Partial characterization and purification of a rabbit granulocyte factor that increases permeability of Escherichia coli (original) (raw)
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Cell Proliferation, 1973
The granulocytic chalone is secreted by mature granulocytes and inhibits 3H-thymidine incorporation of proliferating granulocytes in vitro. The effect and the cell line specificity of this chalone was assessed with the in vivo diffusion chamber culture technique. Tests were carried out on cultures from normal mouse bone marrow cells and mouse and rat blood leucocytes. The majority of the DNA synthesizing cells in marrow cultures were proliferating granulocytes. Macrophages and immunoblasts proliferated in rat leucocyte cultures, when the chambers had been carried for 5 days in host mice. Repeated chalone or control injections were given i.p. to the host mice during 6-7 hr prior to 3H-thymidine injection. Isotope uptake of proliferative granulocytes was reduced by the chalone treatment. No such effect was found on the rat immunoblasts and macrophages. The viability of cultured cells was apparently not affected by the chalone treatment.
Interference of granulocyte function by the vehicle of miconazole
Antimicrobial Agents and Chemotherapy, 1978
The vehicle of miconazole inhibited adherence and leukotaxis, but did not affect nitroblue tetrazolium reduction and trypan blue exclusion by granulocytes at 0.33% concentration and above. Information on the effects of miconazole, a new and effective antifungal drug, on host defense function is meager (3). Since it is agreed that intact leuokocyte function is important especially in the infected host, we studied the influence of miconazole and its liquid vehicle on granulocyte functions. Miconazole base dry powder (lot no. A 29/2) and its vehicle marked "placebo" (lot no. 618.095, containing 0.115 ml of polyethoxylated castor oil, 1.62 mg of methylparaben, 0.18 mg of prophyloparaben, and pyrogen-free water to make 1 ml) were kindly provided by Janssen Research and Development, Inc., New Brunswick, N.J. For our studies a stock solution was made by dissolving 41.6 mg of miconazole base powder in 10 ml of vehicle. Further dilutions of the stock solution and dilutions of the vehicle were made using medium 199. Granulocytes were prepared from heparinized venous blood by sedimentation of erythrocytes at 1 x g for 1 h and centrifugation of the leukocyte-rich plasma at 500 x g for 5 to 10 min. The cell pellet obtained was then washed once in medium 199 and adjusted to a concentration of 5 x 106 granulocytes per ml. Only medium 199 was used in all experiments. Granulocyte adherence was measured by the Hemnalini and Martin modification (Program Abstr. Intersci. Conf. Antimicrob. Agents Chemother. 15th, Washington, D.C., Abstr. no. 68, 1975) of the method described by MacGregor et al. (4), using 40 mg of scrubbed nylon fibers (in LP-1 Leuko-Pac leukocyte filter, Fenwal Laboratories, Morton Grove, Ill.) packed inside each plastic tuberculin syringe, with the length of the packed column adjusted to exactly 15 mm.
Granule release by polymorphonuclear leukocytes treated with the lonophore A23187
The Anatomical Record, 1977
Polymorphonuclear leukocytes (PMN's) incubated three to eight minutes a t 37OC in medium containing 1 x M of the ionophore antibiotic A23187 released their cytoplasmic granules into the extracellular medium. Transmission electron microscopy of treated cells showed microfilament bundles extending between adjacent granules within the cytoplasm and between granules and the plasma membrane. Tiny dense projections (beads) 8-12 nm in diameter were observed along segments of the cytoplasmic surface of the plasma membrane with a periodicity of 20-30 nm. These beads were observed on the plasma membrane only in the vicinity of intra-or extracytoplasmic granules. The structural relationships of the beads with the plasma membrane microfilaments suggest they play a role in the process of ionophore-induced granule release from polymorphonuclear leukocytes. The ionophore antibiotic A23187, extracted from cultures of Streptomyces chartreusensis, has been shown to alter the permeability of biological membranes to calcium (Reed and Lardy, '72). Such phenomena as lymphocyte mitogenesis (Hovi et al., '76; Luckasen et al., '74), salivary gland secretion (Prince et al., '731, leukocyte chemotaxis (Estensen et al., '76; Wilkinson '751, lymphocyte agglutination (Poste and Nicholson, '76) and capping (Poste and Nicholson, '76; Schreiner and Unanue, '76) have been shown to be initiated, or otherwise affected by the changes in intracellular calcium levels induced by this ionophore. Recent biochemical evidence has indicated that this ionophore enhanced secretion of the specific granule enzyme, lysozyme, from human neutrophils, thus implicating calcium in the induction of its release (Estensen et al., '76; Goldstein et al., '74). In the present report, transmission and scanning electron microscopy were employed to assess the structural changes that occur during granule release in rabbit PMN's exposed to the ionophore A23187.
Acta pathologica et microbiologica Scandinavica. Section B: Microbiology and immunology
Disintegration of rabbit polymorphonuclear leucocytes in suspension is achieved by extrusion under controlled pressure. The operating pressure is the key factor influencing the degree of disintegration. The number of cells disintegrated increases with higher pressure. At 35 kp/cm2, 80-90 per cent of the cells are disintegrated. The disintegrated material was subsequently analysed by following the subcellular distribution of leucocyte A and B granule marker enzymes. The analysis reveals that although 80-90 per cent of the cells are disintegrated, a high degree of subcellular integrity remains. Leucocyte A and B granules, abundantly present in the disintegrated material, were separated from soluble components by zonal centrifugation. Extrusion under controlled pressure as a cell disintegration method is discussed, particularly with reference to the study of polymorphonuclear leucocyte structure and function.
Haematologica, 2008
Granulocyte transfusion has been proposed as a bridging therapy for patients with prolonged periods of chemotherapy-induced neutropenia, who suffer from severe fungal and bacterial infections. To recover, adequate numbers of granulocytes are required when the patients are refractory to standard treatment. The aim of this study was to assess the functional characteristics and efficacy of granulocyte colony-stimulating factor/dexamethasone-mobilized granulocytes used for transfusions.
Virchows Archiv. B, Cell pathology including molecular pathology, 1982
The inhibitory effects of partly purified low molecular weight granulocyte extracts (GE) from human peripheral blood were investigated on mouse myelopoietic cells in agar cultures. By repeated freezing and thawing the inhibitory effect of the extract was lost, and instead a stimulatory effect was seen. The stimulation was seen mainly as an increase of colonies and clusters from the sixth day of culture. The degree of inhibition depended on the length of the preincubation time. After half an hour a stable and lasting inhibition was found. The biological effects of the extracts were temperature dependent. Inhibition or stimulation was seen when the target cells were exposed to GE at 37 degrees C and not at 4 degrees C. Colony formation in agar showed circannual variation with maxima in spring and autumn and minima in summer and winter. The data indicated that the inhibitory effect of GE did not show circannual variations.
Phagocytosis of bacterial aggregates by granulocytes
European Journal of Epidemiology, 1988
A study was conducted on the granulocytic phagocytosis of bacterial aggregates obtained under ideal environmental conditions. For the strains studied, aggregation was favored by low salt concentrations, low pH and temperatures between 30°C and 40°C. Our results show that the phagocytic capacity of granulocytes depends on the type and size of these aggregates. Those formed by a smaller number of cells are more easily phagocytized than the larger ones.
Evaluation of a short term in vitro test for granulocyte chalone activity
Virchows Archiv. B: Cell pathology, 1976
A short term in vitro test for granulocyte chalone activity eas examined for its specificity and reliability. The test used the inhibition, by granulocyte extracts, of 3H-thymidine (3H-Tdr) uptake in to the acid-insoluble material by rat bone marrow cells in vitro to measure possible chalone activity. Among the many possible 3H-Tdr artifacts pool size dilution by Tdr contained in the extracts was excluded using an E. coli mutant requiring thymine. Several amino acids and biogenic amines do not affect the test. However, continuous and pulse labelling of bone marrow cells with 3H-Tdr, viability tests and micro flow fluorometric measurements of the cell cycle distribution following colcemid treatment strongly suggests that the cells do not proliferate in vitro during short term incubation, since practically no cells enter the S-phase, cells in the S-phase die and few if any cells proceed through G2 and mitosis. Moreover, the test cannot exclude cytotoxicity. Thus, the in vitro test may...