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 THE UPTAKE AND DIGESTION OF IODINATED HUMAN SERUM ALBUMIN BY MACROPHAGES IN VITRO

1967 ◽  
Vol 126 (5) ◽  
pp. 941-958 ◽  
Author(s):  
Barbara A. Ehrenreich ◽  
Zanvil A. Cohn
Keyword(s):  
Human Serum Albumin ◽  
Peritoneal Macrophages ◽  
Radioactive Material ◽  
In Vitro Cultivation ◽  
Cultivation Conditions ◽  
Secondary Lysosomes ◽  
Mouse Peritoneal Macrophages ◽  
Reduced Temperature ◽  
Pinocytic Vesicles

Mouse peritoneal macrophages take up I*-HSA from their medium during in vitro cultivation. Conditions which promote I*-HSA uptake are the same as those which stimulate formation of pinocytic vesicles. Autoradiography of cells pulsed with 125I-HSA showed that intracellular isotope is localized in perinuclear granules, or secondary lysosomes. Following a pulse of 125I-HSA, intracellular radioactivity decreases and the amount of TCA-soluble isotope in the medium increases correspondingly. About 50% of the intracellular isotope is lost in 5 hr. The release of isotope from pulsed cells is not inhibited by parafluorophenylalanine, 2,4-dinitrophenol or by a reduction of the serum concentration of the medium. However, the processing of ingested 125I-HSA is reversibly inhibited by reduced temperature. The TCA-soluble radioactive material excreted by pulsed macrophages was identified as monoiodotyrosine.

Participation of concanavalin A binding sites in the interaction between Trypanosoma cruzi and macrophages

1983 ◽  
Vol 62 (1) ◽  
pp. 287-299
Author(s):  
M.N. Meirelles ◽  
A. Martinez-Palomo ◽  
T. Souto-Padron ◽  
W. De Souza
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Trypanosoma Cruzi ◽  
Uniform Distribution ◽  
Concanavalin A ◽  
Binding Sites ◽  
Peritoneal Macrophages ◽  
Untreated Mouse ◽  
Mouse Peritoneal Macrophages

Untreated mouse peritoneal macrophages as well as macrophages treated with concanavalin A (ConA) were incubated in the presence of untreated or ConA-treated epimastigotes and trypomastigotes of Trypanosoma cruzi. Treatment of epimastigotes or trypomastigotes with ConA increased or decreased their uptake by macrophages, respectively. Treatment of their macrophages with ConA reduced by 70% and increased by five times the ingestion of epimastigotes and trypomastigotes, respectively. These results are discussed in relation to previous studies on the mobility of ConA receptors in the membrane of the parasite. Using fluorescein- or ferritin-labelled ConA we observed that ConA binding sites located on the plasma membrane of macrophages are internalized during endocytosis of T. cruzi, and observed in association with the membrane of the endocytic vacuole. Vacuoles without parasites showed a uniform distribution of ConA binding sites, while these sites were distributed in patches in vacuoles containing parasites. These results, in association with others previously reported, suggest the involvement of glycoproteins and/or glycolipids localized on the cell surface of T. cruzi and macrophages during the T. cruzi-macrophage interaction.

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 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 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 5'-Nucleotidase activity of mouse peritoneal macrophages. II. Cellular distribution and effects of endocytosis.

1976 ◽  
Vol 144 (6) ◽  
pp. 1596-1608 ◽  
Author(s):  
P J Edelson ◽  
Z A Cohn
Keyword(s):  
Plasma Membrane ◽  
Concanavalin A ◽  
Diazonium Salt ◽  
Peritoneal Macrophages ◽  
Sulfanilic Acid ◽  
Nucleotidase Activity ◽  
Intracellular Pool ◽  
Enzyme Degradation ◽  
Cytoplasmic Vesicles ◽  
Serum Free Conditions

The diazonium salt of sulfanilic acid (DASA) can inactivate about 80% of the total 5'-nucleotidase of viable macrophages. The remaining 20% can be inactivated if the cells are first lysed in detergent, and presumably represents an intracellular pool of 5'-nucleotidase. The bulk of this pool may represent cytoplasmic vesicles derived from plasma membrane by endocytosis. This internal compartment is expanded up to threefold immediately after the cells have ingested a large latex load. This is consistent with previous observations on the internalization of 5'-nucleotidase in latex phagosomes. In latex-filled cells this intracellular pool of enzyme is inactivated over a few hours, and the cells then slowly increase their enzyme activity to nearly normal levels. However, 24 h after latex ingestion the metabolism of 5'-nucleotidase in these recovered cells is abnormal, as the rate of enzyme degradation is about twice the normal rate, and the DASA-insensitive enzyme pool in these cells is strikingly diminished. This may reflect effects of the accumulated indigestible particles on the fate of incoming pinocytic vesicles or on newly synthesized plasma membrane precursor. Another endocytic stimulus, concanavalin A, also reduces the total cell 5'-nucleotidase activity. This effect, which is time and temperature dependent, can be prevented by the competitive sugar alpha-methyl mannose. The concanavalin A inhibition can be reversed in the absence of new protein synthesis or in cells cultivated in serum-free conditions. It is not known whether the effect of concanavalin A on 5'-nucleotidase depends upon the interiorizaiton of plasma membrane or is strictly associated with events at the cell surface.

Quantitative analysis of surface marker densities after exposure of T-cells to concanavalin A (Con A): A sensitive early index of cellular activation

1991 ◽  
Vol 133 (2) ◽  
pp. 519-525 ◽  
Author(s):  
Moshe Shabtai ◽  
Kazimierz Malinowski ◽  
Wayne C. Waltzer ◽  
Christopher Pullis ◽  
Audrey P. Raisbeck ◽  
...  
Keyword(s):  
T Cells ◽  
Quantitative Analysis ◽  
Concanavalin A ◽  
Surface Marker ◽  
Con A ◽  
Cellular Activation

Population features of the mouse peritoneal macrophages isolated after stimulation with concanavalin A and thioglycolate

2021 ◽  
pp. 107-115
Author(s):  
E. S. Zubkova ◽  
◽  
K. V. Dergilev ◽  
I. B. Beloglazova ◽  
Yu. D. Molokotina ◽  
...  
Keyword(s):  
Concanavalin A ◽  
Peritoneal Macrophages ◽  
Mouse Peritoneal Macrophages

scholarly journals Phagosome-lysosome interactions in cultured macrophages infected with virulent tubercle bacilli. Reversal of the usual nonfusion pattern and observations on bacterial survival.

1975 ◽  
Vol 142 (1) ◽  
pp. 1-16 ◽  
Author(s):  
J A Armstrong ◽  
P D Hart
Keyword(s):  
Electron Microscopy ◽  
Rabbit Antiserum ◽  
Peritoneal Macrophages ◽  
Bacterial Survival ◽  
Virulent Challenge ◽  
Secondary Lysosomes ◽  
High Level ◽  
Mouse Peritoneal Macrophages ◽  
Specific Rabbit ◽  
Strain H37rv

Tubercle bacilli of the pathogenic human strain H37Rv had previously been shown to multiply, after ingestion by cultured mouse peritoneal macrophages, within phagosomes that tended to remain unfused with secondary lysosomes. Means were sought therefore for promoting experimentally a modification of the host response so as to attain a high level of phagolysosome formation, enabling tests to be made of any effects on the course and outcome of the intracellular infection. This was achieved by exposing viable bacilli to specific rabbit antiserum before their ingestion. Quantitative assessments, using electron microscopy, now showed that a majority of the phagosomes containing intact bacilli had fused with ferritin-labeled lysosomes, and frequently the fusion was massive. Bacterial viability studies established that the serum pretreatment was not itsel bactericidal. In the course of progressive infections with strain H37Rv, monitored by counts both of viable bacterial units and of intracellular acid-fast organisms, no appreciable difference was found between the intracellular growth rates of control and antiserum-treated bacilli. Concurrent electron microscopy showed that bacilli could remain intact and multiply both in phaagolysosomes and in unfused phagosomes, ruling out the possibility of selective growth of antiserum-pretreated bacilli within the minority of phagosomes that remained unfused. It was concluded that "turning on" phagosome-lysosome fusion in normal macrophages did not influence the outcome of infection with virulent M. tuberculosis; lysosome contents manifestly failed to exercise an antibacterial effect on this organism. Nevertheless, the possibility remains that the lysosomes of specific immune macrophages have antituberculous potentiality. In that case the experimental "turning on or off" of fusion could be a decisive factor in the outcome of a virulent challenge. Should it not be, the antibacterial capabilities of immune cells would need to be ascribed to factors other than lysosomal attack, the latter being essentially for disposal of the dead organisms.

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