thromboplastin and Leukemia--Myeloid

The induction of macrophage procoagulant activity by specific antigens is a close in vitro correlate of delayed-type hypersensitivity (DTH) reactions and provides a quantitative and sensitive measure of cell-mediated immune responses. The reaction is mediated by lymphokines from activated T lymphocytes and is potentiated by bacterial lipopolysaccharide. Lymphokines directly activated procoagulant activity on some murine and human macrophage tumor cell line cells although others required the collaboration of T lymphocytes, in the presence of lymphokine, to express activity. Macrophages in different stages of differentiation may vary, therefore, in their response to lymphokines. Pharmacological agents which regulate DTH reactions also mediated either the ability of lymphocytes to produce macrophage procoagulant inducing factor or the response of macrophages to the factors. Histamine and an anticoagulant found on stimulated T lymphocytes may be important regulatory mediators of macrophage procoagulant activity in vivo.

Topics: Animals; Cell Line; Chemical Phenomena; Chemistry, Physical; Guinea Pigs; Humans; Hypersensitivity, Delayed; Immunity, Cellular; Leukemia, Myeloid; Lymphocyte Activation; Lymphokines; Macrophage Activation; Macrophages; Mice; Monocytes; T-Lymphocytes; Thromboplastin

Topics: Blood Coagulation Disorders; Blood Coagulation Factors; Blood Platelets; Cell Membrane Permeability; Fibrinogen; Fibrinolysis; Hemorrhagic Disorders; Humans; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Thrombocythemia, Essential; Thrombocytopenia; Thromboplastin

Cancer associated thrombosis is a well-recognized phenomenon that results in considerable patient morbidity and mortality. Malignancy conveys an increased risk for thrombosis and chemotherapy further elevates this risk. The pathophysiological mechanisms underlying this process remain poorly defined.. A human acute monocytic leukemia cell line (THP-1) was treated with commonly used anthracycline chemotherapeutics at concentrations similar to those found in the plasma of cancer patients. Cells were analyzed for tissue factor (TF) mRNA, protein, and activity. Microparticle (MP) TF activity was also measured. Phosphatidylserine (PS) exposure on cells and MPs was analyzed by flow cytometry. PS levels on MPs was also evaluated in an annexin V capture assay.. Anthracycline treatment of THP-1 cells resulted in a concentration-dependent increase in cellular TF activity without a change in TF protein, which was associated with increased PS exposure on the cell surface and apoptosis. The increase in TF activity was abolished by annexin V or lactadherin indicating that PS exposure was required. Anthracycline treatment of THP-1 cells also increased the number of TF-positive MPs.. Treatment of THP-1 cells with anthracyclines induces apoptosis and increases cellular TF activity. The increased activity required an increase in exposure of PS. Additionally, anthracyclines increase the release of TF-positive MPs from THP-1 cells. We propose that the increase in cellular TF activity in circulating leukemic cells, combined with increased numbers of TF-positive MPs, may contribute to thrombosis in cancer patients receiving chemotherapy.

Topics: Anthracyclines; Antineoplastic Agents; Cell Line; Cell Membrane; Cell-Derived Microparticles; Humans; Leukemia, Myeloid; Phosphatidylserines; Thromboplastin

Different signaling routes seem to be simultaneously triggered in leukemia, with distinct and overlapping activities. To analyze if altered signals are coordinated and to evaluate their effect on this disease, we have investigated in acute myeloid leukemia samples (AML) the expression and activation status of procoagulant/proangiogenic tissue factor receptor (TF), angiogenic protein VEGF, its cell surface receptor, KDR, and two intracellular proteins involved in their regulation: extracellular regulated kinase (ERK1/2) and nuclear factor kappa-B (NFkappaB). Significantly higher mRNA and protein levels of VEGF, KDR, and TF were found in the AML samples versus controls. Enhanced ERK phosphorylation and NFkappaB activation in most AML samples were also found. In vitro MEK/ERK and NFkappaB-binding activity blockade suppressed the constitutive expression of TF, VEGF, and KDR. Anti-TF antibody treatment significantly suppressed VEGF and KDR expression as well as ERK activation, suggesting that TF expressed by AML cells may be both a regulatory target and a mediator of tumor-associated angiogenesis. Patients showing parallel activation of the studied proteins trended to exhibit higher incidence of fatal outcome. Our results show a coordinated deregulation of cellular receptors, proangiogenic factors, and intracellular pathways in leukemia cells, which may help to design mechanism-based combinations of single transduction-related therapies.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Angiogenic Proteins; Case-Control Studies; Child, Preschool; Extracellular Signal-Regulated MAP Kinases; Female; Gene Expression Regulation, Leukemic; Humans; Leukemia, Myeloid; Male; Middle Aged; NF-kappa B; Prognosis; Receptors, Cell Surface; Signal Transduction; Thromboplastin; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-2

Disturbances in the regulation of the balance between the fibrinolytic and procoagulant properties of leukemic cells may contribute to the coagulopathy of acute leukemia. The coagulant response to a number of stimuli is regulated by the expression of tissue factor, but the role of the plasminogen activator inhibitors, PAI-1 and PAI-2, in contributing to the net coagulant response is not known. In this study, we have examined the production of these proteins by cultured myeloid leukemic cells arrested at different stages of differentiation. Northern blot analysis showed time-dependent and differential production of mRNA for PAI-2 and tissue factor, and to a much lesser extent, PAI-1, in response to the differentiating agent, 12-phorbol-13-myristate acetate. The capacity to synthesize PAI-2 appeared to be related to the stage of myeloid cell differentiation. Examination of the gene products by immunoblot analysis demonstrated multiple forms of PAI-2 in all myeloid cells examined. In addition, a common characteristic of all the myeloid cells was the production of a high molecular weight species of tissue factor which may be a secreted form unique to leukemic cells. Taken together, the findings demonstrate that myeloid leukemic cells are capable of generating a multicomponent coagulant response.

Topics: Cell Differentiation; Gene Expression; Hematopoiesis; Humans; In Vitro Techniques; Leukemia, Myeloid; Plasminogen Activator Inhibitor 1; Plasminogen Activator Inhibitor 2; RNA, Messenger; RNA, Neoplasm; Tetradecanoylphorbol Acetate; Thromboplastin; Tumor Cells, Cultured

Tissue factor (TF) is transiently expressed in human monocytes exposed to the inflammatory agonist bacterial lipopolysaccharide (LPS). Since TF is the major cellular initiator of the coagulation protease cascades, it is inferred that its expression within the vasculature is strictly regulated. In this study, we investigated mechanisms which control TF mRNA expression in the human monocytic cell line THP-1. LPS induced a rapid and transient accumulation of the mature 2.2-kb TF mRNA, which was maximal at 2 h. After stimulation, the rate of transcription of the TF gene was increased (3.3 +/- 1.3)fold. In addition, we observed a significant change in TF mRNA stability: at 1 h after LPS stimulation, TF mRNA was stable during a 60-min period and had a half-life of greater than 120 min, whereas at 2 h, the half-life had declined to 25 +/- 5 min. Furthermore, a larger (3.4-kb) TF RNA species was induced in these cells; the size of this species and data from selective hybridizations with intron-specific probes are consistent with the presence of an unspliced copy of intron 1. These results demonstrate that the LPS-induced accumulation of TF mRNA levels in these monocytic cells is accomplished by both transcriptional and posttranscriptional control mechanisms.

Topics: Cycloheximide; Gene Expression Regulation; Humans; Kinetics; Leukemia, Myeloid; Lipopolysaccharides; RNA Processing, Post-Transcriptional; RNA, Messenger; Thromboplastin; Transcription, Genetic; Tumor Cells, Cultured

We investigated the induction of tissue factor by lymphokines in human monoblastic leukemia cell lines (U937) and leukemic cells from AML (acute myelogenous leukemia) patients. After incubation for 24 h, IL-2 enhanced the intracellular tissue factor 15-fold with U937 cells, and GM-CSF enhanced it 6-fold. In contrast, other lymphokines, such as IL-1-alpha, IL-1-beta, IL-3, IL-4 and G-CSF, did not affect the activity of tissue factor. The leukemic blasts, depleted of T-lymphocytes, taken from five out of 16 AML patients showed a 2.5-14-fold increase in the activity of tissue factor per cell following incubation with 200 u/ml of IL-2 for 72 h. The IL-2 induced tissue factor activity more markedly than GM-CSF. Tissue factor stimulation by IL-2 did not correlate with the expression of the IL-2 receptor, Tac, but correlated well with FAB classification of AML cells. IL-2 responders were found in M4 and M5 subtypes only, but not all M4/M5 leukemias responded to IL-2. These findings indicate that IL-2 can mediate the tissue factor induction in the monocytic type of AML and the effect is not mediated by Tac receptors. This may shed a new light on our understanding of hypercoagulability in acute monoblastic leukemia.

Topics: Cytokines; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-2; Kinetics; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Receptors, Interleukin-2; Thromboplastin; Tumor Cells, Cultured

Topics: Acute Disease; Adolescent; Adult; Aged; Blood Platelets; Clot Retraction; Female; Hematopoiesis; Humans; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid; Male; Megakaryocytes; Middle Aged; Platelet Count; Thromboplastin

Topics: Acute Disease; Animals; Aprotinin; Blood Coagulation; Chromatography, Gel; Disseminated Intravascular Coagulation; Fibrinolysis; Heparin; Humans; Leukemia, Lymphoid; Leukemia, Monocytic, Acute; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Rabbits; Thrombin; Thromboplastin; Tranexamic Acid; Trypsin Inhibitors

Topics: Cell-Free System; Hot Temperature; Humans; Hydrogen-Ion Concentration; Leukemia, Myeloid; Leukocytes; Thromboplastin

Topics: Anemia, Sideroblastic; Blood Coagulation; Blood Platelets; Humans; Leukemia, Lymphoid; Leukemia, Myeloid; Nucleotides; Platelet Adhesiveness; Primary Myelofibrosis; Thromboplastin

Topics: Adolescent; Adult; Aged; Antineoplastic Agents; Blood Coagulation Tests; Blood Platelets; Female; Humans; Leukemia, Erythroblastic, Acute; Leukemia, Myeloid; Male; Middle Aged; Myeloproliferative Disorders; Platelet Adhesiveness; Polycythemia Vera; Primary Myelofibrosis; Thromboplastin

Topics: Blood Coagulation; Blood Coagulation Tests; Heparin Antagonists; Humans; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Leukocytes; Prothrombin; Thromboplastin; Tissue Extracts

Topics: Blood Coagulation Tests; Eosinophils; Heparin Antagonists; Humans; Leukemia, Myeloid; Prothrombin; Thrombelastography; Thrombin; Thromboplastin; Tissue Extracts

Topics: ABO Blood-Group System; Adolescent; Adult; Aged; Anemia, Aplastic; Blood Coagulation; Blood Coagulation Tests; Blood Transfusion; Capillary Fragility; Centrifugation; Female; Fibrinolysin; Fibrinolysis; Gastrointestinal Hemorrhage; Hematuria; Hemostasis; Hepatitis; Humans; Leukemia, Myeloid; Male; Middle Aged; Prothrombin Time; Stomach Neoplasms; Stomach Ulcer; Thrombin; Thromboangiitis Obliterans; Thromboplastin