PubMed Health⌕ Search

Biomedical subjects

J D Ogle

Publications and source records attributed to J D Ogle.

At least 37 records · Page 2Linked to original sources
PubMed1991

The effects of in vivo administration of endotoxin on the functions and interaction of hepatocytes and Kupffer cells.

It was the purpose of this study to determine the effects of the in vivo administration of endotoxin on certain in vitro hepatocyte and Kupffer cell functions. An Alzet osmotic pump that contained endotoxin (LPS, 2.5 mg/100g) was implanted into the peritoneal cavity of 300g guinea pigs and delivered the endotoxin over a period of four days. In vivo administration of LPS did not cause a change in the in vitro release of albumin by isolated hepatocytes. However, when hepatocytes were co-cultured with Kupffer cells there was a significant decrease in albumin release for both control and LPS-treated animals. There was no difference between control and LPS-treated animals in the release of C3 by hepatocytes. However, there was a significant increase over the control group in C3 release by Kupffer cells from LPS-treated animals. When hepatocytes and Kupffer cells were cultured together, their release of C3 was not additive. Kupffer cells from LPS-treated animals released significantly greater amounts of PGE2 than control animals when stimulated in vitro with LPS. Thus, these Kupffer cells appeared to be primed to respond to an in vitro challenge of LPS. Kupffer cells from LPS-treated animals had significantly depressed antibody dependent cellular cytotoxicity (ADCC). This endotoxin model is useful for determining the in vivo effects of endotoxin on cellular function and gives some indirect evidence for the detrimental effects of LPS on the immune system and host defense.

Animals↗
PubMed1990

Effects of chemotactic peptide f-Met-Leu-Phe (FMLP) on C3b receptor (CR1) expression and phagocytosis of microspheres by human neutrophils.

FMLP caused maximal upregulation of CR1 on neutrophils at a concentration of 10(-8) M but caused maximal enhancement of CR1-dependent phagocytosis of C3b.IgG-coated microspheres only at a concentration of 10(-6) M. There were positive correlations between FMLP-mediated upregulation of CR1 and FMLP-mediated enhancement of phagocytosis (correlation coefficient = 0.73, slope = 2.2) and between FMLP-mediated upregulation of CR1 and FMLP-mediated increase in total cell-associated microspheres (correlation coefficient = 0.88, slope = 1.3). The phagocytic capacity of both untreated and 10(-6) M FMLP-treated neutrophils was completely inhibited by fluid phase C3b and partially inhibited by aggregated IgG. The data suggest that CR1 upregulation is required but is not sufficient for maximal phagocytosis by the leukocytes. The data also suggest that FMLP at the higher concentrations may impart a phagocytic function to CR1, activate other phagocytic receptors, elicit phagocytosis-inducing mediators or may elicit a separate mechanism of phagocytosis. During the study, it was observed that there was considerable individual variation among different neutrophil preparations with respect to CR1 expression and binding and phagocytic capacity.

Humans↗
PubMed1990

Comparison of abilities of recombinant interleukin-1 alpha and -beta and noninflammatory IL-1 beta fragment 163-171 to upregulate C3b receptors (CR1) on human neutrophils and to enhance their phagocytic capacity.

Both recombinant IL-1 alpha and -beta caused an upregulation of C3b receptors (CR1) on human neutrophils and caused a receptor-mediated enhancement of phagocytosis of C3b.IgG-coated microspheres by these leukocytes. The alpha and beta forms of the recombinant cytokine were of comparable potency regarding CR1 upregulation, although both generally had less than 25% of the potency of FMLP in this respect. Recombinant IL-1 beta was slightly more potent than the alpha form of the cytokine regarding phagocytosis of opsonized microspheres and, again, both forms were less potent than FMLP in causing an enhancement of phagocytosis by neutrophils. The synthetic noninflammatory immunostimulatory nonapeptide corresponding to residues 163-171 of IL-1 beta was completely inert with respect to upregulation of CR1 on neutrophils and the enhancement of phagocytosis by these cells. Thus this domain in the intact IL-1 beta molecule apparently is not involved in CR1 upregulation and the ensuing enhancement in phagocytosis by neutrophils, although it is apparently important in the immunostimulatory activity regarding the proliferation of lymphocytes.

Antigens, CD↗
PubMed1990

The effects of cytokines, platelet activating factor, and arachidonate metabolites on C3b receptor (CR1, CD35) expression and phagocytosis by neutrophils.

The cytokines tumor necrosis factor alpha (TNF alpha), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), and interleukin 1 (IL 1) all caused an upregulation of C3b receptors (CR1) on neutrophils that ranged from around 76% (G-CSF and IL 1) to 93% (TNF alpha and GM-CSF) of the upregulation obtained by pretreatment of the neutrophils with the chemotactic peptide FMLP. However, only TNF alpha and G-CSF caused a significant increase in phagocytosis of opsonized microspheres. Platelet derived growth factor, interleukin 2, and transforming growth factor beta had no effect on either of these parameters. The mediators platelet activating factor (PAF) and leukotriene B4 (LTB4) both caused a large upregulation of CR1 (93% and 80%, respectively, of the FMLP-mediated value); however, only PAF caused a significant enhancement of phagocytosis by the neutrophils. Prostaglandin E2 and thromboxane B2 had no effect on these parameters. Considerable individual variation was observed among some of the untreated and mediator-treated neutrophil preparations regarding CR1 expression and phagocytosis. The upregulation of CR1 and associated increase in phagocytic capacity of neutrophils caused by certain cytokines and other mediators may be important in host defense. Also the lack of enhancement of phagocytosis accompanying an upregulation of CR1 is unusual and may have important implications regarding the cellular mechanisms of phagocytosis by neutrophils.

Antigens, CD↗
PubMed1989

Effect of antibiotics on CR1 receptor levels of human neutrophils and on the binding and phagocytosis of opsonized polystyrene microspheres by these leucocytes.

Fourteen antibiotics were studied for their effects on the following properties or functions of normal, human neutrophils: (1) the expression of CR1 receptors; (2) the total binding of C3b.IgG-coated polystyrene microspheres in the presence of cytochalasin D to inhibit phagocytosis; (3) the net phagocytosis of the opsonized microspheres; (4) the residual, external binding after phagocytosis. Fluorescence, flow cytometric methods were used to determine binding and phagocytosis of the model target particles. Only nafcillin, a penicillin, caused a decrease (33 per cent) in phagocytic capacity of the neutrophils at a physiologically significant dose (serum level); the antifungal antibiotic, amphotericin B, caused a 30 per cent increase in phagocytic capacity. These small changes in neutrophil phagocytic capacity may not be physiologically significant. There were no significant differences in the four measured parameters caused by other antibiotics tested at physiologically significant doses.

Amphotericin B↗
PubMed1989

Production and release of C3 by cultured monocytes/macrophages isolated from burned, trauma, and septic patients.

The release of C3 under various conditions by cultured monocytes/macrophages isolated from burned, trauma, and septic trauma patients was determined. When monocytes were cultured for up to 14 days there was no difference in production and release of C3 by normals and patients; C3 production rose slowly at first then rose rapidly from 8-14 days of culture. C3 production continued for up to 28 days of culture. When lymphocytes were present with the monocytes for the first 24 hr of culture there were differences in C3 production by normal and patient cells at various times of culture. There were differences in C3 release by patients' 24 hour-cultured cells at certain postburn and injury days. Macrophages from burned patients released less C3 than normal cells and macrophages from septic trauma patients released more C3 than normal cells.

Burns↗
PubMed1989

The immunosuppressive effects of the in vivo administration of endotoxin as influenced by macrophages.

It is well documented that endotoxin can have immunosuppressive effects on lymphocytes and induce the production and secretion of monokines which act on the lymphocytes. To delineate the interaction between macrophages and lymphocytes more clearly, 0.15 mg of lipopolysaccharide (LPS) (E. coli 0111:B4) was injected into Hartley guinea pigs intraperitoneally twice a day for 7 days (saline for control group). Seven days after the last injection, spleens were taken and lymphocyte proliferation was determined in the presence and absence of macrophages. When macrophages were present, there was a significant suppression of lymphocyte proliferation when PHA and PWM were used as mitogens. There was no suppression of proliferation when the macrophages were removed. Splenic macrophages were also cultured in the presence and absence of LPS and their supernatants analyzed for PGE2 and TXB2. There was no significant difference between the endotoxin and control groups for PGE2 or TXB2 production in the presence and absence of LPS. However, the endotoxin group had significant decreases in serum levels of C3 postinjection of endotoxin which could indicate C3 degradation by LPS. Taken together these results give further evidence that macrophage products in addition to PGE2 can inhibit lymphocyte proliferation. C3 degradation products could possibly stimulate macrophages to produce inhibitors of lymphocyte proliferation or induce suppressor cells.

Animals↗
PubMed1988

Phagocytosis of opsonized fluorescent microspheres by human neutrophils. A two-color flow cytometric method for the determination of attachment and ingestion.

A highly reproducible two-color fluorescence cytometric method is described for determining the amount of receptor-mediated and of non-specific phagocytosis by human neutrophils of polystyrene microspheres that have been covalently coated with C3b, iC3b, IgG, mixtures of these, BSA or human F(ab')2. The method includes a correction for externally bound particles and thus measures net phagocytosis. The method involves the incubation of neutrophils with coated green fluorescent microspheres in buffer alone or with cytochalasin D to inhibit phagocytosis followed by staining the externally bound microspheres with red fluorescent antibodies to the immobilized ligand, and determining the green and red fluorescence in a dual laser fluorescence activated flow cytometer. The red to green fluorescence ratio of the mixtures containing cytochalasin D allows for a correction of the green fluorescence due to externally bound microspheres in the mixtures not containing cytochalasin D. The corrected green intensities thus represent net phagocytosis. The specificity of receptor-mediated phagocytosis was confirmed by inhibition with fluid-phase C3b or iC3b or monoclonal antibodies to the receptors. The method can be applied to the determination of both adherent and suspension phagocytosis and can be used as a general model of the phagocytosis of bacteria by neutrophils.

Buffers↗
PubMed1988

The production of C3, PGE2 and TxB2 by splenic, alveolar, and peritoneal guinea pig macrophages.

The synthesis and release of C3, PGE2, and TxB2 by cultured splenic, alveolar and peritoneal guinea pig macrophages in 24 hour culture was determined. There were significant differences in C3 production among all three sources of macrophages. Splenic macrophages produced significantly less PGE2 than alveolar or peritoneal macrophages. Peritoneal macrophages produced significantly more TxB2 than splenic or alveolar macrophages. The cells from the different sources appear to be different populations of macrophages.

Animals↗
PubMed1987

Inhibition of lymphocyte proliferation by fluid-phase C3b as influenced by macrophages.

The effect of fluid-phase C3b on mitogen-induced lymphocyte proliferation in the presence and absence of macrophages was studied. In general, C3b inhibited the proliferation of lymphocytes when monocytes (macrophages) were present. The degree of inhibition by C3b was different for B and T lymphocytes and varied for different subpopulations of lymphocyte classes. In the absence of monocytes (macrophages), there was insignificant inhibition by C3b of lymphocyte proliferation, and thus the observed inhibition appeared to be due to the effect of C3b on the monocytes/macrophages present in the mixed lymphocyte preparations.

Cell Division↗
PubMed1986

Fibronectin coating of an experimental PTFE vascular prosthesis.

Because of the known inefficiency of endothelial cell seeding of vascular prostheses, some investigators have pretreated their grafts with fibronectin in an effort to increase endothelial cell adhesion. However, the most efficient method of coating vascular prostheses with fibronectin has not yet been determined and more importantly, the durability of this coating following restoration of pulsatile perfusion has not yet been established. This experiment measured the adsorption of fibronectin which was perfused at three different concentrations through an experimental, highly porous PTFE vascular graft. Most efficient coating of the prosthetic appeared to occur with a 250 mg/ml concentration of fibronectin perfused through the graft for 60 min. Grafts so prepared were subsequently studied in a pulsatile perfusion system which mimicked the canine circulation. Fifteen additional fibronectin-coated grafts were also interposed in the canine carotid artery. Flow was restored, grafts were removed at varying intervals, and fibronectin disappearance was measured. In both the in vitro and in vivo models, three distinct phases of fibronectin dissociation were seen: an initial rapid loss which resulted in an average 30.5% retention of the initial concentration of fibronectin following 30 min of perfusion; a second phase, which occurred from 30 to 120 min, demonstrated an average 2.5% loss of fibronectin per hour; and a final phase, from 2 to 24 hr, during which no significant additional loss of fibronectin was seen. This study describes a new method by which PTFE vascular prostheses can be effectively coated by perfusing them with physiologic concentrations of fibronectin and demonstrates that the resulting bond between fibronectin and prosthetic is stable following restoration of pulsatile flow.

Adsorption↗
PubMed1985

Studies on the binding of C3b-coated microspheres to human neutrophils.

A method is described for the quantitation of C3b receptors on human neutrophils using a mixture of C3b-coated fluorescent and C3b-coated non-fluorescent microspheres. The method measures the "sterically available' C3b receptors on the cells, for example, the receptors available to opsonized bacteria. The use of mixtures of fluorescent and non-fluorescent microspheres resulted in lowered fluorescence intensities of the microsphere-coated neutrophils that were well within the fluorescence limitations of fluorescence activated cell analyzers or sorters used in the assay procedure. These mixtures also allowed the distribution of the C3b-coated microspheres around the neutrophils to be easily visualized in the fluorescence microscope. The binding of the C3b-coated microspheres to the neutrophils was shown to be receptor mediated by typical saturable binding kinetics, by complete inhibition by fluid phase C3b, but not by other proteins and by nearly complete inhibition by anti-C3b receptor antibody. Several parameters that could affect the binding of C3b-coated microspheres to neutrophils were studied; these included time and temperature of incubation of the microspheres with the cells, the diameter of the microspheres, the C3b content of the C3b-coated microspheres, the presence of metal ions, azide, EDTA, protein (BSA, IgG), soybean trypsin inhibitor in the buffers, and the method of isolation of the neutrophils. The C3b-coated microspheres were evenly distributed around the neutrophils in almost all of the cases; however, the neutrophils used in these studies were not activated and were not phagocytosing. The method is extremely reproducible and sensitive in detecting small changes in number of C3b receptors on cells.

Complement C3b↗
PubMed1985

Determination of C3b receptors on normal and patient polymorphonuclear neutrophils with C3b-coated fluorescent microspheres.

A method, devised in the authors' laboratories, for the determination of C3b receptors on normal and patient neutrophils using C3b-coated fluorescent microspheres, was applied to the quantitation of C3b receptors on the neutrophils of several patients suffering from burns and trauma and a patient with pancreatitis. From three to 11 days in the clinical course the relative number of C3b receptors was, or rose to, two to ten times the number of receptors present at later times in the clinical course and, in most of the cases studied, the increase in C3b receptor number coincided with enhanced neutrophil bactericidal function. The rise in C3b receptor number was ascribed to up-regulation by C3a and C5a des Arg from complement activation and also, in the cases where sepsis occurred, to the presence of bacterial chemotactic peptides. Preliminary experiments with zymosan-activated serum and the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine confirmed this explanation.

Blood Bactericidal Activity↗
PubMed1984

Inhibition of bacterial clearance in the guinea pig by fluid-phase C3b.

The large-molecular-weight activation product C3b of C3, the third component of complement, loses its opsonic properties when it is free in solution (fluid phase) rather than attached to a particle, eg, a bacterial cell. Human fluid-phase C3b, which has been shown in vitro to be an inhibitor of the bactericidal function of human neutrophils, was injected intradermally or intraperitoneally into guinea pigs along with various species of bacteria, and the clearance rate of the bacteria was measured. The injected C3b caused a large increase in the time required for the clearance of the bacteria. The inhibition by C3b of clearance of the injected bacteria was dependent on the bacterial species; bacteria that depended strongly on complement for opsonophagocytosis was cleared at a much lower rate in the presence of fluid-phase C3b than were bacteria that did not have the corresponding dependence on complement.

Animals↗
PubMed1984

Complement-induced expression of cryptic receptors on the neutrophil surface: a mechanism for regulation of acute inflammation in trauma.

We explored the hypothesis that identified changes in neutrophil function in patients with acute injury result from in vivo exposure to C5a. To evaluate this hypothesis, we performed a battery of tests on 26 trauma patients (14 with blunt injury, 12 with penetrating injury). Measured were plasma levels of the complement activation products C3a and C5a; neutrophil chemotaxis to C5a and N-formyl-methionyl-leucyl-phenylalanine (FMLP); neutrophil receptors for FMLP and C3b; and superoxide response to FMLP and serum-opsonized zymosan. Patient responses measured within 48 hours of admission were divided into two groups based on neutrophil migratory response to C5a. Patients unresponsive to C5a (but responsive to FMLP) showed elevated plasma C3a levels (248 +/- 6 ng/ml) compared with patients with normal C5a migratory response (104 +/- 8 ng/ml). FMLP receptor number was markedly increased in the chemotactically deactivated group (group I: 155,680 +/- 100; group II: 51,200 +/- 200) and receptor affinity was diminished. Binding activity of C3b increased in the C5a-unresponsive cells to 126% that of controls versus 94% for normally responsive patient cells. Superoxide production was found to be significantly increased in patient cells with increased receptor numbers. These results support the concept that a subgroup of trauma patients manifest plasma and neutrophil changes compatible with complement activation. The neutrophil changes identified demonstrate a state of cellular activation. The clinical significance of these results may reside in a risk of pulmonary microvascular injury if activated cells are marginated and then subsequently stimulated.

Chemotaxis, Leukocyte↗
PubMed1983

Inhibition of neutrophil function by fluid phase C3b of complement.

A high-molecular-weight fragment of C3 was isolated from normal human serum by column chromatography, was generated by incubation of serum at 37 degrees C with inulin, and was produced from highly purified C3 by limited digestion with trypsin. This product was shown to inhibit the antibacterial function of neutrophils by using Escherichia coli O75 as the main test organism. The inhibitor reacted with anti-C3b and anti-C3c, but not with anti-C3B (anti-native C3) or anti-C3a. The manner of preparation of the inhibitor, the sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern, and the amino acid composition of the inhibitor indicated that it was fluid phase C3b. The inhibitor of neutrophil function (fluid phase C3b) was shown to bind to C3b receptors or acceptors on sheep erythrocytes in a model system.

Amino Acids↗