scholarly journals IgM anti-Fc gamma R autoantibodies trigger neutrophil degranulation.

1991 ◽  
Vol 173 (6) ◽  
pp. 1473-1482 ◽  
Author(s):  
P Boros ◽  
J A Odin ◽  
T Muryoi ◽  
S K Masur ◽  
C Bona ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Hydrolytic Enzymes ◽  
Phosphatidyl Inositol ◽  
Human Neutrophils ◽  
Enzyme Release ◽  
Fab Fragments ◽  
Transmission Electron ◽  
Neutrophil Degranulation ◽  
Inflammatory Processes

Anti-Fc gamma R IgM monoclonal antibodies (mAbs) isolated from lipopolysaccharide-stimulated spleen cells from tightskin (TSK) mice were found to be polyspecific, reacting with a wide variety of molecules, including double-stranded DNA, topoisomerase, RNA polymerase, and different collagen types. Approximately 60% of the polyspecific IgM mAbs have anti-Fc gamma R specificity. These anti-Fc gamma R mAbs induce the release of hydrolases from both azurophil and specific granules of human neutrophils. 25-45% of the total cellular content (determined in Nonidet P-40 lysates) of neutrophil elastase, 10-25% of beta-glucuronidase, and 30-50% of alkaline phosphatase was released after incubation with the mAbs. The degranulation process was accompanied by dramatic morphological changes shown by scanning and transmission electron microscopy. The release of hydrolytic enzymes stimulated by the IgM anti-Fc gamma R mAbs was inhibited by preincubation of neutrophils with Fab fragments of either anti-human Fc gamma RII (IV.3) or anti-human Fc gamma RIII (3G8) mAbs. The binding of the anti-Fc gamma R TSK mAbs to human neutrophils was inhibited by Fab fragments of mAb 3G8. However, we found that the TSK anti-Fc gamma R mAbs do not bind to human Fc gamma RII expressed in either CHO cells or the P388D1 mouse macrophage cell line. Since the enzyme release could be inhibited by Fab fragments of mAb IV.3, we suggest that the signal transduction may require Fc gamma RII activation subsequent to crosslinking of the glycan phosphatidyl inositol-anchored Fc gamma RIII-1. These data demonstrate for the first time that polyspecific autoantibodies with Fc gamma R specificity can trigger neutrophil enzyme release via human Fc gamma RIII-1 in vitro and indicate a possible role for such autoantibodies in autoimmune inflammatory processes.

Correlation Between Cellular Functions and Morphological Changes of Human Trophoblast in Vitro

1975 ◽  
Vol 33 ◽  
pp. 600-601
Author(s):  
John C. Garancis ◽  
Robert O. Hussa ◽  
Michael T. Story ◽  
Donald Yorde ◽  
Roland A. Pattillo
Keyword(s):  
Electron Microscopy ◽  
Cellular Proliferation ◽  
Morphological Changes ◽  
Culture Fluid ◽  
Cellular Functions ◽  
Human Trophoblast ◽  
Transmission Electron ◽  
Marked Activity ◽  
Malignant Trophoblast

Human malignant trophoblast cells in continuous culture were incubated for 3 days in medium containing 1 mM N6-O2'-dibutyryl cyclic adenosine 3':5'-monophosphate (dibutyryl cyclic AMP) and 1 mM theophylline. The culture fluid was replenished daily. Stimulated cultures secreted many times more chorionic gonadotropin and estrogens than did control cultures in the absence of increased cellular proliferation. Scanning electron microscopy revealed remarkable surface changes of stimulated cells. Control cells (not stimulated) were smooth or provided with varying numbers of microvilli (Fig. 1). The latter, usually, were short and thin. The surface features of stimulated cells were considerably different. There was marked increase of microvilli which appeared elongated and thick. Many cells were covered with confluent polypoid projections (Fig. 2). Transmission electron microscopy demonstrated marked activity of cytoplasmic organelles. Mitochondria were increased in number and size; some giant forms with numerous cristae were observed.

scholarly journals Monoclonal antibody AHN-1 inhibits phagocytosis by human neutrophils

Blood ◽  
1985 ◽  
Vol 65 (2) ◽  
pp. 333-339
Author(s):  
KM Skubitz ◽  
DJ Weisdorf ◽  
PK Peterson
Keyword(s):  
Monoclonal Antibody ◽  
Lysosomal Enzyme ◽  
Surface Proteins ◽  
Superoxide Production ◽  
Human Neutrophils ◽  
Enzyme Release ◽  
Specific Monoclonal Antibody ◽  
Normal Human ◽  
Major Surface

The granulocyte-specific monoclonal antibody, AHN-1, immunoprecipitates two major surface-iodinated proteins of 105,000 and 145,000 to 150,000 daltons from normal human neutrophils. In this study, the effect of AHN- 1 on a number of neutrophil functions was evaluated in vitro. Both complement- and antibody-mediated phagocytosis were inhibited when human neutrophils were pretreated with AHN-1 and opsonized bacteria were used as targets. The inhibition of phagocytosis was specific, in that lysosomal enzyme release and chemotaxis were not altered by treatment with AHN-1. AHN-1 did inhibit superoxide production by neutrophils in response to particulate stimuli, but not in response to the soluble stimulus, 12-O-tetradecanoylphorbol-13-acetate. The data indicate that one or both of these surface proteins may be important in the process of phagocytosis. AHN-1 should be useful in isolating and further characterizing the nature of these molecules.

Paracrystalline bundles of large tubules, induced in vitro by Mebendazole in Cysticercus cellulosae

Parasitology ◽  
1981 ◽  
Vol 83 (3) ◽  
pp. 513-518 ◽  
Author(s):  
J. P. Laclette ◽  
Marie Therese Merchant ◽  
Kaethe Willms ◽  
L. Cañedo
Keyword(s):  
Electron Microscopy ◽  
Culture Medium ◽  
Morphological Changes ◽  
Bladder Wall ◽  
Secretory Cells ◽  
External Diameter ◽  
Nematode Larvae ◽  
Transmission Electron ◽  
Cysticercus Cellulosae

SUMMARYThe effect of the anthelmintic Mebendazole on Cysticercus cellulosae maintained in culture medium was studied by transmission electron microscopy. In addition to the well-known morphological changes induced by Mebendazole in other cestode and nematode larvae, it also induced the cytoplasmic appearance of paracrystalline bundles in the secretory cells of the bladder wall. These bundles were formed by groups of large parallel tubules arranged in a hexagonal-like pattern. The tubules, which had an external diameter of about 50 nm and a length that might exceed 5 μm, were surrounded by a matrix and a distance between neighbouring tubules of 80–120 nm centre to centre was estimated. The tubules were stable to colchicine and low temperature. The temporary appearance of bundles is described and some alternative explanations on their origin are advanced.

scholarly journals Differentiation of a teratocarcinoma line: preferential development of cholinergic neurons.

1981 ◽  
Vol 88 (1) ◽  
pp. 57-66 ◽  
Author(s):  
S E Pfeiffer ◽  
H Jakob ◽  
K Mikoshiba ◽  
P Dubois ◽  
J L Guenet ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Cholinergic Neurons ◽  
Carcinoma Cells ◽  
Sodium Influx ◽  
Average Cell ◽  
Transmission Electron ◽  
Neuronal Processes ◽  
Embryonic Antigen ◽  
Average Cell Volume

A line of embryonal carcinoma cells, PCC7-S, established in vitro from a spontaneous testicular teratocarcinoma, has been studied. Upon removing the cells from a low density monolayer culture system and permitting the cells to form aggregates in suspension, we observed a change of several physical and biochemical parameters: (a) reduction in average cell volume, (b) blockage and accumulation of cells in G1, (c) rise in secreted protease activity, (d) rise in acetylcholinesterase and choline acetyltransferase activities, and (e) disappearance of embryonic antigen F9. Although PCC7 aggregates did not undergo substantial morphological changes while suspended, when aggregates 4 or more days old were allowed to attach to plastic tissue culture dishes, substantial neurite outgrowth occurred over the next 1-3 d. This process was markedly enhanced by the addition to the growth medium of carboxymethylcellulose and inhibitors of DNA synthesis. Transmission electron microscopy disclosed a neurite ultrastructure consistent with that of neuronal processes. A veratridine-stimulated, tetrodotoxin-blocked sodium influx of 100 nmol/min per mg protein was also observed in these differentiated surface cultures. This cell line is discussed in terms of its utility for the study of early events leading to a commitment to cellular differentiation, as well as for the investigation of terminal differentiation to cholinergic neurons.

Ultrastructural Characterization of Platelet-Activating Factor-Stimulated Human Eosinophils from Patients with Asthma

1993 ◽  
Vol 84 (4) ◽  
pp. 391-399 ◽  
Author(s):  
Claus Kroegel ◽  
Ann Dewar ◽  
Tatsuo Yukawa ◽  
Per Venge ◽  
Peter J. Barnes ◽  
...  
Keyword(s):  
Morphometric Analysis ◽  
Morphological Changes ◽  
Shape Change ◽  
Platelet Activating Factor ◽  
Eosinophil Cationic Protein ◽  
Cell Periphery ◽  
Cationic Protein ◽  
Asthmatic Patients ◽  
Transmission Electron

1. Purified human eosinophils from asthmatic patients were stimulated with platelet-activating factor in vitro and examined for morphological changes by transmission electron and light microscopy. Changes were also evaluated by morphometric analysis and were related to the platelet-activating factor-stimulated release of granular eosinophil cationic protein. 2. Stimulation of eosinophils with platelet-activating factor induced a dose-dependent shape change, including the elongation of cells, loss of microvilli and the formation of lamellipodia. This effect was maximal at 25 min and was reversible. 3. Stimulation with platelet-activating factor also induced granule movement to the cell periphery and fusion of adjacent granules. Granules became swollen and vesiculated, whereas both the matrix and core showed evidence of solubilization. 4. There was a time-dependent secretion of eosinophilic cationic protein from human eosinophils upon stimulation with platelet-activating factor which occurred without significant lactate dehydrogenase release. 5. Morphometric analysis of the transmission electron micrographs indicated a significant reduction in cytoplasmic area after 10 min of incubation with platelet-activating factor from 39.0 ± 1.7 μm2 for untreated eosinophils to 33.2 ± 2.3 μm2 (P < 0.02) for platelet-activating factor-treated cells, underscoring the observation that the cells change from spherical to ellipsoidal. No significant increase in the perimeter of the cells was found. 6. The number of granule-profiles in platelet-activating factor-stimulated eosinophils was slightly reduced when compared with control, and an increase in granule area was observed 10 min after platelet-activating factor challenge (0.215 ± 0.011 μm2 versus 0.246 ± 0.016 μm2). 7. Human eosinophils from patients with asthma stimulated with platelet-activating factor undergo both cellular and granular alterations and reorganization which parallel the release of granular eosinophil basic protein.

Production of Peptides Inducing Chemotaxis and Lysosomal Enzyme Release in Human Neutrophils by Intestinal Bacteria in Vitro and in Vivo

1988 ◽  
Vol 23 (1) ◽  
pp. 121-128 ◽  
Author(s):  
V. S. Chadwick ◽  
D. M. Mellor ◽  
D. B. Myers ◽  
A. C. Selden ◽  
A. Keshavarzian ◽  
...  
Keyword(s):  
Lysosomal Enzyme ◽  
Intestinal Bacteria ◽  
Human Neutrophils ◽  
Enzyme Release

Iohexol-Induced Neutrophil Myeloperoxidase Release and Activation upon Contact with Vascular Stent-Graft Material: A Mechanism Contributing to the Postimplantation Syndrome?

2003 ◽  
Vol 10 (5) ◽  
pp. 958-967 ◽  
Author(s):  
Vibeke Videm ◽  
Asbjørn Ødegård ◽  
Hans Olav Myhre
Keyword(s):  
Stent Graft ◽  
Neutrophil Activation ◽  
Human Neutrophils ◽  
Maximal Response ◽  
Graft Material ◽  
Vascular Stent ◽  
Neutrophil Degranulation ◽  
Considerable Individual Variation ◽  
Vitro Effect

Purpose: To investigate whether the contrast medium iohexol alone or in combination with vascular stent-graft material induces neutrophil degranulation. Methods: Human whole blood or isolated human neutrophils were incubated with or without iohexol and vascular stent-graft material. Samples were also drawn from 5 patients undergoing diagnostic angiography using iohexol. Myeloperoxidase and lactoferrin concentrations were determined by enzyme immunoassays. Results: Both in vitro and in the patients, iohexol induced neutrophil degranulation with considerable individual variation in dose sensitivity and timing. The in vitro effect was independent of the type of anticoagulant used (ethylenediamine tetra-acetic acid, heparin, lepirudin). Experiments using isolated neutrophils showed that degranulation was independent of complement activation or platelet-derived mediators. The dose for maximal response varied from 5 to 50 mg I/mL (10.7–107.6 mg/mL iohexol). In vitro, vascular stent-graft material alone did not induce neutrophil degranulation. As compared to iohexol alone, incubation with iohexol and vascular stent-graft material in combination substantially increased the release of myeloperoxidase. Conclusions: Iohexol induces neutrophil degranulation, which is greatly enhanced when combined with vascular stent-graft material. Thus, iohexol-induced neutrophil activation may contribute to an inflammatory response following stent-graft implantation. We speculate that neutrophil activation during other procedures combining catheters and iohexol (e.g., angiography) may induce inflammation, which might have detrimental effects.

scholarly journals Antileishmanial Activity of a Linalool-Rich Essential Oil from Croton cajucara

2003 ◽  
Vol 47 (6) ◽  
pp. 1895-1901 ◽  
Author(s):  
Maria do Socorro S. Rosa ◽  
Ricardo R. Mendonça-Filho ◽  
Humberto R. Bizzo ◽  
Igor de Almeida Rodrigues ◽  
Rosangela Maria A. Soares ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Essential Oil ◽  
Mammalian Cells ◽  
Morphological Changes ◽  
Peritoneal Macrophages ◽  
Transmission Electron ◽  
Low Concentrations ◽  
Mouse Peritoneal Macrophages ◽  
Croton Cajucara

ABSTRACT The in vitro leishmanicidal effects of a linalool-rich essential oil from the leaves of Croton cajucara against Leishmania amazonensis were investigated. Morphological changes in L. amazonensis promastigotes treated with 15 ng of essential oil per ml were observed by transmission electron microscopy; leishmanial nuclear and kinetoplast chromatin destruction, followed by cell lysis, was observed within 1 h. Pretreatment of mouse peritoneal macrophages with 15 ng of essential oil per ml reduced by 50% the interaction between these macrophages and L. amazonensis, with a concomitant increase by 220% in the level of nitric oxide production by the infected macrophages. Treatment of preinfected macrophages with 15 ng of essential oil per ml reduced by 50% the interaction between these cells and the parasites, which led to a 60% increase in the amount of nitric oxide produced by the preinfected macrophages. These results provide new perspectives on the development of drugs with activities against Leishmania, as linalool-rich essential oil is a strikingly potent leishmanicidal plant extract (50% lethal doses, 8.3 ng/ml for promastigotes and 8.7 ng/ml for amastigotes) which inhibited the growth of L. amazonensis promastigotes at very low concentrations (MIC, 85.0 pg/ml) and which presented no cytotoxic effects against mammalian cells.

Leukocyte antibacterial functions are not impaired by perfluorocarbon exposure in vitro

2008 ◽  
Vol 295 (1) ◽  
pp. L134-L142 ◽  
Author(s):  
Dirk Haufe ◽  
Eva Koenigshausen ◽  
Lilla Knels ◽  
Martina Wendel ◽  
Sebastian N. Stehr ◽  
...  
Keyword(s):  
Host Defense ◽  
In Vitro Model ◽  
Polymorphonuclear Neutrophils ◽  
Human Neutrophils ◽  
Intracellular Uptake ◽  
E Coli ◽  
Transmission Electron ◽  
Membrane Bound ◽  
Vitro Model

Application of liquid, aerosolized, and vaporized perfluorocarbons (PFC) in acute lung injury has shown anti-inflammatory effects. Although this may be beneficial in states of pulmonary hyperinflammation, it also could increase susceptibility to nosocomial lung infection. We hypothesized that PFC impair cellular host defense and therefore investigated in an in vitro model the influence of perfluorohexane (PFH) on crucial mechanisms of bacterial elimination in human neutrophils and monocytes. Using scanning and transmission electron microscopy, we could show membrane-bound and ingested PFH particles that morphologically did not alter adherence and phagocytosis of Escherichia coli or leukocyte viability. The amount of adherent and phagocytosed bacteria as determined by flow cytometry was not influenced in cells only pretreated with PFH for 1 and 4 h. When PFH was present during E. coli challenge, bacterial adherence was decreased in polymorphonuclear neutrophils, but respective intracellular uptake was not impaired and was even significantly promoted in monocytes. Overall, E. coli-induced respiratory burst capacity was not reduced by PFH. Our findings provide evidence that key functions of innate host defense are not compromised by PFH treatment in vitro.

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