scholarly journals THE CYTOTOXIC EFFECT OF MOUSE MACROPHAGES ON SYNGENEIC AND ALLOGENEIC ERYTHROCYTES

1973 ◽  
Vol 137 (3) ◽  
pp. 807-820 ◽  
Author(s):  
H. Melsom ◽  
R. Seljelid
Keyword(s):  
Cytotoxic Effect ◽  
Peritoneal Macrophages ◽  
Target Cells ◽  
Activated Macrophages ◽  
Con A ◽  
Osmotic Effect ◽  
Prolonged Cultivation ◽  
Cytotoxic Reaction ◽  
Mouse Macrophages ◽  
Mouse Peritoneal Macrophages

A cytotoxic effect of mouse peritoneal macrophages against syngeneic and allogeneic erythrocytes was demonstrated by isotope release and release of hemoglobin. The cytotoxic effect was dependent on the contact between viable, activated macrophages and target cells. Activation was accomplished by prolonged cultivation of macrophages and by the presence of Zn++ and Con-A. Immunization did not prove necessary. Morphological observations as well as experiments with various salt concentrations indicate that the cytotoxic reaction may involve some kind of osmotic effect upon the target cells.

scholarly journals Candidacidal activity of mouse macrophages in vitro

1980 ◽  
Vol 29 (2) ◽  
pp. 477-482 ◽  
Author(s):  
P K Maiti ◽  
R Kumar ◽  
L N Mohapatra
Keyword(s):  
Peritoneal Macrophages ◽  
Mouse Serum ◽  
Activated Macrophages ◽  
Immune Mouse ◽  
Mouse Macrophages ◽  
In Vitro Exposure ◽  
Candida Infections ◽  
Mouse Peritoneal Macrophages ◽  
Immune Mouse Serum

Mouse peritoneal macrophages were infected in vitro with Candida albicans, and the phagocytic and candidacidal activities were estimated by microscopic examination of Giemsa-stained cells. Activated macrophages obtained from either BCG-vaccinated animals or by in vitro exposure of normal macrophages to phytohemagglutinin-induced lymphokines exhibited higher phagocytic and candidacidal activities than did normal macrophages. However, activated macrophages obtained by in vitro exposure of macrophages to candida-induced lymphokines exhibited the highest phagocytic and candidacidal activities. The incorporation of immune mouse serum into the culture medium also enhanced the phagocytic and candidacidal activities of the normal macrophages but failed to improve the function of the activated macrophages. These results suggest that both activated macrophages and antibodies may be required for controlling candida infections in mice.

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.

Structural heterogeneity of endocytic membranes in macrophages as revealed by the cholesterol probe, filipin

1981 ◽  
Vol 51 (1) ◽  
pp. 95-107
Author(s):  
R. Montesano ◽  
P. Vassalli ◽  
L. Orci
Keyword(s):  
Structural Heterogeneity ◽  
Cholesterol Content ◽  
Peritoneal Macrophages ◽  
Con A ◽  
Coated Pits ◽  
Mechanisms Of Formation ◽  
Smooth Membrane ◽  
Endocytic Vesicles ◽  
Cytochemical Technique ◽  
Mouse Peritoneal Macrophages

The polyene antibiotic, filipin, by specifically interacting with cholesterol, produces approximately 25-nm protuberances (filipin-sterol complexes) in freeze-fractured membranes, and the addition of filipin to aldehyde fixatives has been recently introduced as a cytochemical technique for the localization of cholesterol in cell membranes. In a previous study we showed that, in fibroblasts filipin-sterol complexes are absent from endocytic coated pits. To establish whether the absence of filipin-sterol complexes is a phenomenon restricted to coated pits or is correlated with endocytosis in general, we applied the filipin probe to cultured mouse peritoneal macrophages, in which different forms of endocytosis take place. The macrophages were incubated with bovine albumin or concanavalin A (Con A) to induce pinocytosis, and with heat-killed straphylococci or opsonized erythrocytes to induce phagocytosis, then fixed in glutaraldehyde/filipin and freeze-fractured. Filipin-sterol complexes were plentiful on the plasma membrane, on the smooth-membrane invaginations and vesicles induced by albumin, on the large endocytic vacuoles induced by Con A, and on the membrane of phagosomes but, in contrast, they were absent from coated pits and vesicles, as well as from coated segments of invagination or vesicles. These results indicate that the membranes involved in different types of endocytosis do not react in the same way with filipin and may, therefore, have a different cholesterol content. This could reflect different mechanisms of formation for the various types of endocytic vesicles.

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.

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 CONCANAVALIN A ON MOUSE PERITONEAL MACROPHAGES

1974 ◽  
Vol 140 (5) ◽  
pp. 1364-1386 ◽  
Author(s):  
Paul J. Edelson ◽  
Zanvil A. Cohn
Keyword(s):  
Concanavalin A ◽  
Peritoneal Macrophages ◽  
Surface Marker ◽  
Marker Enzyme ◽  
Apparent Effect ◽  
Con A ◽  
Cytoplasmic Vesicles ◽  
Secondary Lysosomes ◽  
Mouse Peritoneal Macrophages ◽  
Pinocytic Vesicles

Concanavalin A (Con A) binds to saccharide residues on the mouse peritoneal macrophage plasma membrane and stimulates extensive pinocytic interiorization of the membrane. The overall pinocytic rate is increased 3.5–4.5 times by the addition of Con A, and the surface marker enzyme adenosine triphosphatase can be identified histochemically in association with the cytoplasmic vesicles generated after exposure of the cells to Con A. Once formed, these pinocytic vesicles may persist for several days and fail to show morphologic evidence of fusion with primary or preformed secondary lysosomes. There is no apparent effect on the capacity of the macrophage to ingest either latex particles or IgG-coated SRBC administered either simultaneously with or subsequent to the Con A.

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.

Infiltration and survival: the behaviour of normal, invasive cells implanted to the developing chick wing

1979 ◽  
Vol 40 (1) ◽  
pp. 257-270
Author(s):  
C. Tickle ◽  
A. Crawley
Keyword(s):  
Endothelial Cells ◽  
Plasminogen Activator ◽  
Blood Cells ◽  
Peritoneal Macrophages ◽  
Neutrophil Granulocytes ◽  
Activated Macrophages ◽  
Wing Bud ◽  
Mesenchyme Cells ◽  
Mouse Peritoneal Macrophages ◽  
Mouse Lymphocytes

The invasiveness of mouse lymphocytes and thymocytes, rabbit peritoneal neutrophil granulocytes (PMNs), mouse peritoneal macrophages (both activated and non-activated) and pig endothelial cells was assayed by implanting these cells to the chick wing bud. Cells of each type moved into the wing mesenchyme, although activated macrophages invaded poorly. PMNs were the most invasive cells and had moved well into the limb after only a few hours. PMNs, lymphocytes and thymocytes were ingested by wing mesenchyme cells. Endothelial cells, however, ingested chick blood cells. The implanted cells showed differences in ability to survive in the limb: PMNs disappeared rapidly, lymphocytes and thymocytes sometimes persisted for 24 h, while grafts of macrophages and endothelial cells were present at 24 h. Mechanisms which might be involved in the invasiveness of these cells, and also in their different abilities to survive in the chick wing, are discussed with particular reference to the production of plasminogen activator.

scholarly journals THE INTERACTION OF SOLUBLE HORSERADISH PEROXIDASE WITH MOUSE PERITONEAL MACROPHAGES IN VITRO

1972 ◽  
Vol 55 (1) ◽  
pp. 186-204 ◽  
Author(s):  
Ralph M. Steinman ◽  
Zanvil A. Cohn
Keyword(s):  
Horseradish Peroxidase ◽  
Culture Medium ◽  
Peritoneal Macrophages ◽  
Wide Range ◽  
Mouse Macrophages ◽  
Secondary Lysosomes ◽  
Endocytic Vesicles ◽  
In Vitro Interaction ◽  
Mouse Peritoneal Macrophages

The in vitro interaction of soluble horseradish peroxidase (HRP) with homogeneous mono layers of mouse macrophages has been studied using sensitive biochemical and cytochemical techniques. The compartmentalization of HRP in extracellular and intracellular sites has been quantitatively evaluated. A significant fraction is bound to a serum-derived layer, which coats the surface of culture vessels and may be removed by appropriate washes. Macrophages interiorize HRP as a solute in pinocytic vesicles without appreciable binding of the glycoprotein to the plasma membrane. Uptake is directly proportional to the concentration of HRP in the culture medium. 1 x 106 cells ingest 0.0025% of the administered load per hr over a wide range of concentrations. Cytochemically, all demonstrable HRP is sequestered within the endocytic vesicles and secondary lysosomes of the vacuolar apparatus. After uptake, the enzymatic activity of HRP is inactivated exponentially with a half-life of 7–9 hr, until enzyme is no longer detectable. When macrophages have pinocytosed trace-labeled HRP-125I, cell-associated isotope disappears with a t ½ of 20–30 hr and they release monoiodotyrosine-125I into the culture medium. We were unable to obtain evidence that significant amounts of HRP (>2%) can be exocytosed after uptake, can exist intact on the cell surface, or can be digested extracellularly. It is difficult to reconcile these observations with several of the postulated mechanisms whereby macrophages are thought to play a prominent role in the induction of an immune response.

scholarly journals THE INFLUENCE OF ERYTHROPHAGOCYTOSIS ON THE INTERACTION OF MACROPHAGES AND SALMONELLA IN VITRO

1966 ◽  
Vol 124 (2) ◽  
pp. 173-183 ◽  
Author(s):  
Fred A. Gill ◽  
Donald Kaye ◽  
Edward W. Hook
Keyword(s):  
Salmonella Typhimurium ◽  
Peritoneal Macrophages ◽  
Cytotoxic Action ◽  
No Inhibition ◽  
Mouse Macrophages ◽  
Mouse Erythrocyte ◽  
Mouse Peritoneal Macrophages

Phagocytosis and killing of Salmonella typhimurium by mouse peritoneal macrophages was inhibited when the bacteria and antibody-coated homologous erythrocytes or heterologous erythrocytes were simultaneously exposed to macrophages in vitro. No inhibition of phagocytosis or killing was observed in experiments employing uncoated or disrupted antibody-coated homologous erythrocytes. Degradation of S. typhimurium as measured by the loss of fluorescence from intracellular salmonella coated with fluorescein-labeled antibody was inhibited in macrophages which had previously ingested antibody-coated homologous erythrocytes. Anti-mouse-erythrocyte serum was found to have a cytotoxic action on mouse macrophages. However, the viability of macrophages was not altered by phagocytosis of antibody-coated homologous erythrocytes or uncoated heterologous erythrocytes.

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