF and tissue factor pathway inhibitor (TFPI) to evaluate their clinical significance. In children with non-HUS chronic renal failure (CRF), the TF levels were also measured as another control group.. In the acute phase of HUS, plasma levels of TF and TFPI were significantly elevated, then returned to normal range in the recovery phase. The TF levels were closely correlated with the thrombin antithrombin-III complex, a marker of thrombin activity in circulating blood, and with creatinine clearance (Ccr). Furthermore, a positive correlation was noted between plasma TF levels and plasma soluble thrombomodulin (sTM) levels, which is a marker of endothelial cell injury. The influence of decreased excretion from damaged kidneys should be considered since a definite lot correlation was observed between plasma TF levels and Ccr in children with non-HUS CRF.. From these findings, we concluded that elevated TF circulating levels may also play an important role in blood-clotting activation observed in VTEC-HUS patients, and may also be a useful marker for renal damage.

Topics: Case-Control Studies; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Escherichia coli Infections; Escherichia coli O157; Female; Hemolytic-Uremic Syndrome; Humans; Lipoproteins; Male; Shiga Toxins; Thromboplastin

Diarrhoea-associated haemolytic uraemic syndrome (D+ HUS) is usually caused by verotoxin-producing Escherichia coli. Histology shows endothelial swelling with localised thrombus. Activation of coagulation and fibrinolysis also occurs. These facts, combined with the knowledge that recovery usually follows within weeks, led us to hypothesise that verotoxin causes localised endothelial cell activation but not injury. Markers of endothelial cell activation and injury were measured serially in 30 children with acute D+ HUS, healthy children, and children receiving chronic dialysis. Interpretation of markers was complicated by the renal dysfunction characteristic of D+ HUS. Nevertheless there was no evidence for endothelial cell injury, as soluble tissue factor levels were not increased and soluble thrombomodulin levels were lower than dialysed controls (P<0.001). In the acute phase, soluble vascular cell adhesion molecule levels were raised above normal (P<0.001), but were lower than dialysed controls (P<0.001), and soluble E-selectin levels were not significantly increased compared with normal controls (P=0.2). Hence, there was no evidence for endothelial cell damage or endothelial cell activation by the time children reached hospital; but this study did not exclude the possibility that endothelial cell activation occurred prior to hospital admission.

Topics: Adolescent; Biomarkers; Child; Child, Preschool; E-Selectin; Endothelium, Vascular; Hemolytic-Uremic Syndrome; Humans; Infant; Methotrexate; Solubility; Thrombomodulin; Thromboplastin; Vascular Cell Adhesion Molecule-1

Four patients had clinical manifestations of the hemolytic-uremic syndrome. No evidence of active intravascular coagulation was found during the acute phase of the illness, using a sensitive assay to measure soluble circulating fibrin in the plasma of these patients, three of whom developed the clinical syndrome while hospitalized for gastro-enteritis. These findings, coupled with the findings of others, suggest that either the episode of intravascular coagulation precedes the development of the clinical manifestations, or that platelet thrombosis is occurring in the absence of activation of plasma clotting factors. In any case, heparin anticoagulant therapy does not seem indicated.

Topics: Anuria; Blood Cell Count; Blood Coagulation Tests; Blood Platelets; Blood Pressure; Blood Urea Nitrogen; Carbon Radioisotopes; Child; Child, Preschool; Creatinine; Disseminated Intravascular Coagulation; Factor V; Factor VIII; Female; Fibrin; Fibrinogen; Hematuria; Hemoglobins; Hemolytic-Uremic Syndrome; Heparin; Humans; Infant; Male; Proteinuria; Prothrombin Time; Thromboplastin; Thrombosis

Topics: Blood Cell Count; Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Capillary Resistance; Child; Factor VIII; Fibrinogen; Hemolytic-Uremic Syndrome; Hemophilia A; Heparin; Hot Temperature; Humans; In Vitro Techniques; Infant; Kidney Failure, Chronic; Male; Nephrotic Syndrome; Plasminogen; Platelet Adhesiveness; Platelet Aggregation; Prothrombin Time; Serum Globulins; Thromboplastin

Topics: Blood Coagulation Tests; Child; Child, Preschool; Disseminated Intravascular Coagulation; Female; Hemolytic-Uremic Syndrome; Heparin; Humans; Infant; Infant, Newborn; Male; Thromboplastin