scholarly journals A new Fc receptor on mouse macrophages binding IgG3.

1981 ◽  
Vol 153 (3) ◽  
pp. 514-519 ◽  
Author(s):  
B Diamond ◽  
D E Yelton
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Cytochalasin B ◽  
Fc Receptors ◽  
Peritoneal Macrophages ◽  
Fc Receptor ◽  
Igg Subclasses ◽  
Macrophage Cell ◽  
Sheep Erythrocytes ◽  
Mouse Macrophages

Monoclonal antibodies to sheep erythrocytes (SRBC) have proved useful in identifying two Fc receptors on mouse macrophages, one for IgG2a, and one for IgG1 and IgG2b. We have used monoclonal IgG3 anti-SRBC to identify a third Fc receptor on mouse macrophages which binds IgG3 uniquely. This receptor is present on primary resident and thioglycolate-induced peritoneal macrophages and on some macrophage cell lines. The binding of IgG3-coated SRBC is inhibited by aggregated byt not monomeric IgG3, and not by IgG1, IgG2a, and IgG2b aggregates. It is unaffected by treating the macrophages with trypsin or cytochalasin B and occurs at both 4 degrees and 37 degrees C. IgG3, like all other IgG subclasses, mediates phagocytosis. We have also generated a variant macrophage line which bears the receptors for IgG1 and IgG2b and for IgG2a, but not for IgG3.

scholarly journals Structural evidence for distinct IgG subclass-specific Fc receptors on mouse peritoneal macrophages

1980 ◽  
Vol 152 (5) ◽  
pp. 1147-1161 ◽  
Author(s):  
BC Lane ◽  
J Kan-Mitchell ◽  
MS Mitchell ◽  
SM Cooper
Keyword(s):  
Binding Proteins ◽  
Sodium Dodecyl ◽  
Fc Receptors ◽  
Peritoneal Macrophages ◽  
Fc Receptor ◽  
Rabbit Igg ◽  
Mouse Macrophages ◽  
Specific Proteins ◽  
Sepharose 4B ◽  
Mouse Peritoneal Macrophages

Membrane proteins which selectively bind to the Fc portion of IgG were identified in the Nonidet P-40 extracts of radiolabeled thioglycollate- elicited mouse peritoneal macrophages. Affinity columns of various IgG preparations coupled to Sepharose 4B were used to absorb the Fc-binding proteins. Analysis of the acetic acid or sodium dodecyl sulfate (SDS) eluates from aggregated human IgG or antigen-complexed rabbit IgG columns revealed two Fc(gamma)/-specific proteins with apparent 67,000 and 52,000 mol wt. These proteins were not detected in acid or SDS eluates from F(ab')(2) columns or in eluates from IgG column, over which were passed lysates of Fc receptor-negative cells. With the use of affinity columns that contained aggregated mouse myeloma proteins of different IgG subclasses, we found that the 67,000-dahon protein selectively binds to IgG2a, whereas the 52,000-dalton protein binds to IgG1 and IgG2b. Neither protein was found in SDS eluates from IgG3 columns. Trypsin treatment of the macrophages before detergent lysis removed the 67,000-dalton protein, although it leaves intact the 52,000-dalton protein. These results provide structural confirmation for the existence of separate Fc receptors on mouse macrophages and indicate that the two Fc-binding proteins identified in this study represent all or part of the trypsin- sensitive Fc receptor which binds IgG2a and the trypsin-resistant Fc receptor which binds IgG2b and IgG1.

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 Studies of the macrophage complement receptor. Alteration of receptor function upon macrophage activation.

1975 ◽  
Vol 141 (6) ◽  
pp. 1278-1290 ◽  
Author(s):  
C Bianco ◽  
F M Griffin ◽  
S C Silverstein
Keyword(s):  
Macrophage Activation ◽  
Quantitative Difference ◽  
Qualitative Difference ◽  
Fc Receptors ◽  
Significant Degree ◽  
Peritoneal Macrophages ◽  
Fc Receptor ◽  
Complement Receptor ◽  
Complement Receptors ◽  
Activated Macrophages

We have examined the roles of Fc receptors and complement receptors in mediating the interaction of sensitized sheep erythrocytes (E) with activated and with nonactivated mouse peritoneal macrophages. Both activated and nonactivated macrophages ingest IgG-coated erythrocytes [E(IgG)]; activated cells intest 1.5-2 times as man E(IgG) as do nonactivated macrophages. Thus, there is a quantitative difference in Fc receptor-mediated ingestion between activated and nonactivated macrophages. There is, however, a qualitative difference in function of complement receptors of activated and nonactivated macrophages. Nonactivated macrophages avidly bind complement-coated E [E(IgM)Ia1, but do not ingest them to a significant degree. Activated macrophages, on the other hand, bind and ingest E(IgM)C. The possibility of Fc receptor participation in mediating ingestion of E(IgM)C by activated macrophages was eliminated by blocking Fc receptors with an antimacrophage IgG fraction. Activated macrophages treated with antimacrophage IgG did not ingest E(igG) but did ingest both E(IgM)C AND E(IgM)C. Nonactivated macrophages treated with antimacrophage IgG did not interact at all with E(IgG). These cells bound, but did not ingest, E(IgM)C and E(IgM)C. Complement receptor-mediated ingestion is a marker for macrophage activation and may be physiologically important in the elimination of complement-coated particles.

scholarly journals Loss of Fc receptor activity after culture of human monocytes on surface-bound immune complexes. Mediation by cyclic nucleotides.

1980 ◽  
Vol 151 (1) ◽  
pp. 32-44 ◽  
Author(s):  
C G Ragsdale ◽  
W P Arend
Keyword(s):  
Immune Complexes ◽  
Cytochalasin B ◽  
Fc Receptors ◽  
Phosphodiesterase Inhibitor ◽  
Fc Receptor ◽  
Human Monocytes ◽  
Partial Loss ◽  
Mediated Effects ◽  
Receptor Number ◽  
Partial Reversal

Human monocytes cultured on surface-bound immune complexes exhibited a loss of ability to form rosettes with IgG-sensitized sheep erythrocytes (EA). This loss was not a result of inhibition of Fc receptors by solubilized complexes nor of release of soluble factors by the cells. Loss of EA rosetting was not prevented by culture of monocytes at 4 degrees C, or by treatment with colchicine, cytochalasin B, or local anethetic agents. These results suggested that the loss was not secondary to capping or interiorization of Fc receptors. The results of other studies indicated that the Fc receptors were not damaged by lysosomal enzymes or oxygen radicals. Maintenance of EA rosetting ability of monocytes cultured on surface-bound immune complexes was seen after a 3-h preincubation of the cells in 100 mM 2-deoxy-D-glucose (2dG). A similar preincubation in ATP or in 8-bromoadenosine 3':5'-cyclic monophosphoric acid plus the phosphodiesterase inhibitor methyl isobutyl xanthine led to a partial loss of EA rosetting of cells on plain fibrin and to a partial reversal of the effects of 2dG seen with cells on complexes. We conclude that EA rosetting of monocytes cultured on surface-bound immune complexes is reduced by cyclic nucleotide-mediated effects on Fc receptor number or function.

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.

scholarly journals Colocalization of F-actin and talin during Fc receptor-mediated phagocytosis in mouse macrophages.

1990 ◽  
Vol 172 (6) ◽  
pp. 1853-1856 ◽  
Author(s):  
S Greenberg ◽  
K Burridge ◽  
S C Silverstein
Keyword(s):  
Peritoneal Macrophages ◽  
Fc Receptor ◽  
Early Stages ◽  
Mouse Macrophages ◽  
J774 Cells ◽  
Particle Ingestion ◽  
Coordinate Changes ◽  
Mouse Peritoneal Macrophages ◽  
Cytoplasmic Location

We have studied the distribution of talin in J774 cells and mouse peritoneal macrophages undergoing Fc receptor-mediated phagocytosis. At early stages of phagocytosis, talin accumulates in the cells' cortical cytoplasm adjacent to the forming phagosome and extends into pseudopods that are encircling the particle. Talin colocalizes with F-actin at these sites. After particle ingestion is completed, F-actin and talin are no longer concentrated adjacent to phagosomes. Thus, talin and F-actin undergo dynamic and coordinate changes in their cytoplasmic location during Fc receptor-mediated phagocytosis.

scholarly journals Internalization and degradation of macrophage Fc receptors bound to polyvalent immune complexes.

1984 ◽  
Vol 98 (4) ◽  
pp. 1170-1177 ◽  
Author(s):  
I Mellman ◽  
H Plutner
Keyword(s):  
Immune Complexes ◽  
Fc Receptors ◽  
Fc Receptor ◽  
Surface Receptors ◽  
Molar Ratios ◽  
Mouse Macrophages ◽  
Immune Precipitation ◽  
Specificity Of Binding ◽  
Receptor Antibodies ◽  
Receptor Turnover

We have studied the Fc receptor-mediated pinocytosis of immunoglobulin G (IgG)-containing immune complexes by mouse macrophages. IgG complexes were formed from affinity-purified rabbit dinitrophenyl IgG and dinitrophenyl modified BSA at molar ratios of 2.5-10:1. Both the specificity of binding and the fate of internalized receptors were analyzed using monoclonal and polyclonal anti-Fc receptor antibodies. Based on the susceptibility of surface-bound ligand to release by proteolysis, we have found that at 37 degrees C, 125I-labeled IgG complexes were rapidly internalized (t1/2 less than 2 min) and delivered to lysosomes; acid-soluble 125I was detectable in the growth medium within 5-10 min of uptake. However, kinetic evidence indicated that Fc receptors were not efficiently re-used for multiple rounds of ligand uptake. Instead, macrophages that were exposed continuously to saturating concentrations of IgG complexes exhibited a selective and largely irreversible removal of Fc receptors from the plasma membrane. This loss of surface receptors correlated with an increased rate of receptor turnover, determined by immune precipitation of Fc receptors from 125I-labeled macrophages. Thus, in contrast to the results obtained in the accompanying paper (I. Mellman, H. Plutner, and P. Ukkonen, 1984, J. Cell Biol. 98:1163-1169) using a monovalent ligand, these data indicate that the interaction of Fc receptors with polyvalent complexes leads to the degradation of both ligand and receptor following their delivery to lysosomes.

scholarly journals Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors.

1979 ◽  
Vol 150 (3) ◽  
pp. 580-596 ◽  
Author(s):  
J C Unkeless
Keyword(s):  
Monoclonal Antibody ◽  
Cell Lines ◽  
Fc Receptors ◽  
Inhibitory Effect ◽  
Fc Receptor ◽  
Antigenic Relationship ◽  
Mouse Macrophage ◽  
Spleen Cells ◽  
Binding Studies ◽  
Fab Fragment

To investigate the antigenic relationship between the macrophage and lymphocyte Fc receptors (FcR), a monoclonal antibody capable of blocking mouse macrophage Fc receptors was selected. Hybrids were formed by fusing the P3U1 mouse myeloma and spleen cells from a rat immunized with the mouse macrophage-like cell lines J774 and P388D1. The Fab fragment of the monoclonal IgG secreted by clone 2.4G2, inhibited the trypsin-resistant Fc receptor II (FcRII), which is specific for immune aggregates of mouse IgG1 and IgG2b, but had no inhibitory effect on the trypsin-sensitive Fc receptor I (FcRI), which binds monomeric IgG2a and erythrocytes coated with IgG2a. Thus, the monoclonal 2.4G2 IgG appeared to be specific for macrophage FcRII. Further evidence that the 2.4G2 IgG was directed against FcRII came from binding studies of the monoclonal antibody to J774 cells and a series of independently isolated variants which do not express FcRII. These variants of J774 bound 5% as much of the monoclonal antibody as the parent line, which bound 600,000 molecules of 2.4G2 IgG per cell. The antigenic relatedness of mouse lymphocyte FcR to mouse macrophage FcRII was demonstrated by the binding of 2.4G2 IgG to FcR-bearing lymphoid cell lines and the inhibition of the lymphocyte FcR by the monoclonal antibody. Preincubation of spleen cells and peritioneal cells with 2.3G2 IgG likewise inhibited rosette formation with ox erythrocytes coated with rabbit IgG. The ability of the hybridoma IgG to inhibit mouse FcRII was independent of the major histocompatibility complex. The 2.4G2 IgG antigenic determinant was not present on rat, guinea pig, rabbit, or human FcR-bearing cells.

scholarly journals Effect of cytochalasin B on the adhesion of mouse peritoneal macrophages.

1976 ◽  
Vol 69 (2) ◽  
pp. 407-414 ◽  
Author(s):  
T G Helantjaris ◽  
P S Lombardi ◽  
L A Glasgow
Keyword(s):  
Acid Phosphatase ◽  
Normal Mouse ◽  
Cytochalasin B ◽  
Functional Relationship ◽  
Peritoneal Macrophages ◽  
Phagocytic Cells ◽  
Mouse Macrophages ◽  
Macrophage Adhesion ◽  
Glass Surfaces ◽  
Mouse Peritoneal Macrophages

The adhesion of normal mouse macrophages to glass surfaces was reduced by nontoxic levels (1-50 mug/ml) of cytochalasin B in combination with a centrifugal force (1,000-8,000 g). Macrophages nonspecifically activated by Corynebacterium acnes were also detached by this treatment, but less effectively. The effects of cytochalasin B treatment on these cells were shown to be reversible. After detachment, the cells reattached to glass, appeared morphologically normal, and behaved like untreated cells as judged by adhesion, acid phosphatase levels, and phagocytosis. The effect of cytochalasin B on several parameters of phagocytosis by normal macrophages was also examined. The results demonstrate that cytochalasin B can be used to detach macrophages from surfaces and suggest a functional relationship between phagocytosis and macrophage adhesion to surfaces. Furthermore, the effect of cytochalasin B on adhesion of phagocytic cells provides a probe for further investigation of the adhesion of cells to surfaces.

Sign in / Sign up

Export Citation Format

Share Document