Purified natural human fibroblast-derived interleukin-6 (nhlL-6) was obtained from a culture of a normal human fibroblast cell strain, and its effect on the platelet number in normal mice was examined. The platelet number in female BDF1 mice given subcutaneously 10, 50, or 250μg/kg of nhlL-6 for 5 consecutive days increased significantly in a dose-dependent manner, and the respective platelet counts were 131.4±3.7×104 (119%), 139.8±3.2×104 (127%), and 159.4±4.2×104/μl (144%). The count in control mice was 110.4±2.6×104/μl (100%). The significant increase in megakaryocyte number in the spleens of the mice receiving 250μg/kg was also observed. On the other hand, a substantial decrease in total bile acid in plasma and a remarkable increase in plasma immunosuppressive acidic protein (IAP), an acute phase protein, were found, and moderate but significant changes in parameters such as hemoglobin, hematocrit, plasma amylase, plasma alkaline phosphatase, plasma total cholesterol, and plasma free cholesterol were also observed.
Plasma levels of prothrombin fragment F1+2 were measured by a newly developed enzyme-linked immunosorbent assay (ELISA) in 44 healthy subjects, 34 patients with disseminated intravascular coagulation and 22 patients under stabilized warfarin therapy (INR>1.8). In this assay, rabbit antibody against F1+2 was used as a catching antibody, and rabbit antiprothrombin antibody as a tagging antibody. The mean plasma level of F1+2 in healthy subjects was 0.60±0.15 (SD) nM with a 95% range of 0.30-0.90nM. Plasma F1+2 was markedly elevated in patients with disseminated intravascular coagulation (6.02±2.90nM). In warfarin-treated patients, F1+2 concentrations (0.21±0.01nM) were significantly lower than those in healthy subjects. These results indicate that the measurement of F1+2 in plasma would be a sensitive and useful parameter for detection of hypercoagulable as well as hypocoagulable states in various diseases.
In 3 patients with hemophagocytic histiocytosis (HH) accompanied by disseminated intravascular coagulation syndrome (DIC) and multiple organ failure (MOF), a markedly significant decrease of plasma free protein S (PS) levels was found with markedly raised plasma C4b-binding protein (C4bp) levels as compared to non-HH DIC patients with MOF. Moreover, a significant elevation of serum cytokines, interleukin-lβ and tumor necrosis factorα, was observed when compared to non-HH DIC patients with MOF. Plasma free PS was inversely correlated with these two cytokines, while plasma C4bp was positively correlated with them. These results imply that abnormal plasma PS distribution, that is, markedly reduced free PS with raised C4bp-PS complex which was closely associated with hypercytokinemia would contribute to the pathogenesis of DIC and/or MOF in HH.
We analyzed and determined immunoglobulin class and epitope mapping of F VIII inhibitor antibodies developed in two nonhemophilic patients. Case 1 (H. N.) is a 58-year-old female with Diabetes Mellitus but no previous history of bleeding, and Case 2 (M. U.) is 29-year-old woman who developed an antibody to factor VIII in the postpartum period. The type of antibody derived from H. N. was IgG heavy chain, both κ and λ light chain, and type of antibody from M. U. was IgG heavy chain, κ light chain. The inhibitor disappeared after 14 month immunosuppressive therapy in the patient H. N., whereas only 2 months in the latter patient. Although the clinical course, titer of inhibitors, type of immunoglobulin light chain and underlying diseases were different, these two IgG components similarly reacted with the 92-kd and 80-kd polypeptides of plasma-derived F VIII and 44-kd and 72-kd polypeptides of thrombin treated F VIII, respectively. Control hemophilic antibody also bound to the same polypeptides. We concluded that inhibitors analyzed were not monoclonal and that the immunoglobulin class of the inhibitors and F VIII peptide reactivity were not necessarily correlated.
The efficacy, safety and usefulness of a highly purified concentrate of porcine factor VIII (MTI-9002; Hyate: C) was evaluated in 30 treatment episodes with 12 hemophilia A patients having inhibitor to human factor VIII in 11 institutes in Japan. A total of 44 infusions was given. Results were as follows. 1. The hemostatic efficacy was evaluated at 8 hours and 24 hours after the first infusion in each treatment episode. In 22 of 30 episodes (73.3%), excellent to good responses were obtained at both times. General hemostatic efficacy of each episode was also evaluated. In 26 of 30 episodes (86.7%), excellent to good responses were obtained. 2. When more units were infused than the calculated neutralizing dose based on the titer of porcine factor VIII inhibitor at the time of pre-infusion, an adequate rise in factor VIII: C was almost always seen after infusion of MTI-9002. However, even if factor VIII: C level remained low, desired hemostasis was occasionally obtained. 3. There were 8 infusions for 8 treatment episodes in 5 patients where a definite relationship between MTI-9002 and adverse reactions could not be eliminated. Since those patients treated with hydrocortisone responded well soon, it seemed that adverse reactions could be prevented and treated with hydrocortisone. 4. For patients who have difficulty controlling bleeding and low inhibitor cross-reactivity, it was felt that MTI-9002 was extremely useful.
Hypercholesterolemia is associated with an increased incidence of vascular complications. In order to assess the actual degree of activation of the coagulation and fibrinolytic systems in hypercholesterolemia, plasma levels of molecular markers were mesured in 43 patients with hypercholesterolemia. Mean plasma levels thrombin-antithrombin III complex (TAT), fibrinopeptide A (FPA), and plasminogen activator inhibitor (PAT-1) were significantly elevated in hypercholestrolemia as compared with healthy subjects. These findings indicated hypercoagulable state in hypercholesterolemia. Furthermore, we investigated the effects of pravastatin on plasma lipid, lipoprotein (a) and molecular markers of coagulation system. Pravastatin (10mg/day) was added to 22 patients with hypercholesterolemia whose plasma concentration of total cholesterol was more than 300mg/dl. The mean concentration of total cholesterol, triglyceride and phospholipid were significantly decreased after treatment. Lipoprotein (a) was not decreased. On the other hand, TAT, FPA, and PAT-1 were significantly decreased. This study suggested that pravastatin may have a preventive effect for the hypercoagulable state by improving hypercholesterolemia.
We measured plasma level of tissue type plasminogen activator (t-PA) antigen in patients with disseminated intravascular coagulation (DIC). Plasma levels of t-PA, PA inhibitor-1 (PAT-1), t-PA-PAI-1 complex, von Willebrand factor, thrombin-antithrombin III Complex (TAT), plasmin-α2 plasmin inhibitor complex (PIC), and FDP-D-dimer were markedly increased at the time of the onset of DIC but decreased after treatment. In most patients without leukemia, as the DIC score increased, plasma t-PA increased to 20 ng/ml or more. Plasma t-PA level before treatment for DIC was slightly higher in those showing poor outcome than it was in those showing good outcome. After treatment, t-PA level increased slightly in those showing poor outcome but decreased significantly in those showing good outcome. Plasma t-PA level was significantly higher in patients with organ failure than in those without. Positive correlation of plasma t-PA antigen level to PAI-1
The levels of automatically measured D-dimer in patients with various diseases were compared with those of fibrin degradation products (FDP). This assay was based on the absorbance changes in aggregations between D-dimer and latex granules coated with monoclonal antibodies against D-dimer. Patients with thrombotic diseases, malignancies, disseminated intravascular coagulation (DIC) and so on showed high levels of D-dimer measured by this assay. Furthermore, the levels of D-dimer were changed in correlation with fibrinopeptide A, the thrombin antithrombin III complex and the plasmin α2-plasmin inhibitor complex during treatment for DIC. The correlation cofficient was 0.768 between the levels of D-dimer and FDP in 572 samples (p<0.001) (Fig. 1). In 337 samples that showed levels of FDP exceeding 10μg/ml, 39 samples in 33 cases showed a D-dimer/FDP proportion of less than 0.15. In these patients, it was considered that the high levels of FDP were the result of increasing fibrinolysis activation more than coagulation (Table 1) or of fibrinogenolysis (Table 2).
We describe a previously unreported defect of protein S (PS) in proband (34-year-old, male) and his farther, characterized by low levels of cofactor activity for activated protein C(53-56%) contrasting with normal levels of total (101.0-108.7%) and free PS antigen (100.0-103.0%). The distribution of PS between the free form and the form complexed with the complement component C4b-binding protein (C4bp) was normal on crossed immunoelectrophoresis. The proband and his farther also have heterogeneous protein C (PC) deficiency, suffering from deep vein thrombosis (DVT) and pulmonary thromboembolism (PTE). His defects are transmitted in autosomal dominant fashion from his father. Other relatives carrying protein C dificiency (paternal aunt, paternal cousin, sister and nephew) were asymptomatic. We administrated 400mg of Bonzol® for two weeks to his father and investigated serial changes of PC, total PS, free PS, PS activity and C4bp. Laboratory studies showed that PC, total PS, free PS were increased, but C4bp and the ratio (free PS/PS activity) were not changed. These findings suggest that this case is probably a new type defect of protein S and the recurrent thrombotic disease in this family is due to inherited deficiency of protein C and dysfunctional protein S.
Klippel-Trénaunay-Weber syndrome is characterized by three major abnormalities; hemangioma of the nevus flammeus type, varicose veins, and hypertrophy of soft tissue and bone. These abnormalities usually affect a single extremity. In this paper, we describe a 23-year-old man with this syndrome complicated by disseminated intravascular coagulation (DIC) which was triggered or exacerbated by fracture of the right lower leg. Large ecchymosis from intramuscular injection sites, bleeding tendency from a puncture site of the right femoral artery, and a positive test for urinal occult blood were noted. Femoral arteriograph revealed many hemangiomas in the muscles surrounding fractured bones. Laboratory studies showed that prothrombin time was 13.0s (control 10.6s), fibrinogen 25mg/dl, antithrombin III 101%, FDP 95μg/ml, D-D dimer 33μg/ml, plasmin-α2 plasmin inhibitor complex (PIC) 14μg/ml, thrombin-antithrombin III complex 60.0ng/ml, and platelet counts 13.6×104/μl. These findings are compatible with DIC. Severe hypofibrinogenemia without thrombocytopenia suggested that primary fibrinolytic state may also coexist with DIC. Tissue damages of hemangioma associated with bone fracture may release activators of both coagulation and fibrinolytis, resulting in DIC and primary hyperfibrinolysis. Continuous infusion of heparin and replacement therapy with fresh frozen plasma (FFP) were started. With these treatments, DIC and primary hyperfibrinolytic state were controlled to the extent that the patient did not have obvious bleeding tendencies, even without amputation of the involved limb.
A Japanese family with familial elevation of plasma histidine-rich glycoprotein (HRG) is described. The propositus was a 41-year-old man, who complained of left hemiparesis and dysarthria. He was made a diagnosis of pontine and cerebellar infarctions by brain CT, brain MRI and cerebral angiography. His parents were not consanguineous, and none of his relatives suffered from thrombosis. His plasma HRG antigen measured by Laurell's method was 190% (normal range: 58-161%). No other hemostatic abnormalities were found. Family studies revealed the elevation of plasma HRG antigen in 10 family members. None of them had rheumatic heart disease or Behcet's disease, in which elevation of plasma HRG antigen was reported. These data suggest that familial elevation of plasma HRG was inherited in this family.
One autopsy case of a 69-year-old male with severe hemophilia A was reported, who died of a massive cerebellar hematoma. Normal follicles of lymphnodes and spleen were destructed and mesenteric lymphnodes were infiltrated by non-Hodgikin's lymphoma cells. Moreover multiple lesions of Aspergillus were also found. Since one year before his death, the number of CD4 lymphocytes had been below 200/mm3 and the levels of IgG and IgM had increased. Although he had remained as an asymptomatic carrier of HIV infection (CDC group II), he had some symptoms of AIDS from the pathological point of view. These findings indicated that anti-viral, anti-fungal and immunological therapies should be started before the decline of cellular immunity in the patients with HIV infection.