Tissue factor in the antiphospholipid syndrome

Antiphospholipid (aPL) antibodies are clinically important acquired risk factors for thrombosis and pregnancy loss and are thought to have a direct prothrombotic effect in vivo. Data suggest that a major mechanism by which aPL antibodies contribute to thrombophilia is the upregulation of tissue factor (TF) (CD142) on blood cells and vascular endothelium. TF is the physiological trigger of normal blood coagulation and thrombosis in many hypercoagulable conditions. This article reviews the physiology of TF, the molecular regulation of TF expression and the effects of aPL antibodies on intravascular TF regulation and expression. Inhibition of TF and the pathways by which aPL antibodies induce TF expression are potentially attractive therapeutic targets in the antiphospholipid syndrome.     

Tissue factor and inhibitor of the blood coagulation pathway in nephrotic syndrome

Some parameters of extrinsic coagulation pathway, vitronectin and thrombomodulin were studied in 21 patients with nephrotic syndrome. Concentrations of tissue factor and tissue factor pathway inhibitor were found to be significantly elevated in patients with nephrotic syndrome when compared to control group. Activities of tissue factor and tissue factor pathway inhibitor as well as vitronectin did not differ significantly from the healthy volunteers. Activities of factor VII and X were significantly higher in nephrotic patients when compared to the control group. Thrombomodulin--a marker of endothelial cell injury was significantly higher in patients with nephrotic syndrome relative to controls.     

Novel considerations in the pathogenesis of the antiphospholipid syndrome: involvement of the tissue factor pathway of blood coagulation

The antiphospholipid syndrome (APS) is characterized by clinical manifestations such as venous and arterial thrombosis, thrombocytopenia and/or recurrent pregnancy loss, as well as the persistent presence of laboratory markers of antiphospholipid (aPL) antibodies detected in laboratory assays. Though it is generally accepted that aPL antibodies, such as anticardiolipin (aCL), anti-beta2 glycoprotein I (anti-beta2GPI), and lupus anticoagulants (LA) contribute to the pathogenesis of APS, precise mechanism(s) are yet to be fully described. It is probable that aPL antibodies bind to a range of cellular targets (e.g., platelets, endothelial cells, and monocytes), leading to thrombosis and obstetric complications. There is now increasing evidence that alterations to the tissue factor (TF) pathway of blood coagulation contribute toward hypercoagulability in patients with aPL antibodies. This article reviews current evidence that suggests changes and/or interference to the major pathway of blood coagulation may represent a novel mechanism that contributes to the development of APS.     

Tissue factor in antiphospholipid antibody-induced pregnancy loss: a pro-inflammatory molecule

Fetal loss in patients with antiphospholipid antibodies (aPL) has been ascribed to thrombosis of placental vessels. However, we have shown that inflammation, specifically complement activation with generation of the anaphylotoxin C5a, is an essential mediator of fetal injury. We have analysed the role of tissue factor (TF) in a mouse model of aPL-induced pregnancy loss. TF is the major cellular activator of the coagulation cascade but also has cell signaling activity. Mice that received aPL-IgG showed strong TF staining throughout the decidua and on embryonic debris. This TF staining was not associated with either fibrin staining or thrombi in deciduas. The absence of fibrin deposition and thrombi suggests that TF-dependent activation of coagulation does not mediate aPL-induced pregnancy loss.We found that either blockade of TF with a monoclonal antibody in wild type mice or a genetic reduction of TF prevented aPL-induced inflammation and pregnancy loss indicated a pathogenic role for TF in aPL-induced pregnancy complications. In response to aPL-generated C5a, neutrophils express TF potentiating inflammation in the deciduas and leading to miscarriages. Importantly, we showed that TF in myeloid cells, but not fetal-derived cells (trophoblasts), was associated with fetal injury, suggesting that the site for pathologic TF expression is neutrophils. We found that TF expression in neutrophils contributes to respiratory burst and subsequent trophoblast injury and pregnancy loss induced by aPL. The identification of TF, acting as an important pro-inflammatory mediator in aPL-induced fetal injury, provides a new target for therapy to prevent pregnancy loss in the aPL syndrome.     

Monitoring oral anticoagulation may require determination of single coagulation factor activities in patients with antiphospholipid syndrome

We describe a 32-year-old man with secondary antiphospholipid syndrome (APS) who received oral anticoagulation with phenprocoumon after deep vein thrombosis. Conventional monitoring of oral anticoagulation by INR measurement was impaired by coagulation factor inhibition in vitro due to a strong lupus anticoagulant. The case illustrates that monitoring of oral anticoagulation may require determination of single coagulation factor activities in selected patients with APS.     

Prevalence of secondary hematologic disorders in the antiphospholipid syndrome: Impact on coagulation risk

Hematologic abnormalities may occur in antiphospholipid antibody syndrome. The most common of these abnormalities is thrombocytopenia. The mechanism by which low platelet counts occur is not known but studies continue to elucidate the potential role of autoantibodies and cell surface markers. The risk for thrombosis in patients with thrombocytopenia does not appear to be related to the presence of thrombocytopenia itself. Innovative treatment with anti-CD20 antibodies may be of benefit in reversing thrombocytopenia and treatment with splenectomy does not appear to increase the risk for subsequent thrombosis. Hemolytic anemia is associated with antiphospholipid antibodies as well, but the risk for thrombosis in patients with hemolytic anemia is not clearly elevated. Diagnostic features of disseminated intravascular coagulation may co-occur with the antiphospholipid syndrome and may be associated with a catastrophic clinical presentation.     

Bronchopulmonary cancer, antiphospholipid syndrome and coagulation disorders]

INTRODUCTION: The antiphospholipid syndrome includes recurrent thrombotic manifestations related to antiphospholipid antibodies. Adrenal insufficiency is a rare complication of the antiphospholipid syndrome. EXEGESIS: We report a case of acute adrenal insufficiency secondary to bilateral adrenal hemorrhage in a 45-year-old man. The finding of antiphospholipid antibodies and 6 months later of a polymetastatic bronchopulmonary cancer led us to diagnose a paraneoplasic antiphospholipid syndrome. CONCLUSION: We discuss the role of coagulation disorders in the pathogenesis of tumor growth and rapid metastatic spread. Assessment of the high risk for thrombosis may be of prognostic and therapeutic value in patients with evolutive bronchopulmonary cancer. Early anticoagulation treatment in association with classical treatment of cancer may contribute to prevent malignant process from extending and avoid metastatic spread.     

Synthesis of coagulation factor V by cultured aortic endothelium

Bovine aortic endothelium has been examined with respect to the synthesis of coagulation factor V. After cultured cells reached confluency, samples of supernatant culture media and solubilized cells were analyzed for factor V in a two-stage bioassay and in a double- antibody radioimmunoassay. In addition, preconfluent cells were pulsed for 4 days with 35S-methionine in methionine-free media. After the 4- day pulse, supernatant media were chromatographed on a factor V monoclonal antibody-Sepharose resin to isolate 35S-labeled factor V. The isolated material and 125I-factor V standards were analyzed by electrophoresis and autoradiography. The bioassay indicated an increase, with time, of unactivated factor V in the culture supernatant, whereas solubilized cells were negative for factor V. The radioimmunoassay indicated an increase, with time, of factor V antigen in the culture supernatants, and the solubilized cells yielded a constant level of antigen per cell. Autoradiograms of electrophoretograms of immunoadsorbed 35S-culture supernatant with 125I- factor V/Va standards revealed labeled proteins with electrophoretic mobilities compatible with 125I-factor V/Va standards. The data obtained from three different sources-bioassay, radioimmunoassay, and 35S-methionine incorporation-all indicate that factor V is synthesized by cultured bovine aortic endothelium.     

Fluctuations in levels of antiphospholipid antibodies and increased coagulation activation markers in normal and heparin-treated antiphospholipid syndrome pregnancies

Antiphospholipid antibodies (aPL) are associated with an increased risk of thrombosis and recurrent miscarriage. We assessed levels of coagulation activation markers and aPL during normal pregnancy and in women with the antiphospholipid syndrome (aPS). Fluctuations in aPL levels were observed in all patients with aPS. No particular pattern of antibody positivity, or fluctuation in aPL level, was associated with poor pregnancy outcome. A significant increase was observed in levels of factor Xlla (FXIIa; P < 0.001), factor VIIa (FVIIa, P < 0.001), thrombin antithrombin complexes (TAT; P < 0.001), prothrombin fragment F1.2 (F1.2; P < 0.001) and D-dimer (DD; P < 0.05) during normal pregnancy. Factor VIIa, TAT, F1.2 and DD increased significantly before 20 weeks gestation, while a statistically significant increase in FXIIa levels was first detected between weeks 20 and 30 of gestation. In pregnant women with aPS, increases in FXIIa were similar to those in normal pregnancy, but increased FVIIa levels were not observed until after 30 weeks gestation. Similar to normal pregnancy, increased levels of TAT and F1.2 were detected in aPS pregnancies before 20 weeks gestation, but increased DD were not observed until after week 20. Surprisingly, women with aPS receiving low molecular weight heparin prophylaxis had significantly higher (P = 0.02) levels of TAT (median 8.6; interquartile range (IQR) 6.5-20.8) between weeks 20 and 30 of gestation compared to the normal pregnant population (median 5.9; IQR 4.7-7.9), thus indicating increased thrombin generation in women with aPS in mid-pregnancy.     

Reduced factor XII levels in patients with the antiphospholipid syndrome are associated with antibodies to factor XII

Antibodies to factor XII (FXII) have previously been identified in some patients who were lupus anti-coagulant-positive. The relationship between these antibodies and FXII levels appeared to be variable. The aim of the present study was to confirm the presence of antibodies to FXII in patients with well characterized antiphospholipid syndrome (APS) and to establish their potential effect on levels of FXII. Forty-two patients with APS were studied; 21 patients were found to have either immunoglobulin (Ig)G or IgM antibodies to FXII by enzyme-linked immunosorbent assay (ELISA) using a highly purified preparation of FXII (> 99% pure). Levels of FXII were statistically significantly lower (P = 0.02) in patients with antibodies to FXII when compared with patients without antibodies to FXII (median = 91 micro/dl, s.d. = 39.1, median = 122 micro/dl, s.d. = 41.1 respectively). Four of the 21 patients with antibodies to FXII were found to have FXII levels below the laboratory normal range. Antibodies to FXII are present in significant numbers of patients with APS and may lead to acquired FXII deficiency.     

Disseminated intravascular coagulation in catastrophic antiphospholipid syndrome: clinical and haematological characteristics of 23 patients

BACKGROUND: Disseminated intravascular coagulation (DIC) is an acquired syndrome characterised by formation of microthrombi and fibrin deposition in the microvasculature. The catastrophic antiphospholipid syndrome (APS) is characterised by multiorgan thrombosis, mainly involving small vessels. A broad spectrum of disorders may develop DIC features; however, the catastrophic APS has not previously been recognised as a cause of DIC. OBJECTIVE: To analyse the clinical and laboratory characteristics of catastrophic APS patients with DIC features. METHODS: The web site based international registry of patients with catastrophic APS (CAPS registry) (http://www.med.ub.es/MIMMUN/FORUM/CAPS.HTM) was analysed and the cases with DIC features selected. RESULTS: In 173 patients with catastrophic APS, 23 (13%) were found with DIC features. The clinical and immunological characteristics were similar in catastrophic APS patients with and without DIC features; a significant difference was found only in the prevalence of thrombocytopenia (100% in patients with DIC features v 59% in those without DIC features). CONCLUSIONS: DIC features are not rare in catastrophic APS, supporting the need for systematic screening of antiphospholipid antibodies in all patients with DIC features without precipitating factors. The presence of DIC features in the context of an APS makes it imperative to rule out the catastrophic variant of this syndrome.     

Tissue factor expression determines tumour cell coagulation kinetics

Introduction: Cancers are associated with varying degrees of an increased risk of venous thromboembotic events (VTE) occurring. This increased risk is tumour driven and associated with tumour expression of tissue factor (TF) and tumour‐derived microparticles (MP). In this study, cancer cell lines from phenotypically distinct tumours were assessed for cell surface TF expression and prothrombin time (PT) taken as a measure of procoagulant potential. Methods: Breast (T47D, MCF‐7), colorectal (Colo320 and LoVo), head and neck (USCC 11b, 12, 81b and SIHN‐011A) and pancreatic tumour cell lines (ASPC‐1 and CFPAC‐1) were assessed for TF expression by flow cytometry and relative mean fluorescence determined. Procoagulant potential of the cells was then determined by PT assay. Results: Cell‐supported coagulation was shown to be cell number dependent, defined by a logarithmic relationship that was consistent across all cell lines. Single cell PT was determined for each cell line from the slope of a logarithmically transformed data plot. A near linear relationship was observed between TF expression and single cell clotting time where a higher expression of TF results in a proportionally faster PT (P < 0.001). Conclusions: This study shows that across a range of tumour sites a consistent relationship is seen between procoagulant potential and both cell number and TF cell surface expression.     

HIV infection and antiphospholipid antibody: literature review and link to the antiphospholipid syndrome

There is a high incidence of antiphospholipid antibodies, detected by assays for anticardiolipin or lupus-like anticoagulant, in HIV disease. However, a link to the antiphospholipid syndrome, with clinical thrombosis, is tenuous. We report a case of a 25-year-old man with undetermined risk factors for HIV presenting with possible antiphospholipid syndrome manifesting as necrotic skin lesions as the initial clinical presentation for HIV. We also review the literature exploring the association between HIV and antiphospholipid syndrome.     

New products for managing inhibitors to coagulation factors: a focus on recombinant factor VIIa concentrate

The treatment of alloantibody and autoantibody inhibitors directed against the factor VIII coagulant protein is one of the most challenging and expensive problems in hematology. Because the currently available plasma replacement products used in this context do not control the bleeding complications in all patients, and because of the usual emergent quality of the bleeding complications, there has been a definite need to have a uniformly reliable product for instant use, which possesses a high degree of hemostatic reliability and safety. The recent introduction of recombinant factor VIIa (rFVIIa) has been a welcome addition to the pharmacologic armamentarium for the treatment of neutralizing antibodies against coagulation factors. The mechanisms of action of rFVIIa have also been interesting and have provided insight into how the coagulation pathway accomplishes adequate hemostasis. This review will discuss this new medication and place into the context of coagulation inhibitor therapy.