thromboplastin and Leukemia

In the last few years, it has become clear that the processes of tumor angiogenesis, metastasis and invasiveness are highly dependent on components of the blood coagulation cascade. One of the key proteins in coagulation is tissue factor (TF). In addition, TF is also known as a mediator of intracellular signaling events that can alter gene expression patterns and cell behavior. TF significantly participates in tumor-associated angiogenesis and its expression levels have been correlated with the metastatic potential of many types of hematological malignancies. Signaling pathways initiated by both, tissue factor-activated factor VII (TF-FVIIa) protease activation of protein-activated receptors (PARs), and phosphorylation of the TF-cytoplasmic domain, appear to regulate these tumoral functions. Advances in antiangiogenic therapies and preclinical studies with TF-targeted therapeutics are hopeful in the control of tumor growth and metastasis, but continued studies on the regulation of TF are still needed. In the last few years, the use of approaches of functional genomics and proteomics has allowed the discovery of new proteins involved in the origin of the neoplasia and their participation in the development of the disease. This review attempts to establish a cellular and molecular causal link between cancer coagulopathy, angiogenesis and tumor progression in hematological malignancies.

Topics: Animals; Disease Progression; Hematologic Neoplasms; Humans; Leukemia; Neoplasm Metastasis; Neovascularization, Pathologic; Proteomics; Signal Transduction; Thromboplastin; Vascular Endothelial Growth Factor A

Topics: Biomarkers; Blood Coagulation Tests; Disseminated Intravascular Coagulation; Enzyme-Linked Immunosorbent Assay; Humans; Leukemia; Lipoproteins; Neoplasms; Specimen Handling; Thromboplastin

Topics: Adult; Anemia; Blood Transfusion, Autologous; Brain Injuries; Central Nervous System Neoplasms; Cesarean Section; Female; Humans; Intraoperative Period; Leukemia; Male; Neurosurgical Procedures; Plasmacytoma; Pregnancy; Prostatic Hyperplasia; Thromboplastin

Tissue factor (TF) exists in a cryptic form [i.e. without procoagulant activity (PCA)] in peripheral blood monocytes and quiescent tissue macrophages but is expressed constitutively in most human tumor cells. Induction and cell surface expression of TF in these cells in vivo is associated with activation of intravascular and extravascular coagulation in patients with a variety of inflammatory or malignant diseases. The regulation of TF synthesis in cells is complex and new information from transfection studies suggests that changes in cellular glycosylation pathways impair cell surface expression of functional TF. Such dysregulation may also characterize the lineage-unfaithful expression of TF in leukemic cells and perhaps explain some of the thrombohemorrhagic complications in patients with acute progranulocytic leukemia. The importance of carbohydrate modification of TF is reviewed.

Topics: Acute Disease; Animals; Blood Coagulation Disorders; Carbohydrate Sequence; Cell Differentiation; CHO Cells; Cricetinae; Cricetulus; Cysteine Endopeptidases; Glycosylation; HL-60 Cells; Humans; Leukemia; Leukocytes; Molecular Sequence Data; Neoplasm Proteins; Neoplasms; Neoplastic Stem Cells; Thromboplastin

Topics: Acute Disease; Animals; Disseminated Intravascular Coagulation; Hematologic Diseases; Heparin; Humans; Leukemia; Rabbits; Thromboplastin

Tissue factor is an ubiquitous phospholipid-protein complex, which triggers blood coagulation through the so-called extrinsic pathway. Reactions initiated by tissue factor bypass many of the early stages of coagulation (contact phase) and involve factors VII, X, V, II and fibrinogen but also factor IX (and VIII) as it was recently demonstrated. So, it appears that tissue factor has a key-role in the haemostasic process as it has been suggested by the mildness or the absence of haemorrhagic syndrome in contact factors deficiencies. Tissue factor activity has been found in many types of cells, especially in white bloods cells. Experimental studies have demonstrated the presence of tissue factor activity in polymorphonuclears, lymphocytes, monocytes (or macrophages). This activity is enhanced by gram-negative endotoxin stimulation, inflammation, cell mediated immunologic phenomena or malignancy. These data are in good agreement with a wild range of features observed in human pathology: fibrin deposits in inflammatory lesions, disseminated intravascular coagulation (DIC) during the course of gram-negative septicemias or acute promyelocytic leukemias, local thrombi at the early phase of graft rejection. The protective effect of a phospholipase C against DIC induced in rats by tissue factor infusion suggests in the future, a specific therapy would be possible in man that, in the frequent clinical conditions involving clotting activation through tissue factor pathway.

Topics: Animals; Blood Coagulation; Blood Coagulation Factors; Disseminated Intravascular Coagulation; Fibrin; Graft Rejection; Humans; Inflammation; Leukemia; Leukocytes; Rabbits; Sepsis; Thromboplastin

Topics: Adolescent; Adult; Blood Coagulation; Blood Coagulation Disorders; Child; Disseminated Intravascular Coagulation; Embolism, Fat; Endotoxins; Fatty Acids; Female; Fibrinolysis; Heart Defects, Congenital; Hemangioma; Hemolysis; Humans; Leukemia; Male; Middle Aged; Mononuclear Phagocyte System; Neoplasms; Pregnancy; Pregnancy Complications; Purpura, Thrombocytopenic; Skin Neoplasms; Thromboplastin

Topics: Antithrombins; Blood Coagulation; Fibrinolysis; Hemorrhagic Disorders; Hemostasis; Heparin Antagonists; Humans; Leukemia; Leukocytes; Thromboplastin

Topics: Blood Transfusion; Enzyme Inhibitors; Female; Fibrin; Fibrinogen; Fibrinolysin; Fibrinolysis; Hemostatics; Humans; Leukemia; Liver Cirrhosis; Male; Pathology; Physiology; Polycythemia Vera; Pregnancy; Pregnancy Complications; Pregnancy Complications, Hematologic; Prostatectomy; Prostatic Neoplasms; Prothrombin; Thrombin; Thromboplastin

Thrombin, the final enzyme of blood coagulation, is a multifunctional serine protease also involved in the progression of cancer. Tumor cells may activate blood coagulation proteases through the expression of procoagulant activities. However, specific information about the thrombin generation potential of malignant tissues is lacking. In this study we applied a single global coagulation test, the calibrated automated thrombogram assay, to characterize the specific procoagulant phenotypes of different tumor cells.. Malignant hematologic cells (i.e. NB4, HEL, and K562) or solid tumor cells (i.e. MCF-7 breast cancer and H69 small cell lung cells) were selected for the study. The calibrated automated thrombo-gram assay was performed in normal plasma and in plasma samples selectively deficient in factor VII, XII, IX or X, in the absence or presence of a specific anti-tissue factor antibody. Furthermore, cell tissue factor levels were characterized by measuring antigen, activity and mRNA expression.. In normal plasma, NB4 induced the highest thrombin generation, followed by MCF-7, H69, HEL, and K562 cells. The anti-tissue factor antibody, as well as deficiencies of factors VII, IX and XII affected the thrombin generation potential of malignant cells to different degrees, allowing differentiation of the two different pathways of blood clotting activation - by tissue factor or contact activation. The thrombin generation capacity of NB4 and MCF-7 cells was tissue factor-dependent, as it was highly sensitive to inhibition by anti-tissue factor antibody and factor VII deficiency, while the thrombin generation capacity of H69, HEL and K562 was contact activation-dependent, as no thrombin was generated by these cells in factor XII-deficient plasma.. This study demonstrates that the calibrated automated thrombogram assay is capable of quantifying, characterizing, and comparing the thrombin generation capacity of different tumor cells. This provides a useful tool for understanding the key factors determining the global pro-coagulant profile of tumors, which is important for addressing specific targeted therapy for the prevention of thrombosis and for cancer.

Topics: Blood Coagulation Tests; Cell Line, Tumor; Factor X Deficiency; Factor XII Deficiency; Hemophilia B; Humans; Leukemia; Neoplasms; Thrombin; Thromboplastin

This study examined the expressions of human serum tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in patients with acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and their clinical significance. The serum TF and TFPI levels were detected by ELISA in 28 allo-HSCT recipients before and after the transplantation and the changes of TF and TFPI levels were dynamically monitored at different phases of the disease. No significant differences in the serum TF and TFPI levels were found in allo-HSCT recipients in the absence of aGVHD or with grade I aGVHD before and after the transplantation. The levels of serum TF and TFPI were substantially increased in the patients with gradeII aGVHD at the peak of aGVHD (P<0.05) and they were even higher in the patients with grade III-IV aGVHD (P<0.01). When the conditions became stable after treatment with immunosuppressive agents, the serum TFPI level was decreased to the baseline level (P>0.05) and the TF level was lowered but still higher than the baseline level (P<0.05). It was concluded that the levels of serum TF and TFPI were increased significantly in the patients with grade II-IV aGVHD after allo-HSCT and decreased markedly after the treatment. Monitoring the levels of serum TF and TFPI in the patients with allo-HSCT is important to predict the occurrence, outcome and prognosis of aGVHD.

Topics: Acute Disease; Adolescent; Adult; Female; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Leukemia; Lipoproteins; Male; Middle Aged; Prognosis; Thromboplastin; Transplantation, Homologous; Young Adult

This study was purposed to explore the significance of tissue factor (TF), tissue factor pathway inhibitor (IFPI) and interleukin-1beta (IL-1beta) in the evaluation of development, curative effect and prognosis of AL patients. ELISA was used to detect the levels of TF, TFPI and IL-1beta in plasma of 20 healthy individuals and 24 newly diagnosed AL patients. All the three indications of patients were measured in different stages including pre-chemotherapy phase, at 72 hours after chemotherapy, complete remission phase. The results showed that as compared with normal control, levels of TF, TFPI and IL-1beta in plasma of AL patients during pre-chemotherapy phase were higher (p < 0.01); as compared with pre-chemotherapy phase, levels of TF, IL-1beta were elevated at 72 hours after -chemotherapy (p < 0.05). However, the levels of TFPI was much lower than that of 72 hours after chemotherapy (p < 0.01). 16 out of 24 patients got complete remission, there was no difference of TF, TFPI and IL-1beta between complete remission group and normal control group. It is concluded that the levels of TF, TFPI and IL-1beta in plasma can be used as the indicators for understanding clinical features, evaluating disease status and predicting prognosis in acute leukemia patients.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Child; Female; Humans; Interleukin-1beta; Leukemia; Lipoproteins; Male; Middle Aged; Thromboplastin; Young Adult

This paper presents an analysis of 24 cases in which recombinant factor VIIa (rFVIIa) was used in the management of hemorrhage in patients with thrombocytopenia associated with hematologic malignancies. This is the largest case aggregation to date and focuses on preliminary experience in the off-label use of this hemostatic agent. Data were extracted from the international, Internet-based registry, www.haemostasis.com, accessed in September 2003. The search results were manually cross-checked against monthly summary reports. The physicians providing the cases were contacted individually to approve the use of their cases, supply any information missing from the database, and validate the data already held. Patients with acute myeloid leukemia, acute lymphoblastic leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, Burkitt's lymphoma, B-cell or T-cell lymphoma, or aplastic anemia received rFVIIa at total doses of between 18 and 1040 mug/kg body weight. Bleeding stopped in 11 of 24 (46%) patients, markedly decreased in 8 of 24 (33%) patients, and decreased in 4 of 24 (17%) patients. In most patients, the response was achieved within 2.5 hours of administration of rFVIIa. The use of rFVIIa was generally well tolerated -- 1 case of ischemic stroke was considered to be possibly related to rFVIIa administration, but this has yet to be confirmed. A review of these 24 cases submitted to the www.haemostasis.com database suggests that rFVIIa is beneficial in the management of hemorrhage in patients with thrombocytopenia and hematologic malignancies. This warrants further investigation in rigorously controlled clinical trials.

Topics: Adolescent; Adult; Anticoagulants; Child; Child, Preschool; Factor VIIa; Female; Hemorrhage; Humans; Internet; Leukemia; Lymphoma; Male; Medical Audit; Middle Aged; Prothrombin; Thrombocytopenia; Thromboplastin

The aim of study was to evaluate TF activity and TFPI concentration in patients with haematological malignancies and urinary tract tumors. TFPI concentration and activity and TF concentration were measured in 20 patients suffering from acute myeloblastic leukaemia (AML), 21 patients with chronic myelogenous leukaemia (CML), 17 patients with chronic lymphatic leukaemia (CLL), 16 patients with multiple myeloma (MM) and 65 healthy adults. TFPI and TF concentrations were measured also in patients with renal cell carcinoma (n = 12) and bladder cancer (n = 17) and patients with benign prostatic hyperplasia (BPH) (n = 15). Patients with AML, CML, CLL, and cancer revealed elevated TFPI concentrations. Patients with AML, CML, CLL, MM showed decreased TFPI activity. However TFPI concentration correlated inversely with TFPI activity only in the AML group. No significant changes were observed in TF concentrations in all investigated groups.

Topics: Biomarkers, Tumor; Carcinoma, Renal Cell; Case-Control Studies; Enzyme-Linked Immunosorbent Assay; Female; Humans; Kidney Neoplasms; Leukemia; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Leukemia, Myeloid, Acute; Lipoproteins; Male; Multiple Myeloma; Prostatic Hyperplasia; Thromboplastin; Urinary Bladder Neoplasms; Urologic Neoplasms

Tissue factor (TF) antigen and activity were measured in leukemic cell homogenates. In leukemic cell homogenate, especially that of acute promyelocytic leukemia (APL), both TF antigen and activity were significantly higher than these levels in the mononuclear cells obtained from healthy volunteers. Both TF antigen and activity were significantly higher in myelocytic leukemia than in lymphocytic leukemia cells. In leukemic cell homogenates, there was a close correlation between TF antigen and TF activity. The TF activity/TF antigen ratio was significantly higher in myelocytic leukemia than in lymphocytic leukemia cells. As the TF activity was not increased in lymphocytic leukemia cell homogenates to which were added phospholipids, the decrease in TF activity in lymphocytic leukemia might not be due to phospholipid in the leukemic cell membrane. Values for TF activity, TF antigen, and the TF activity/TF antigen ratio in leukemic cell homogenate from patients with disseminated intravascular coagulation (DIC) were significantly higher than those in patients without DIC. Therefore, the measurement of TF antigen and activity in leukemic cells could be useful for the prediction of DIC.

Topics: Antigens, Neoplasm; Disseminated Intravascular Coagulation; Humans; Leukemia; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Monocytic, Acute; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Leukemia, Myelomonocytic, Acute; Leukemia, Promyelocytic, Acute; Leukemia, T-Cell; Leukocytes, Mononuclear; Neoplasm Proteins; Thromboplastin

Our previous study described a novel biologic function of compound 48/80 (48/80) in the downregulation of monocytic tissue factor (TF)-initiated hypercoagulation in response to bacterial endotoxin (lipopolysaccharide; LPS). The inhibition was not due to the blockade of LPS cell signaling, as evidenced by the unaffected LPS-induced TF synthesis. We herein determined the mechanism by which 48/80 inhibits the extrinsic coagulation in agonist-challenged THP-1 monocytes. LPS as well as A23187 substantially induced TF activity. TF synthesis was enhanced by LPS but not by A23187. However, the elevated FVII binding to monocytes accompanying the upregulation of factor VII (FVII) activation was uniformly observed in both cases. A 5-min preincubation of the cells with a sheep anti-humanTF antibody (anti-hTF Ab) showed the downregulation of FVII activation, indicating a regulatory role of FVII binding in the modulation of the extrinsic coagulation. The 48/80 blocked FVII binding to monocytes, leading to the preferential inhibition of FVII activation. As the result of the diminished FVIIa formation, monocytic TF-initiated extrinsic coagulation was downregulated in agonist-challenged THP-1 monocytes.

Topics: Blood Coagulation; Cells, Cultured; Endotoxins; Factor VII; Humans; Leukemia; Lipopolysaccharides; Monocytes; p-Methoxy-N-methylphenethylamine; Thromboplastin; Up-Regulation

In order to investigate the leukemogenic potential of PLZF-RARA fusion protein in vivo, hCG-PLZF-RARA transgenic mice were generated, in which PIZF-RARA fusion gene was driven by hCG promoter to express in myeloid cells of mice.. Molecular cloning technology was used to construct hCG-PLZF-RARA gene. The genotype and phenotype of the hCG-PLZF-RARA transgenic mice were analyzed by PCR, RT-PCR, immunofluorescence, morphology of bone marrow (BM) cells, pathology and retinoic acid differentiation assays.. Six hCG-PLZF-RARA transgenic mice developed leukemia resembling human chronic myeloid leukemia. TF(tissue factor) was not expressed in BM cells of normal mice nor in mice without the expressed transgene, but it was expressed in mice expessing the transgene.. PLZF-RARA fusion protein plays a crucial role in leukemogenesis. TF is up-regulated by PLZF-RARA fusion gene.

Topics: Animals; Cell Transformation, Neoplastic; Chorionic Gonadotropin; Disease Models, Animal; Humans; Leukemia; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Mice; Mice, Transgenic; Neoplasm Proteins; Oncogene Proteins, Fusion; Recombinant Fusion Proteins; Thromboplastin; Up-Regulation

We have recently found that retinoic acids (RAs) evoke anticoagulant effect by upregulating thrombomodulin (TM) and downregulating tissue factor (TF) expression in acute promyelocytic leukemia (APL) and monoblastic leukemia cells. Two classes of nuclear RA receptors, termed retinoic acid receptors (RARs) and retinoid X receptors, have been identified. Each receptor class consists of three subtypes. We have used several synthetic retinoids to find which receptor subtypes are involved in the regulation of TM and TF expression in APL cells NB4, monoblastic leukemia cells U937 and human umbilical vein endothelial cells (HUVECs). Am80, which does not have a binding affinity to RARgamma, Ch55, which does not bind to cytoplasmic retinoic acid binding protein (CRABP), and a specific RARalpha agonist Ro40-6055, have shown to upregulate TM and downregulate TF in NB4 and U937 cells similar to all-trans RA (ATRA). A specific RARalpha antagonist Ro41-5253 efficiently suppressed the upregulation of TM by ATRA and Am80 in NB4 cells, U937 cells and HUVECs. In contrast, only when both RARalpha and RARbeta antagonists were preincubated, downregulation of TF by the retinoids was suppressed in NB4 cells. These results indicate the mechanically distinct transactivation and transrepression functions of RARs, the major role of RARalpha in TM upregulation by retinoids in leukemic cells and HUVECs and the cooperative role of RARalpha and RARbeta in TF downregulation by retinoids. This implies that synthetic retinoids will provide a very useful means to control distinct targets, TM and TF genes, at the level of transcription. Synthetic retinoids may develop as new type of antithrombotic agents which may change the character of cells as well as act as malignant cell differentiation inducers.

Topics: Anticoagulants; Antineoplastic Agents; Benzoates; Cell Differentiation; Chromans; Dibenzazepines; Humans; Leukemia; Membrane Proteins; Retinoids; Thrombomodulin; Thromboplastin; Tretinoin; Tumor Cells, Cultured; U937 Cells

Tissue factor (TF) and urokinase receptor (uPAR) are key cellular receptors triggering, respectively, coagulation and fibrinolysis. Bleeding complications among leukemic patients have been related to an abnormal expression of TF by blast cells and/or to an abnormal fibrinolytic response. In this study the expression of TF and uPAR has been assessed in 18 acute non-lymphoblastic and 8 lymphoblastic leukemic blast cells using several methodological approaches. TF mRNA was evaluated by in situ hybridization and TF and uPAR antigen were evaluated immunologically in cell lysates and on the cell surface by flow cytometry. In addition, TF-procoagulant activity was measured in coagulation-based assays. The reliability of these methods was corroborated in six leukemic cell lines of different lineages and states of maturation. Disseminated intravascular coagulation was detected in two M3 leukemia patients whose blast cells expressed high amounts of TF. Hyperfibrinolysis was detected in one M1 and two M2 patients, whose blast cells displayed a high content of uPAR antigen, but no TF. Furthermore, M5 leukemia blast cells expressed both TF and uPAR, although no hemostatic defects or bleeding complications were detected in these patients. Taken together, although a limited number of patients was included in this study, these data suggest that in leukemia patients exhibiting bleeding, either TF or uPAR are expressed by their blast cells. However, the presence of these receptors does not necessarily imply the existence of a hemostatic disorder.

Topics: Acute Disease; Blood Coagulation; Hemorrhage; Humans; Leukemia; Receptors, Cell Surface; Receptors, Urokinase Plasminogen Activator; Thromboplastin; Tumor Cells, Cultured

We have recently found that all-trans retinoic acid (ATRA) upregulates thrombomodulin (TM) and downregulates tissue factor (TF) expression in acute myelogenous leukemia (AML) M3 cells (NB4) and acute monoblastic leukemia cells (U937) (Koyama et al, Blood 84:3001, 1994). We have further investigated the effects of ATRA on leukemic cells freshly isolated from patients at diagnosis. Increase of TM antigen was documented in all AML cells: M0 (n = 1), M2 (n = 5), M3 (n = 3), M4 (n = 3), M5 (n = 3), and M6 (n = 1). Decrease of TF antigen was observed in 4 M2, 1 M4, and all M3 and M5 patients. However, no TM and TF antigens were detected in all chronic lymphocytic leukemia cells (n = 3) with or without ATRA treatment. Changes of TM and TF antigen levels were associated with those of TM and TF cofactor levels on the cell surface. A stereoisomer of RA, 9-cis RA, is a high-affinity ligand for the RA receptors (RARs) and the retinoid X receptors, although ATRA and another isomer, 13-cis RA, solely bind to RARs. We have also studied the effects of 9-cis RA and 13-cis RA on the expressions of TM and TF in NB4 and U937 cells. A relatively wide range of 9-cis RA concentrations (0.01 to 1 mumol/L) compared with ATRA was optimal for prolongation of normal plasma-based recalcification time (reduction of cell surface TF activity), decrease of TF antigen, and increase of TM antigen on the surface and in the lysates of NB4 and U937 cells. Western blot analysis under nonreducing conditions showed that both ATRA and 9-cis RA markedly induced the prominent band at 75 kD of TM and reduced the band at 45 kD of TF. Northern blot analysis has shown similar changes of mRNA levels, which indicates that RAs regulate TM and TF expression in leukemic cells at transcriptional levels. Anticoagulant effects of ATRA, ie, upregulation of TM expression and downregulation of TF expression, are applied not only to established cell lines of specific subtypes (M3 and M5) but also to more universal AML (most cases of M3 and M5 and a part of the other types of AML) cells freshly isolated from patients. 9-cis RA may be more effective than ATRA as an inducer of differentiation of AML M3 cells and as an anticoagulant agent for patients with certain types of AML as well.

Topics: Anticoagulants; Base Sequence; Cell Separation; Cysteine Endopeptidases; Flow Cytometry; Gene Expression Regulation, Leukemic; Humans; Isotretinoin; Leukemia; Leukemia, Monocytic, Acute; Leukemia, Promyelocytic, Acute; Lymphoma, Large B-Cell, Diffuse; Molecular Sequence Data; Neoplasm Proteins; Neoplastic Stem Cells; Receptors, Retinoic Acid; Thrombomodulin; Thromboplastin; Tretinoin; Tumor Cells, Cultured

In 43 patients with leukemia, we determined the increase of tissue factor (TF) activity production by leukemic cells that was induced by incubation with endotoxin at the time of admission. Definite disseminated intravascular coagulation (DIC) developed in 8 patients on admission (Group III) and in 8 patients just after the initiation of treatment (Group II), but not in the remaining 27 patients (Group I). TF activity before incubation (TF1) was 0.70 U/10(8) cells or more in 6 patients of Group III, while it was less in all the patients of Group II and 25 of the 27 patients of Group I. On the other hand, TF activity after incubation with endotoxin (TF2) increased to more than 1.11 U/10(8) cells in all the patients of Groups II and III, while it remained less in all the patients of Group I. These results suggest that the leukemic cells in Group II might not have expressed sufficient TF activity to cause DIC until chemotherapy was begun, and that 1.11 U/10(8) cells or more of TF2 might strongly indicate the development of DIC during treatment for leukemia.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Cell Count; Disseminated Intravascular Coagulation; Endotoxins; Female; Humans; In Vitro Techniques; Leukemia; Male; Middle Aged; Risk Factors; Thromboplastin

Tissue factor activity of intact cell and cell lysate, and the presence of tissue factor antigen on cell surface, were examined in leukemic cells from patients with acute myelogenous leukemia (AML, M1-M5) or acute lymphoblastic leukemia (ALL-L1), and in mononuclear cells from normal donors. Leukemic cells from AML or ALL had significantly more tissue factor activity not only on intact cells but also in cell lysate than mononuclear cells from normal donors (p < 0.001). Tissue factor activities of the intact leukemic cells and lysate from AML patients with DIC were significantly higher than those without DIC (p < 0.001). The relationship between the percent of positive cells for tissue factor and the presence of DIC at the time of diagnosis of acute leukemia was observed. The patients with DIC showed the higher percentage of tissue factor-positive cells than those without (p < 0.01). The development of DIC following chemotherapy was recognized in 2 out of 7 AML-MI patients and 2 out of 4 ALL-L1 patients who had relatively high tissue factor activities of cell lysate. The release of tissue factor from cytoplasm induced by chemotherapy would be another mechanism for the development of DIC. The report suggests the possibility of the prediction for DIC by the flowcytometric assay of tissue factor antigen.

Topics: Antigens, Surface; Disseminated Intravascular Coagulation; Flow Cytometry; Humans; Leukemia; Leukemia, Myeloid, Acute; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Thromboplastin

We studied the effect of fibrinogen degradation products D, E, and D-dimer on a human promonocytic leukemia cell line, NOMO-1. After exposure to a 10(-5)-mol/L fragment D or D-dimer, the cells displayed macrophage-like characteristics, such as adherence to plastic surfaces, and showed approximately a twofold increase in response to the nitroblue tetrazolium reduction test. The secretion of interleukin-1 alpha (IL-1 alpha) into the medium was markedly stimulated by a 10(-5)-mol/L fragment D, E, and D-dimer, whereas a significant increase in IL-1 beta secretion was observed only in D-dimer-stimulated cells. In addition, D-dimer induced a rapid increase in urokinase-type plasminogen activator on day 1 (0.52 +/- 0.02 ng/mL v 0.07 +/- 0.01 ng/mL in the control culture) and a slow increase in plasminogen activator inhibitor-2 on day 5 (3.9 +/- 1.6 ng/mL v 1.2 +/- 0.2 ng/mL in the control culture). An increase in tissue factor (TF) was also demonstrated on the cell surface of NOMO-1 cells exposed to fragment D or D-dimer by indirect immunofluorescence using an anti-TF monoclonal antibody. Scatchard plot analysis showed that fragment D and D-dimer bound to the NOMO-1 cells with a kd of 3.3 nmol/L and 2.7 nmol/L, respectively. These results suggest that fragment D-dimer specifically stimulates cells of monocyte-macrophage lineage to secrete key substances that regulate blood coagulation, fibrinolysis, and inflammation.

Topics: Antibodies, Monoclonal; Cell Line; Enzyme Precursors; Fibrin Fibrinogen Degradation Products; Fluorescent Antibody Technique; Humans; Interleukin-1; Kinetics; Leukemia; Phagocytosis; Plasminogen Activators; Plasminogen Inactivators; Superoxides; Thromboplastin; Urokinase-Type Plasminogen Activator

It has been established that the only source of tissue thromboplastin activity in homogenate of cells (HeLa, L-41, RAMT, Hep-2 and FL) grown in synthetic nutrient media are permolecular formations represented by fragments of cellular membranes Thromboplastic activity of nutrient media is also associated with these formations, as fragments of cellular membranes enter the media during cell cultivation. The removal of all permolecular formations from homogenate of cells, from the medium in which they are cultivated, allows determination of the topography of tissue coagulation factors.

Topics: Amnion; Animals; Callitrichinae; Carcinoma, Squamous Cell; Cattle; Cell Line; Culture Media; Female; HeLa Cells; Humans; In Vitro Techniques; Kidney; Laryngeal Neoplasms; Leukemia; Male; Thromboplastin

Apoprotein part of tissue factor of human placenta was purified 871 fold from the starting material with 4.2% yield by concanavalin A-Sepharose affinity chromatography and SDS-PAGE. The molecular weight of purified apoprotein was 45,000 in non-reduced condition and 49,000 in reduced condition. Tissue factor of human leukemia cells (FAB classification:M2 and M3) and cultured leukemia cell lines (HL-60 and Molt-4) was analyzed using specific rabbit anti-tissue factor IgG raised against purified material. Endotoxin stimulated HL-60 and Molt-4 also expressed procoagulant activity which was inhibited by tissue factor immune IgG. By immunostaining of the purified material, the lysate of leukemia cells (M2 and M3) and cultured leukemia cells (HL-60 and MOLT-4) revealed a major band of the same apparent molecular weight. Immuno-electron microscopic study on tissue factor of HL-60 cells produced the following findings: stimulation by endotoxin resulted in the formation of pseudopods of the cell membrane, and immunogold particles accumulated mainly on these pseudopods and cisternal spaces of rough endoplasmic reticulum, indicating exposure of the tissue factor to the surface of perturbed cell membrane with concurrent increase in tissue factor synthesis.

Topics: Antibody Formation; Apoproteins; Blood Coagulation; Female; Humans; Immunoblotting; Leukemia; Neutralization Tests; Peptide Hydrolases; Placenta; Pregnancy; Thromboplastin; Tumor Cells, Cultured

Tissue factor activity (TFA) of 10(8) leukemia cells was measured in 82 patients with acute nonlymphoid leukemia by the clotting method. The TFA bore a significant correlation to the development of disseminated intravascular coagulation (DIC) in these cases. Mean TFA value with standard deviation (SD) was 8.3 +/- 6.3 U in 48 cases with DIC, which was significantly higher than 0.3 +/- 4.2 U in 34 cases without DIC. Whereas Mean TFA in non-M3 was 0.9 +/- 6.3 U which was significantly lower than 37.2 +/- 2.3 U in M3, some non-M3 showed TFA as high as M3 and were complicated by DIC. In heparin treatment, dosage of heparin could not be controlled by either APTT or AcCT but was controlled by the extent of TFA of leukemia cells. Retrospective analysis of clinical features revealed that 97000X + 9000 units/day (X = logarithm value of TFA) of heparin is an adequate dosage for the successful treatment of DIC when TFA of leukemia cells is 0.8 U or more.

Topics: Adolescent; Adult; Aged; Antigens; Disseminated Intravascular Coagulation; Female; Heparin; Humans; Leukemia; Male; Middle Aged; Retrospective Studies; Thromboplastin

Topics: Cell Line; Cell Membrane; Factor X; Hemostasis; Humans; Leukemia; Leukocytes; Thromboplastin; Thrombosis

The apoprotein (AP) of tissue factor (TF) has been purified 72,000-fold to homogeneity from human placenta using acetone delipidation, sodium deoxycholate (DOC) extraction, Sephacryl S-300 column chromatography, preparative polyacrylamide-gel electrophoresis (PAGE) in DOC and tryptic digestion. The purified AP had an apparent molecular weight of 54,000 by sodium dodecyl sulfate/PAGE. A radioimmunoassay (RIA) for quantitation of the TF-AP using an antibody against this purified AP of TF was devised which was sensitive enough to measure as small a quantity as 100 pg/ml of TF-AP accurately with high reproducibility. In addition to TF clotting activity (TFA), the immunoreactive TF-AP (IR-TFR) in the homogenates of leukemic leukocytes from patients with acute non-lymphoid leukemia (ANLL) was determined using this RIA. In 30 patients with ANLL, the mean IR-TFR with standard deviation (SD) of 21 cases with DIC was 157.9 +/- 188.1 ng/10(8) cells, which was significantly higher than that (37.1 +/- 29.9 ng/10(8) cells) of 9 cases with no DIC during remission induction chemotherapy (p less than 0.01).

Topics: Apoproteins; Disseminated Intravascular Coagulation; Female; Humans; Leukemia; Molecular Weight; Placenta; Pregnancy; Radioimmunoassay; Thromboplastin

The procoagulant cellular activity (PCA) of intact and lysed leukaemic cells was evaluated at diagnosis in 23 patients with acute non-lymphoid leukaemia (ANLL). The leukaemic cells of all 13 patients having DIC feature (excess of fibrin monomers, serum FDP and plasma fibrino-peptide A) showed a significant (P less than 0.0001) increase of PCA, while a pattern similar to that of normal granulocytes and lymphomonocytes was observed in the remaining 10 patients without evidence of DIC. When the patients were subdivided according to the FAB cytological classification, features of DIC and increased PCA were demonstrated in 3/3 M3 patients, 5/6 M5 patients and only in 5/14 remaining patients. These findings indicate that in ANLL patients: (1) the increased PCA of leukaemic cells is closely related to the occurrence of DIC; (2) the increased PCA seems related to the differentiation line and maturation level of the leukaemic cells.

Topics: Acute Disease; Adolescent; Adult; Aged; Blood Coagulation Tests; Bone Marrow; Disseminated Intravascular Coagulation; Female; Granulocytes; Hematopoietic Stem Cells; Humans; Leukemia; Lymphocytes; Male; Middle Aged; Monocytes; Thromboplastin

Procoagulant activity of gastric cancer tissues and leukocytes obtained from various types of leukemia have been studied with special reference to TTP. The following results were obtained. Homogenates of APL leukocytes and gastric cancer tissues contained strong procoagulant activities, most of which have been identified as TTP since the activities were neutralized by a specific antibody against purified human placenta TTP, inactivated by the removal of phospholipid with heptane-butanol mixture, and inactivated by the addition of phospholipase C. The delipidated homogenates regained procoagulant activities by relipidation procedures. These results also confirmed that TTP from APL leukocytes and gastric cancer tissues have the same lipoprotein properties as those of TTP in normal tissues. Though slight proteolytic activity and fibrinolytic activity were demonstrated in the homogenate of gastric cancer tissues, it was noted that the TTP activity was different from these two activities by partial purification of TTP from gastric cancer tissues. The TTP activity of 9 homogenates of gastric cancer tissues was 301 +/- 289 (mean +/- SD) units per mg protein, being higher in homogenates of mucinous adenocarcinoma and signet-ring cell carcinoma than in those of tubular and poorly differentiated adenocarcinoma. The mean TTP activity of leukocyte homogenates from 14 patients with APL and one out of 4 patients with CML in blastic crisis was 81 +/- 76 units/10(7) cells. The TTP activity of the homogenates of leukocytes from 7 out of 18 patients with AML and another patient with CML in blastic crisis ranged from one to six units/10(7) cells with a mean of 3.3 +/- 1.2. The TTP activity of leukocyte homogenates from the other 11 cases of AML, two cases of CML in blastic crisis, 6 cases of CML, and one case each of ALL and CLL were less than one unit/10(7) cells. In leukemic patients, all cases with a value of more than 202 for the product of units of TTP activity per 10(7) cells and differential count (%) of leukemic cells in the bone marrow smear (MU value) were accompanied by DIC. The MU value of leukemic patients correlated well to the plasma fibrinogen and serum FDP levels. All patients with a MU value of more than 277 died of DIC when a sufficient amount of heparin was not administered. On the other hand, no DIC developed in any of the patients with a MU value of less than 90.(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: Blood Coagulation; Disseminated Intravascular Coagulation; Gastric Mucosa; Humans; Leukemia; Leukocytes; Stomach Neoplasms; Thromboplastin

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

Lysates of leukocytes of healthy persons and of patients with acute and chronic leukemias possess a weak tissue factor activity. This procoagulant activity is increased greatly when leukocytes are stimulated by endotoxin. The tissue factor is derived almost exclusively from the monocytes and not from lymphocytes and granulocytes. Monocytes are stimulated to the same extent by adherence to plastic surfaces. The fibrinolytic activity of lysates of mixed leukocytes is due to a nonspecific protease and a plasminogen activator. Only granulocytes can cause fibrinolyses. Endotoxin stimulation enhances the plasminogen activator but not the protease. Normal leukocytes and leukocytes of patients with chronic leukemias also exert an antithrombin activity.

Topics: Blood Coagulation; Disseminated Intravascular Coagulation; Endotoxins; Fibrinolysis; Humans; Leukemia; Leukocytes; Monocytes; Thrombin; Thromboplastin

Platelets were studied in a group of 10 patients with typical clinical course, morphological findings, and specific histochemical criteria for leukemic reticuloendotheliosis. In 8 of these, marked qualitative abnormalities were found. These included lack of aggregation following epinephrine stimulation (6 patients), and decreased platelet factor 3 availability following ADP stimulation (4 patients). In addition, platelets in 4 of the 10 patients were studied by electron microscopy. All had granular abnormality, and 1 case showed the presence of rough-surfaced endoplasmic reticulum. The functional and ultrastructural abnormalities of platelets reported here may be responsible for the clinically important bleeding episodes which were not attributable to thrombocytopenia in 2 of our patients. The findings also provide a clue to the basic nature of this histogenetically controversial malignancy.

Topics: Adenosine Diphosphate; Adult; Aged; Blood Platelets; Endoplasmic Reticulum; Epinephrine; Female; Humans; Leukemia; Lymphatic Diseases; Male; Middle Aged; Platelet Aggregation; Thromboplastin

Topics: Animals; Blood Coagulation Disorders; Blood Coagulation Tests; Blood Platelets; Disseminated Intravascular Coagulation; Embolism; Female; Humans; Leukemia; Male; Postoperative Complications; Pregnancy; Pregnancy Complications, Hematologic; Shock; Thromboplastin; Thrombosis; Wounds and Injuries

Topics: Acute Disease; Adolescent; Child; Child, Preschool; Humans; Leukemia; Leukemia, Lymphoid; Leukemia, Myeloid, Acute; Lymphatic Diseases; Thromboplastin

Topics: Blood Coagulation; Disseminated Intravascular Coagulation; Fibrinolysis; Humans; In Vitro Techniques; Leukemia; Leukemia, Myeloid, Acute; Leukocytes; Phosphatidylethanolamines; Thromboplastin

Topics: Bile Acids and Salts; Blood Coagulation Tests; Chromatography; Dextrans; Eosinophils; Humans; Leukemia; Male; Spectrophotometry; Thrombin; Thromboplastin

Topics: Chromatography, DEAE-Cellulose; Eosinophils; Humans; Leukemia; Male; Thromboplastin

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

Topics: Anemia; Anemia, Hemolytic; Anemia, Hypochromic; Anemia, Pernicious; Blood Coagulation; Blood Proteins; Chloramphenicol; Epidemiology; Epoetin Alfa; Erythropoietin; Haptoglobins; Hematologic Diseases; Hematology; Hemochromatosis; Humans; Iron-Dextran Complex; Leukemia; Polycythemia; Thromboplastin; Vitamin B 12

Topics: Blood Coagulation Tests; Hematologic Diseases; Hemorrhage; Humans; Leukemia; Thrombocytopenia; Thromboplastin

Topics: Adrenal Cortex Hormones; Blood Chemical Analysis; Child; Factor IX; Hemophilia A; Hemophilia B; Hepatitis; Hepatitis A; Humans; Infant; Infant, Newborn; Infant, Premature; Jaundice; Jaundice, Obstructive; Leukemia; Pharmacology; Physiology; Purpura; Thromboplastin

Topics: Blood Coagulation Tests; Genetic Diseases, X-Linked; Hemorrhagic Disorders; Humans; Leukemia; Leukemia, Hairy Cell; Lymphatic Diseases; Metabolism; Severe Combined Immunodeficiency; Thrombocytopenia; Thromboplastin

Topics: Annexin A5; Health Services; Humans; Leukemia; Thromboplastin

Topics: Acute Disease; Blood Platelets; Child; Humans; Infant; Leukemia; Thromboplastin

Topics: Humans; Leukemia; Leukocytes; Thromboplastin