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.

scholarly journals MECHANISMS OF ACQUIRED RESISTANCE IN MOUSE TYPHOID

1966 ◽  
Vol 124 (4) ◽  
pp. 585-600 ◽  
Author(s):  
R. V. Blanden ◽  
G. B. Mackaness ◽  
F. M. Collins
Keyword(s):  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Normal Mouse ◽  
Acquired Resistance ◽  
Peritoneal Macrophages ◽  
Immune Serum ◽  
Humoral Factors ◽  
Intraperitoneal Route ◽  
Mouse Peritoneal Macrophages

Experiments in vitro comparing normal mouse peritoneal macrophages with cells from Salmonella typhimurium-infected mice have shown that the "immune" macrophages have conspicuously enhanced microbicidal properties. Whereas normal macrophages could inactivate only 50 to 60% of intracellular S. typhimurium pretreated with immune serum, cells from infected animals killed virtually all ingested organisms and did so at an accelerated rate. Macrophages from Listeria monocytogenes-infected mice were shown to possess similarly enhanced microbicidal activity against S. typhimurium. Furthermore, the growth of S. typhimurium in the liver and spleen was more effectively restricted in Listeria-infected mice than in animals vaccinated with heat-killed S. typhimurium, even though the Listeria-infected animals possessed no demonstrable cross-reacting antibody to S. typhimurium. The lack of resistance in the mice vaccinated with heat-killed organisms could not be attributed to any deficiency of humoral factors, since the serum from these animals was as effective at promoting phagocytosis and killing by macrophages as serum from actively infected (and demonstrably resistant) mice. Conversely, Salmonella-infected mice were totally resistant to intravenous challenge with L. monocytogenes. The level of resistance in individual animals was related to the numbers of residual Salmonellae remaining in the tissues; mice with heavier residual infections being the more resistant. Specific antiserum from mice vaccinated with heat-killed S. typhimurium was found to be significantly protective only when the intraperitoneal route of challenge was employed. The foregoing studies have been interpreted to mean that enhancement of the microbicidal ability of macrophages is the mechanism of major importance in acquired resistance to S. typhimurium infection in mice.

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 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 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.

scholarly journals THE INTERACTION OF PARTICULATE HORSERADISH PEROXIDASE (HRP)-ANTI HRP IMMUNE COMPLEXES WITH MOUSE PERITONEAL MACROPHAGES IN VITRO

1972 ◽  
Vol 55 (3) ◽  
pp. 616-634 ◽  
Author(s):  
Ralph M. Steinman ◽  
Zanvil A. Cohn
Keyword(s):  
Horseradish Peroxidase ◽  
Immune Complexes ◽  
Half Life ◽  
Peritoneal Macrophages ◽  
Surface Complexes ◽  
Mouse Macrophages ◽  
Membrane Bound ◽  
Mouse Peritoneal Macrophages

The uptake, distribution, and fate of particulate horseradish peroxidase (HRP)-anti HRP aggregates has been studied in homogeneous monolayers of mouse macrophages in vitro. Macrophages rapidly interiorize the immune complexes after binding to the cell surface. The rate of interiorization is maximal for complexes formed in a broad zone of 4-fold antibody excess to equivalence and corresponds to a rate of 10% of the administered load/106 cells per hour. This rate is 4000-fold greater than the uptake of soluble HRP. The binding and endocytosis of HRP-anti HRP by macrophages is mediated by the trypsin insensitive Fc receptor. Cytochemically, intracellular HRP is localized within membrane bound vacuoles. After uptake of HRP, the enzymatic activity is degraded exponentially with a half-life of 14–18 hr until enzyme is no longer detectable. This half-life is twice as long as that previously observed for soluble uncomplexed HRP and is related to the combination of HRP with anti-HRP rather than the absolute amounts of enzyme or antibody ingested. The half-life of HRP-125I was 30 hr. Exocytosis of cell associated enzyme or TCA precipitable counts was not detected, nor were persistent surface complexes demonstrable. The extensive capacity of macrophages to interiorize and destroy large amounts of antigen after the formation of antibody illustrates a role of this cell in the efferent limb of the immune response.

scholarly journals Uptake and degradation of mast-cell granules by mouse peritoneal macrophages

1979 ◽  
Vol 182 (1) ◽  
pp. 189-193 ◽  
Author(s):  
U Lindahl ◽  
H Pertoft ◽  
R Seljelid
Keyword(s):  
Mast Cell ◽  
Peritoneal Macrophages ◽  
Pulse Labelling ◽  
Sulphated Polysaccharides ◽  
Mouse Macrophages ◽  
Mouse Peritoneal Macrophages

35S-labelled mast-cell granules isolated from mouse mastocytomas were added to mouse macrophages in vitro. The granules were avidly phagocytosed, and subsequently the radioactivity was released to the medium as inorganic [35S]sulphate. After pulse-labelling, a total of about 80% of the cell-associated radioactivity was thus released in the course of 24 h, indicating an extensive breakdown of the sulphated polysaccharides, mainly heparin, present in the granules. The uptake of the mast-cell granules caused pronounced, but reversible, spreading of the macrophages.

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.

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