Electron microscopic studies of circulating neutrophil granulocytes of a boy suffering from Fatal Granulomatous Disease (FGD) showed two abnormal kinds of cytoplasmic granules, both larger than normal granules. One was characterized by a homogeneous matrix of very low electron density, the other contained one or more crystalloid inclusions of larger size than found in normal neutrophils. In neutrophil granulocytes from adults with long-standing bacterial infection, by light microscopy showing toxic granulation, the granules displayed the same ultrastructural abnormalities as in FGD. It is concluded that the ultrastructure of the granules of neutrophils in FGD reflects non-specific toxic granulation. The functional defect of the granules in FGD has no known morphological counterpart.
Of 20 patients suffering from severe haemophilia-A, 4 were complicated by the development of inhibitor directed against factor VIII. Two of the 4 patients were siblings. Before detection of the inhibitor the patients were treated with cryoprecipitate, AHG (Cohn fraction I-O) and blood transfusions.
In vivo microscopic observations of splenic tissue transplanted into modified Algire back chambers in mice revealed increased vascularity of the transplant following injections of various doses of erythropoietin. Concomitant histologic and peripheral blood studies suggested that this was vasoproliferation that occurred independent of the process of erythropoiesis.
Five young, healthy women with regular menstruations and who were not taking oral contraceptives were followed with repeated blood samples for 5 weeks. On each individual, about 15 determinations were made of the number of platelets, the fibrinogen and plasminogen concentration, the P & P-test, the Factor VIII activity and the spontaneous fibrinolytic activity of the euglobulin fraction of plasma on fibrin plates. It was shown that the variation in these parameters in the individual was smaller than the variation between individuals and that distinct individual ranges could be found for some women. This was most pronounced with respect to the number of platelets, the fibrinogen concentration and the P&P-test. The importance of knowing normal intra-individual ranges of different parameters in blood coagulation and fibrinolysis is stressed. Cyclic variations in the number of platelets were found during the menstrual cycle but there were no such variations in the other parameters studied.
Intracellular lysozyme concentration was measured in neutrophilic granulocytes from 25 patients with multiple myeloma. At diagnosis intraneutrophil lysozyme activity was significantly reduced (mean reduction 50%). During clinical remission after 1-4 months of intensive chemotherapy values were normalized. In 18 cases studied at various stages of the disease from 6 to 70 months after diagnosis there was a significant negative correlation between the duration of the disease and neutrophil lysozyme concentration. The decrease in neutrophil lysozyme concentration was significantly correlated to clinical disease activity and the percentage of plasma cells in bone marrow aspirates, whereas there was no correlation between the concentration of M-protein in serum and the neutrophil lysozyme concentration. Plasma lysozyme concentration was normal. In contrast, neutrophil lysozyme concentration was normal in 18 patients with stage III-IV malignant lymphoma. Plasma lysozyme in this group was significantly higher than normal. The difference in neutrophil lysozyme patterns between multiple myeloma and malignant lymphoma supports the hypothesis that the defect in neutrophil maturation seen in malignant blood disorders is directly related to the infiltration of the bone marrow by pathologic cells.
The kinetics of simultaneously injected 111In- and 51Cr-labelled platelets have been assessed in 40 subjects, 13 of them thrombocytopenic. 4 platelet survival models were applied. The mean life-time (MLT) of 51Cr-platelets from non-thrombocytopenic individuals was found to be slightly, but significantly, longer than that of 111In-platelets by applying linear and exponential models for data fitting. The in vivo recovery (IVR) of 111In-platelets was significantly higher than that of 51Cr-platelets in this patient group when using all 4 models. In the group of thrombocytopenic patients no statistically significant differences in MLT or IVR were found between 111In- and 51Cr-platelets. However, for each of the 11 51Cr-labelled platelet suspensions with the shortest MLT, a longer MLT was observed in the corresponding 111In-platelets, a finding probably related to antibody-induced elution of 51Cr-activity. The same mechanism might be responsible for an increasing 111In-/51Cr-recovery ratio in the early post-injection period. The efficiency of platelet isolation from blood prior to labelling seemed to influence the IVR, inasmuch as the difference in IVR between 111In- and 51Cr-platelets was eliminated in the group where the yield of 111In-platelets surpassed that of the 51Cr-platelets by more than 15%.
Both polymorphonuclear (PMN) and mononuclear (MN) leucocytes constitute targets for glucocorticoid hormones. In order to comparatively characterize the earliest steps of steroid action in these cell populations, we investigated the concentrations and specificities of glucocorticoid receptors in purified human PMNs and MNs by a whole-cell binding assay using (3H)dexamethasone as the ligand. PMNs and MNs were found to contain the same amounts of glucocorticoid receptors (4720 and 4900 receptor sites/cell, respectively). The equilibrium dissociation constant (KD) of the interaction between the cellular receptor and (3H)dexamethasone was about the same (1 x 10(-8)M) in both cell populations. No significant difference in the specificities of the steroid binding sites in PMNs and MNs were found; competition studies revealed the following order of relative binding affinities for a number of compounds: betamethasone greater than dexamethasone greater than prednisolone greater than cortisol greater than deoxycorticosterone = progesterone greater than aldosterone greater than testosterone greater than estradiol-17 beta. We conclude that known differences in the sensitivities of PMNs and MNs to glucocorticoids are apparently not caused by differences in the concentrations or characteristics of their glucocorticoid receptors.
To investigate the source and the mechanisms of synergistically enhanced colony-stimulating activity elaboration by the coincubated monocyte-macrophages and T lymphocytes, we simultaneously prepared conditioned media both from the coincubated monocyte-macrophages and T lymphocytes (ratio 1:3) in the presence of phytohemagglutinin (1%) or methanol extraction residue of bacillus Calmette-Guerin (50 micrograms/ml) and from the isolated T lymphocytes that had been primed with monocyte-macrophages in the presence or absence of phytohemagglutinin or methanol extraction residue of bacillus Calmette-Guerin. Subsequently, colony-simulating activity in various conditioned media was assayed using light-density (less than 1.070 g/ml), nonadherent normal human marrow cells. Live monocyte-macrophages synergized with and significantly (P less than 0.01) agumented the T lymphocyte-derived colony-stimulating activity elaboration; while killed monocytes-macrophages had no such effect. Similarly, actinomycin D and cycloheximide not only diminished monocyte-macrophage colony-stimulating activity elaboration but also reduced their synergistic interaction with T lymphocytes and their ability to augment the T lymphocyte-derived colony-stimulating activity elaboration. In contrast, mitomycin C failed to diminish both - monocyte-macrophages' ability to synergise with T lymphocytes and also to augment T lymphocyte-derived colony-stimulating activity. These data suggest that monocyte-macrophages require an intact transcriptional and translational processes, but not DNA synthesis for synergising with T lymphocytes or for augmenting T lymphocyte colony-stimulating activity elaboration.
A case of acquired dyserythropoiesis with inter-erythroblastic connections is reported in a patient with chronic myeloid leukaemia, developing a terminal acute hepatic failure related to hepatocarcinoma. The erythroblastic series was abundant but only made of clusters grouping 10 to 20 closely adherent cells. The cellular membranes showed linear junctions or were interdigitated and the intercellular space was occupied with electron dense ferritin granules. This non specific aspect of dyserythropoiesis may be related to the hepatic carcinoma, which was probably induced by busulfan therapy.
Bleeding is a common complication in patients suffering from multiple myeloma. In some cases a defect in fibrin formation has been suggested as one possible cause of haemorrhagic tendency. As shown in this investigation the defect in fibrin formation, ascertained using PAGE, is due to a lack of alpha-chain polymerization of fibrin monomers in 5/11 patients with IgG myeloma and in 2/5 patients with IgM paraproteinaemia. No disturbed fibrin polymerization could be observed in IgA myeloma (n = 6). Factor XIII concentrations of subunit A and to a lesser extent of subunit S (Laurell technique) were highly elevated in all cases with regular fibrin formation. comparable values were obtained by measuring the transamidase activity of factor XIII by incorporation of 14C-labelled purtrescin into casein. Levels up to 600% of normal could be recorded. In contrast, all patients with a lack of alpha-chain polymerization had a factor XIII activity within the normal range. Addition of factor XIII concentrate to plasma from patients with defective fibrin formation led in 5/8 cases to a partial cross-linking of alpha-monomers. we conclude that in some cases paraproteins can inhibit the factor XIII and prevent its action on fibrin.
By infusion of large amounts of human fibrinogen into rabbits, the ethanol gelation test turned positive at various plasma concentrations of fibrinogen, depending on the fibrinogen preparation used, and on individual differences among the animals. After incubation of the fibrinogen preparation in the cold the precipitated material gave a positive ethanol gelation test at lower fibrinogen concentration whereas the ethanol gelation test remained negative in the supernatant at a higher fibrinogen level than that of the starting material. It is concluded that the finding of a positive ethanol gelation test is not due to the fibrinogen concentration per se. Most likely the test becomes positive due to the presence of soluble fibrin in the commercial preparations.
In a patient with eosinophilic leukaemia, serial chromosome studies using the Giemsa banding technique revealed a similar marker chromosome, identified as an isochromosome 17, in all bone marrow metaphases analysed. There was no Ph-1-chromosome. The cytogenetic data support the view that eosinophilic leukaemia is a specific disease entity, and suggest that both eosinophilic and neutrophilic cells are involved in the leukaemic process.
Neutropenia is an almost constant feature of Chediak-Higashi syndrome (CHS). There is evidence for a central mechanism of neutropenia. Ultrastructural studies of the bone marrow from a child with CHS showed marked autophagic phenomena within myeloid precursor cells and mature neutrophils. Autophagic vacuoles were randomly distributed in the cytoplasm of the cells from the granulocytic series and some of them contained giant granules which thus appeared particularly resistant to the autophagic process. The vital cellular damage through endophagocytosis suggests the possibility of intramedullary destruction as an explanation for neutropenia.
The immunologic consequences of replacement therapy have been studied in 2 pairs of young haemophilic monozygotic twins. In both pairs one cotwin only developed a factor VIII specific antibody. This non-concordance illustrates the influence upon a specific immune response of non-genetic factors since the blood products, administered to each brother-pair, were not at all time identical nor were the regimens of transfusion therapy.
A 23-year-old patient with Gaucher's disease and IgG kappa monoclonal gammopathy is described. The decrease of the serum immunoglobulin level after splenectomy and the immunoflourescence study of the spleen indicate a predominant splenic location of the secreting plasma cells.
