scholarly journals Primary amines induce selective release of lysosomal enzymes from mouse macrophages

1980 ◽  
Vol 188 (3) ◽  
pp. 933-936 ◽  
Author(s):  
D W Riches ◽  
D R Stanworth
Keyword(s):  
Chain Length ◽  
Lysosomal Enzyme ◽  
Lysosomal Enzymes ◽  
Peritoneal Macrophages ◽  
Primary Amines ◽  
Aliphatic Chain ◽  
Enzyme Release ◽  
Mouse Macrophages ◽  
Aliphatic Diamines ◽  
Mouse Peritoneal Macrophages

Cultured mouse peritoneal macrophages were found to release substantial amounts of lysosomal beta-glucuronidase and beta-glactosidase activites when exposed to millimolar concentrations of various primary aliphatic monoamines. With methylamine, ethylamine, propylamine and butylamine, lysosomal enzyme release was selective, but further increases in the aliphatic chain length resulted in the compounds becoming lytic. By contrast, structurally related primary aliphatic diamines proved to be inactive at inducing both selective and lytic lysosomal-enzyme discharge.

scholarly journals The selective release of phospholipase A2 by resident mouse peritoneal macrophages

1981 ◽  
Vol 200 (2) ◽  
pp. 441-444 ◽  
Author(s):  
P D Wightman ◽  
M E Dahlgren ◽  
P Davies ◽  
R J Bonney
Keyword(s):  
Phospholipase A2 ◽  
Phospholipase C ◽  
Lysosomal Enzyme ◽  
Enzyme Activities ◽  
Lysosomal Enzymes ◽  
Culture Medium ◽  
Peritoneal Macrophages ◽  
Cellular Activity ◽  
Mouse Peritoneal Macrophages ◽  
Phospholipase A2 Activity

Resident mouse peritoneal macrophages have three phospholipase activities: a phospholipase A2 active at pH 4.5, a Ca2+-dependent phospholipase A2 active at pH 8.5 and a phosphatidylinositol-specific phospholipase C activity. When macrophages are exposed to zymosan in culture, the cellular activity of pH-4.5 phospholipase A2 is diminished in a manner dependent on zymosan concentration and time of exposure, whereas the cellular activities of pH-8.5 phospholipase A2 and phospholipase C remain unchanged. The depletion of pH-4.5 phospholipase A2 activity from the cell is paralleled by a quantitative recovery of this activity in the culture medium in a manner similar to the cellular depletion and extracellular recovery of two lysosomal enzymes. This release is specifically elicited by an inflammatory substance such as zymosan, since macrophages incubated with 6 micrometer latex spheres retain pH-4.5 phospholipase A2 activity and lysosomal enzyme activities intracellularly.

Release of β-glucuronidase from peritoneal macrophages of normal and endotoxin-tolerant mice

1979 ◽  
Vol 25 (11) ◽  
pp. 1245-1251 ◽  
Author(s):  
Stephen L. Snyder ◽  
Sharyn K. Eklund ◽  
Richard I. Walker
Keyword(s):  
Lysosomal Enzyme ◽  
Lysosomal Enzymes ◽  
Lethal Dose ◽  
In Vitro Release ◽  
Peritoneal Macrophages ◽  
Endotoxin Shock ◽  
Enzyme Release ◽  
Endotoxin Challenge

Lysosomal enzyme release from cells involved in inflammatory response could play a central role in the pathogenesis of endotoxin shock. Therefore we have studied the release of the lysosomal enzyme, β-glucuronidase, from peritoneal macrophages obtained from normal and endotoxin-tolerant B6CBF1 mice both before and after challenge with lethal doses of endotoxin. Unstimulated cells from tolerant mice spontaneously released a smaller percentage of their total β-glucuronidase content in culture than cells from normal mice during a 5-h incubation period. In support of the lysosomal enzyme release hypothesis, it was found that the in vitro release of β-glucuronidase was accelerated when cells were collected from the mouse peritoneum 3 h after i.v. challenge with a lethal dose (1.0 mg) of endotoxin. The increased in vitro "leakiness" of peritoneal macrophages following endotoxin challenge was less marked when tolerance was induced in mice by prior repeated injections of endotoxin. Furthermore, measurements of the total enzyme activities of peritoneal cells revealed a significant reduction in the β-glucuronidase content of cells from normal mice 3 h after endotoxin challenge but no such decrease for cells from tolerant mice. These results suggest that macrophages in endotoxin-sensitive mice release their lysosomal enzymes in vivo during endotoxemia, whereas cells found in tolerant mice do not.In related experiments, the phagocytosis of latex particles and inhibition of bacterial growth by macrophages from normal and tolerant mice were compared. These studies suggest that cells from tolerant mice may also release a smaller percentage of their lysosomal enzymes during phagocytosis.

scholarly journals Phagolysosome formation in normal and colchicine-treated macrophages.

1975 ◽  
Vol 142 (4) ◽  
pp. 903-913 ◽  
Author(s):  
E L Pesanti ◽  
S G Axline
Keyword(s):  
Acid Phosphatase ◽  
Lysosomal Enzyme ◽  
Peritoneal Macrophages ◽  
Cell Type ◽  
Inert Particles ◽  
Time Periods ◽  
Mouse Peritoneal Macrophages ◽  
Over Time

Intracellular lysosomal fusion has been evaluated in cultivated mouse peritoneal macrophages by measurement of transfer of acid phosphatase to polyvinyltoluene (PVT)-containing phagolysosomes. Enzyme transfer was found to be directly and significantly related to the uptake of PVT and to be independent of time allowed for phagolysosome formation over time periods of 15 min to 18 h. In addition, the extent of transfer of lysosomal enzyme to phagolysosomes was unaffected by treatment of the cells with 10(-6) M colchicine, a dose which eradicates morphologically identifiable microtubules in this cell type within 2 h. The data indicate that intracellular fusion of lysosomes with phagosomes in the macrophage does not require formed microtubules and suggest that fusion occurs promptly after interiorization of inert particles.

Transport of phagosomes in mouse peritoneal macrophages

1989 ◽  
Vol 94 (1) ◽  
pp. 143-153
Author(s):  
A. Toyohara ◽  
K. Inaba
Keyword(s):  
Cytochalasin B ◽  
Large Diameter ◽  
Sulfate Solution ◽  
Peritoneal Macrophages ◽  
Transport Systems ◽  
Perinuclear Region ◽  
Microtubule Network ◽  
Mouse Macrophages ◽  
Hank’S Solution ◽  
Mouse Peritoneal Macrophages

Mouse macrophages were elicited by the peritoneal injection of chondroitin sulfate solution, harvested and purified, and used as experimental materials. Small and large (diameter: 0.9 microns and 3.0 microns, respectively) polystyrene beads (PB) were used as ingested particles. When the macrophages were incubated with Hank's solution containing small or large PB for 30 min, the phagosomes containing small or large PB were usually randomly distributed. When the macrophages were further incubated for 45 min in PB-free medium, both small and large phagosomes containing PB accumulated at the perinuclear region. The transport of large phagosomes containing 3.0 microns PB was inhibited by cytochalasin B, but not by vinblastine or podophyllotoxin. Conversely, the transport of small phagosomes containing 0.9 microns PB was not inhibited by cytochalasin B but was inhibited by vinblastine or podophyllotoxin. Immunofluorescence microscopy showed that the small phagosomes appeared to accumulate at the central region of the microtubule network. The large phagosomes, on the other hand, appeared to be surrounded by actin-rich cytoplasm, and in some cells actin filament-like structures could be seen around large phagosomes. These results suggest that there are two different transport systems of phagosomes in macrophages. Phagosomes smaller than 0.9 microns in diameter are, probably, mainly transported to the perinuclear region by a microtubule-based motility system and those larger than 3.0 microns in diameter by an actin-based mechanism. It was observed electron-microscopically that accumulated phagosomes containing PB could fuse with each other and form larger phagosomes.

scholarly journals IN VITRO STUDIES ON THE INTERACTION BETWEEN MOUSE PERITONEAL MACROPHAGES AND STRAINS OF SALMONELLA AND ESCHERICHIA COLI

1960 ◽  
Vol 112 (2) ◽  
pp. 403-417 ◽  
Author(s):  
Charles Jenkin ◽  
Baruj Benacerraf
Keyword(s):  
Escherichia Coli ◽  
Normal Mouse ◽  
Peritoneal Macrophages ◽  
In Vitro Studies ◽  
Experimental Conditions ◽  
Mouse Plasma ◽  
Virulent Strains ◽  
Mouse Macrophages ◽  
Mouse Peritoneal Macrophages

Virulent strains of Salmonella opsonized with normal mouse plasma are never phagocytosed as well as avirulent strains. The virulent strains of Salmonella phagocytosed after opsonization with normal mouse plasma are able to multiply within normal mouse peritoneal macrophages, whereas under similar experimental conditions the avirulent strains are killed. When virulent strains of Salmonella are opsonized with specific antiserum or plasma from BCG-infected mice, they are treated by normal mouse macrophages as if they were avirulent. Virulent bacteria opsonized with BCG plasma are phagocytosed and killed better by peritoneal macrophages from BCG-infected mice, than peritoneal macrophages from normal mice.

The Induction of Lysosomal Enzyme Release from Leucocytes of Normal and Emphysematous Subjects and the Effects of Tobacco Smoke upon Phagocytosis

1980 ◽  
Vol 58 (5) ◽  
pp. 403-409 ◽  
Author(s):  
D. C. S. Hutchison ◽  
R. Desai ◽  
D. Bellamy ◽  
H. Baum
Keyword(s):  
Cigarette Smoke ◽  
Lysosomal Enzyme ◽  
Lysosomal Enzymes ◽  
Soluble Fraction ◽  
Normal Subjects ◽  
Polymorphonuclear Leucocytes ◽  
Enzyme Release ◽  
Elastic Tissue ◽  
Latex Particles ◽  
Respiratory Inhibitor

1. The lysosomal enzymes of circulating polymorphonuclear leucocytes contain a potent elastase; release of this enzyme within the lung is thought to be responsible for the destruction of elastic tissue in pulmonary emphysema. 2. The release of lysosomal enzymes from blood leucocytes of normal and emphysematous subjects during phagocytosis of particulate material was studied In vitro. Acid phosphatase and acid ribonuclease were used as markers of lysosomal enzyme release, no sufficiently sensitive assay for elastase being available. Cigarette smoke was separated into ‘particulate’ and ‘soluble’ fractions. In a preliminary study, the particulate fraction stimulated enzyme release; in the experiments reported here, latex particles were used to produce this effect. 3. Approximately one-third of the total lysosomal enzyme content was released to the exterior of the cell during phagocytosis of latex particles. In this respect there was no difference between normal and emphysematous subjects. 4. The effects of the non-particulate soluble fraction of cigarette smoke on phagocytosis-induced enzyme release were studied. This fraction inhibited enzyme release from polymorphonuclear leucocytes of normal subjects but not from those of emphysematous patients. When the ‘cigarette-smoke solution’ was replaced by the respiratory inhibitor, antimycin A, a similar inhibition of enzyme release occurred. The inhibition of phagocytosis in cells of normal subjects is presumed to be due to a respiratory inhibitor such as carbon monoxide in the soluble fraction of the smoke. We postulate that the polymorphonuclear leucocytes of emphysematous patients are adapted to hypoxic conditions so that inhibition of enzyme release does not occur.

scholarly journals THE INTERACTION IN VITRO OF MYCOPLASMA PULMONIS WITH MOUSE PERITONEAL MACROPHAGES AND L-CELLS

1971 ◽  
Vol 133 (2) ◽  
pp. 231-259 ◽  
Author(s):  
Thomas C. Jones ◽  
James G. Hirsch
Keyword(s):  
Tritiated Thymidine ◽  
Calf Serum ◽  
Peritoneal Macrophages ◽  
Mycoplasma Pulmonis ◽  
Cultural Conditions ◽  
Low Concentrations ◽  
Mouse Macrophages ◽  
Similar Growth ◽  
Mouse Peritoneal Macrophages

Methods have been devised for establishing infection in vitro of mouse macrophages and fibroblasts with Mycoplasma pulmonis. The mycoplasmas attached to the cells and under appropriate cultural conditions grew into a lawn of microorganisms covering most of the cell surface. The mycoplasmas grew abundantly on fibroblasts cultured in minimal essential medium containing 20% fetal calf serum; supplementation of this medium with heart infusion broth was necessary to obtain similar growth on macrophages. The infection of these cells appeared to be essentially an extracellular process; only rarely were partially degraded mycoplasmas seen with phagocytic vacuoles. The addition to heavily infected macrophage cultures of low concentrations of anti-mycoplasma antibody stimulated rapid, massive phagocytosis of the surface microorganisms. In sharp contrast, the same antiserum had no discernable effect on the mycoplasma-fibroblast relationship. The antibody effect in the macrophage system was apparently a direct opsonic one rather than an indirect result of microbial killing, since the mycoplasmas in macrophage or fibroblast cultures incorporated labelled thymidine into DNA after the addition of antiserum to the medium. The phagocytic event and the subsequent fate of the mycoplasmas were studied in detail after the addition of antibody to the macrophage cultures. Phase-contrast cinemicrophotography revealed membrane ruffles surrounding the surface mycoplasmas and disappearance from view of the organisms; 10–30 min later translucent grapelike clusters were seen in large phagocytic vacuoles. On electronmicroscopic study the surface mycoplasmas were surrounded by pincers-like projections of the macrophage. Numerous mycoplasmas were seen in phagocytic vacuoles; in the early minutes after the addition of antibody the intracellular mycoplasmas appeared normal, but within 2 hr they appeared partially degraded with a central electron-lucent area and electron-opaque deposits at the microbial cell margin. 24 hr after the addition of antiserum, digestion of the mycoplasmas was nearly complete; the cells appeared normal except for large residual bodies composed of amorphous moderately dense material and increased lipid deposits. Degradation of mycoplasmas within macrophages was also studied using infected cultures in which the mycoplasmas, but not the macrophages, had incorporated tritiated thymidine into DNA. The appearance of large amounts of acid-soluble radiolabel after phagocytosis stimulated by antibody confirmed the degradation of the intracellular mycoplasmas.

scholarly journals Effects of cytochalasin B on actin and myosin association with particle binding sites in mouse macrophages: implications with regard to the mechanism of action of the cytochalasins.

1981 ◽  
Vol 91 (2) ◽  
pp. 373-384 ◽  
Author(s):  
R G Painter ◽  
J Whisenand ◽  
A T McIntosh
Keyword(s):  
Binding Sites ◽  
Cytochalasin B ◽  
Peritoneal Macrophages ◽  
Immunofluorescent Staining ◽  
Transmission Electron ◽  
Mouse Macrophages ◽  
Particle Binding ◽  
Punctate Pattern ◽  
Particle Ingestion ◽  
Mouse Peritoneal Macrophages

The intracellular distribution of F-actin and myosin has been examined in mouse peritoneal macrophages by immunofluorescence microscopy. In resting, adherent cells, F-actin was distributed in a fine networklike pattern throughout the cytoplasm. Myosin, in contrast, was distributed in a punctate pattern. After treatment with cytochalasin B (CB), both proteins showed a coarse punctate pattern consistent with a condensation of protein around specific foci. After CB-pretreated cells were exposed to opsonized zymosan particles, immunofluorescent staining for F-actin and myosin showed an increased staining under particle binding sites. Transmission electron microscope (TEM) examination of whole-cell mounts of such preparations revealed a dense zone of filaments beneath the relatively electron-translucent zymosan particles. At sites where particles had detached during processing, these filament-rich areas were more clearly delineated. At such sites dense arrays of filaments that appeared more or less randomly oriented were apparent. The filaments could be decorated with heavy meromyosin, suggesting that they were composed, in part, of F-actin and were therefore identical to the structures giving rise to the immunofluorescence patterns. After viewing CB-treated preparations by whole-mount TEM, we examined the cells by scanning electron microscopy (SEM). Direct SEM comparison of the filament-rich zones seen by TEM showed that these structures resulted from the formation of short lamellipodial protrusions below the site of particle binding. Electron micrographs of thin-sectioned material established that these lamellipodial protrusions were densely packed with microfilaments that were in part associated with the cytoplasmic surface of the plasma membrane. The formation of particle-associated lamellipodia did not appear to represent merely a slower rate of ingestion in the presence of CB, because they formed within minutes of particle contact with the cell membrane and were not followed by particle ingestion even after a 1-h or longer incubation. Furthermore, their formation required cellular energy. These results suggest that cytochalasin B blocks phagocytosis of large particles by affecting the distances over which any putative actomyosin-mediated forces are generated.

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