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The demonstration of cells bearing Fc receptors in the metrial gland of the pregnant rat uterus.

Cells obtained by collagenase treatment of metrial gland tissue from rats of day 12, 13, 14 and 15 of pregnancy were examined for the presence of surface membrane receptors for immunoglobulin (Fc receptors). Using an EA rosetting technique in which sheep red blood cells (SRBC) were sensitized with a rabbit anti-SRBC immunoglobulin preparation, Fc receptors were found on a proportion of the cells. The majority of the granulated metrial gland cells were not included in the rosetting cell population, suggesting that they do not possess the type of Fc receptor detected by this method. A comparison was made between results obtained when cells were counted in suspension and those obtained from cell counts on sections of fixed material. Both methods were found to yield similar results.

Animals

Fc receptor bearing 'hairy cells' of leukaemic reticuloendotheliosis bind soluble antigen--antibody complexes and adhere to immobilized complexes, but fail to mediate antibody dependent cellular cytotoxicity.

The majority of hairy cells from three patients with leukaemic reticuloendotheliosis were adherent cells bearing surface immunoglobulin, complement receptors, and Fc receptors. Highly purified populations of malignant hairy cells, which readily bound soluble antigen-antibody complexes in suspension and were able to adhere to immobilized antigen-antibody complexes, were examined for their ability to mediate ADCC. Two patients with greater than 90% FcR positive cells failed to mediate ADCC. When initially examined, a third patient, with fewer malignant cells, demonstrated a less marked impairment of cytolysis. When reexamined at a later date, this patient had an increased number of hairy cells and concomitantly demonstrated more impaired ADCC effector cell activity. Absorption of surface immunoglobulin bearing cells onto plastic surfaces coated with goat anti-human immunoglobulin resulted in a complete depletion of FcR, sIg positive, hairy cells. The remaining nonadherent fraction, containing 5.5% FcR positive, sIg negative cells, was able to mediate ADCC as effectively as the normal controls. These results indicate that although FcR bearing hairy cells readily bind soluble antigen-antibody complexes and adhere to immobilized complexes, they were unable to mediate ADCC.

Adult

Electron microscopy of Fc receptors on human lymphocytes.

The membrane receptor for Fc portions of IgG (FcR) was localized on the cell surface of humans lymphocytes by electron microscopy. The electron microscopic markers for FcR were soluble ferritin 7S anti-ferritin immune complexes prepared in forty times antigen excess than needed at equivalence. Fc receptors on the lymphocytes labelled at 0 degree in the presence of sodium azide were seen as discontinuous patches on the cell surface. In control experiments, no labelling was observed, which included lymphocytes treated with ferritin only or with F(ab')2 immune complexes as well as glutaraldehyde-fixed lymphocytes treated with 7S anti-ferritin immune complexes. The findings are discussed with relation to the widely accepted membrane fluidity model.

Antigen-Antibody Complex

Fc receptors on mouse effector cells mediating natural cytotoxicity against tumor cells.

Mouse effector cells mediating natural cytotoxicity against tumor cells have been previously thought to be lymphocytes that lack any detectable cell surface markers. The present study presents evidence for receptors for the Fc portion of IgG on these cells. By adsorption of cytotoxic spleen cells on monolayers of sheep erythrocytes (E) plus IgG antibodies to sheep erythrocytes (EA), 50 to 96% of the total cytotoxic reactivity could be removed. Parallel adsorption of cells on E monolayers or on EA monolayers coated with protein A, to block the Fc portion of IgG, resulted in little or no depletion of cytotoxic activity. The presence of Fc receptors on the NK cells was confirmed by combining EA rosette formation with velocity sedimentation at unit gravity. Peak cytotoxicity occurred at the same sedimentation velocity as the peak of Fc-positive cells. After EA rosette formation, there was a shift to a higher sedimentation velocity in the Fc-positive cells and in the natural cytotoxic activity. The increase in sedimentation velocity of NK activity that was observed in these experiments indicated that most of the cells had only bound a small number (three or four) of antibody-coated erythrocytes. Together, these data indicate that cells with Fc receptors account for most of the total lytic activity of normal mouse spleen cells.

Animals

Possible role of Fc receptors on cells infected and transformed by herpesvirus: escape from immune cytolysis.

Receptors for the Fc portion of nonimmune immunoglobulin G were demonstrated on B103 rat brain neuroma cells infected with herpes simplex virus type 1 (HSV-1) KOS by a radioimmunoassay using 125I-labeled heat-aggregated Fc fragments. Immune F(ab')2 fragments specific for HSV antigens competed efficiently for Fc binding sites, suggesting that the binding of Fc fragments to infected cells is specific for viral cell-surface antigens. It has been suggested that the binding of immune complexes to Fc receptors on the surfaces of tumor cells in vivo plays a role in protecting these cells from immune destruction. In vitro evidence is presented for the ability of aggregated immunoglobulin G molecules bound to cell-surface Fc receptors to protect both HSV-infected and HSV-transformed cells against complement-dependent and cell-mediated immune lysis.

Animals

Separation and functional analysis of subpopulations of lymphocytes bearing complement and Fc receptors.

A highly versatile procedure is described in this review which can be used to separate and obtain in pure form subpopulations of lymphoid cells which express different cell surface structures. The method is based on the observation that when rosetting and non-rosetting leukocytes are centrifuged on a cushion of Isopaque/Ficoll, the rosetting leukocytes and red cells sink whereas the non-rosetting leukocytes float. Thus, any subpopulation of leukocytes can be separated providing they can be identified by rosetting. The earlier sections of this review describe the method, its efficiency of separation and its advantages compared with other fractionation procedures. Subsequent sections describe experiments in which the procedure was specifically applied to separating Fc receptor (Fc+) and complement receptor (CR+) lymphocytes. On the basis of these two receptors it was possible to subdivide T and B lymphocytes into distinct subpopulations. Four subclasses of B lymphocytes were identified in mouse spleen (Fc+CR+,Fc+CR-,Fc-CR+ and Fc-CR-) and two subclasses of T cells were also detected (Fc+ and Fc-). The functional relevance of these subpopulations of lymphocytes was examined. It was found that in all cases examined, antigens could successfully activate CR+ B cells to produce antibody. However, only polymeric antigens, whether T-dependent or T-independent, were capable of triggering CR- B cells to synthesize antibody. Furthermore, preliminary experiments suggest that Fc receptors are present on functional B cells and helper T cells but are not expressed on cytotoxic T cells. On the basis of these results it is proposed that complement receptors on B lymphocytes provide an additional binding site which stabilizes the union between the antigen-specific receptors and soluble antigen. In contrast, due to their multi-determinant nature, polymeric antigens can avidly bind to B cells without involvement of the complement receptors. The possibility of Fc receptors playing a similar role in stabilizing the interaction of antigen with specific receptors on lymphocytes, particularly on T helper cells, is also discussed.

Animals

Multiple FC receptors in the human placenta.

The presence of Fc receptors on endothelium and trophoblastic tissue in cryostat sections of full-term human placentae has been verified. Fc receptor activity on stem vessel endothelium can be manifest both by immunofluoresence with heat-aggregated IgG and by haemadsorption with antibody-sensitized erythrocytes. However, Fc receptor activity on trophoblastic tissue is manifest only in the haemadsorption reaction, and not in the immunofluorescence reaction. The haemadsorption and immunofluorescence were both independent of temperature, and were completely abolished by formaldehyde fixation of the tissue sections. Both reactions were expressed on first-trimester placental tissue, although more limited and diffuse. In terms of their anatomical location and specificity characteristics the Fc receptors in human placentae may have a biological significance.

Endothelium

Effects of cholera exotoxin on Fc receptor activity of lymphoid cells and mononuclear phagocytes.

The effect of cholera exotoxin and aminophylline on Fc receptors in a murine lymphoid-cell line and in rabbit pulmonary alveolar macrophages has been investigated. Although both agents elevated intracellular cyclic AMP levels in macrophages and lymphoid cells, the effects on Fc receptor expression were distinct. Cholera toxin at 10 microng/ml reversibly inhibited Fc-receptor activity in the murine lymphoid cell line. In contrast, cholera toxin at 10 microng/ml or 0-01 microng/ml was ineffective in altering pulmonary alveolar macrophage receptor expression. Fc receptor activity on the macrophage was reduced by 20-30 per cent following incubation with aminophylline, (10(-3)M) from 0-6 h. There was no direct correlation between Fc-receptor activity and cyclic AMP levels in the cells studied. The differential susceptibility of these lymphoid and phagocytic cell populations to cholera toxin and also toward aminophylline suggests that there may be fundamental differences in topography on the membrane surface, or in the intracellular regulation of Fc receptors between lymphoid and phagocytic cells.

Aminophylline

The effects of pharmacologic agents on immune-complex-induced redistribution of B-lymphocyte Fc receptors.

The effects of pharmacologic agents on the immune-complex-induced redistribution of B-lymphocyte Fc receptors (and, as a control, the anti-Ig induced redistribution of surface Ig) were examined. Immune-complex-induced capping of B-cell Fc receptors was moderately to markedly inhibited by the combination of colchicine and cytochalasin B, the Ca++ ionophore A23187, and the local anaesthetic lidocaine but was only slightly inhibited by cytochalasin B alone and was not inhibited by colchicine alone. Inhibition of capping was not due to the inhibition of binding of immune complexes to the B-lymphocytes or to decreased cell viability since these effects were absent. Preformed immune complex-Fc receptor caps were disrupted by A23187, lidocaine, and the combination of colchicine and cytochalasin B, but not by either colchicine or cytochalasin B alone. The effects of the pharmacologic agents were similar for Fc receptors and surface Ig in all cases. These results suggest that ligand bound Fc receptors are affected by cytoskeletal structures and that the ligand-induced redistribution of two distinct B lymphocyte surface receptors (fc receptors and surface Ig) occurs by similar or identical mechanisms.

Animals

Characterization of lymphoblast Fc receptor expression in acute lymphoblastic leukemia.

Lymphoblasts from 18 patients with untreated acute lymphoblastic leukemia were investigated for the presence of conventional cell surface markers (spontaneous sheep red blood cell rosette formation, complement receptors, and surface immunoglobulin). The expression of Fc receptors for both IgG and IgM was investigated using indicator bovine erythrocytes coated with rabbit anti-BRBC (IgG and LgM fractions). The leukemic cells of all patients in this tudy expressed Fc receptors for both IgC and LgM. In contrast with previous reports, null (non-T non-B) lymphoblasts as well as T lymphoblasts demonstrated Fc-IgG and Fc-IgM receptors. However, these two immunologic subclasses of leukemic cells demonstrated significantly different patterns of Fc receptor distribution. These data suggest that expression of Fc receptors is an early event in cellular differentiation in this lymphoid malignancy.

Adolescent

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

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.

Animals

Fc-receptors, Ia-antigens, and immunoglobulin on normal and activated mouse T lymphocytes.

Using antibody coated bovine erythrocytes we were unable to demonstrate Fc-receptors on either thymus cells or T cells prepared from lymph node cell suspensions by anti-Ig column filtration. However, if parental thymus or lymph node T cells were transferred to X-irradiated F1 hybrids, activated donor T cells recovered from the recipient's spleen (ATC-spleen) were shown to express Fc-receptors. Fc-receptors were also demonstrable on ATC-spleen prepared between strain combinations differing at the M-locus. In marked contrast, Fc-receptors were not detected on ATC recovered from thoracic duct lymph (T.TDL). This applied to (a) H-2-activated T.TDL derived from normal thymus cells, (b) H-2-activated T.TDL derived from thymus cells depleted of B cells, and (c) M-locus-activated T.TDL. Of these three populations, surface Ig (of B cell origin) was detected on a large proportion of the first but not on the second and third populations. Thus, the failure to detect Fc-receptors on any of these populations could not be attributed to blocking by adsorbed surface Ig. In addition, various T-cell populations were examined by a microcytotoxicity assay for the presence of cell surface Ia-antigens. 5--10% of the thymus cells, 20--30% of cortisone-resistant thymus cells, 60--70% of lymph node cells, and 60--80% of ATC-spleen and T.TDL showed Ia.

Animals

Comparison of various tests for Fc receptors on different human lymphocyte sub populations.

Six different immune complex test systems for the detection of IgG Fc receptors were applied to the study of various human lymphocyte populations. The extent of binding varied widely according to the system and the cell type employed. Two systems bound preferentially to a high proportion of B lymphocytes from peripheral blood or tonsils, one of which bound with only a very few T cells. In contrast, four other test systems which bound well with the Fc receptors on T lymphocytes gave weaker reactions with Fc receptors on B cells. The reactivity of Fc receptors on null or third population lymphocytes was similar to that of the Fc-positive T cells. Pronase digestion experiments showed a graded selective loss of reactivity with the different Fc reagents. No one system was optimal for all of the lymphocyte populations, although aggregated IgG exhibited the broadest spectrum of reactivity. A pronounced effect of temperature was evident on the binding reactions, and native IgG showed strong binding at 4 degrees C, particularly to the Fc receptors on T cells.

Antigen-Antibody Complex

The characterization and functional significance of plasma membrane Fc Receptors.

The importance of various components of the plasma membrane in the generation of the immune response has long been recognized. The identification and characterization of functional membrane receptors are essential for an understanding of the molecular basis of the immune response. The Fc receptor can be operationally defined as a site on the plasma membrane which is capable of binding the Fc portion of IgG. The Fc receptor has been reported on both lymphoid and nonlymphoid cell types. The functional significance of the receptor on these cell populations, as well as the biochemistry of the receptor, is not well established and is currently an important area of investigation. In this review, the Fc receptor will be discussed, primarily in human and murine lymphoid and macrophage cell populations, as a model of membrane immunoreceptors. The review will be divided into four sections. In the first section, consideration will be given to the binding assays used to study Fc receptor-ligand interactions and the class and subclass specificity of the receptor. The possibility of receptor-receptor interaction and quantitation of receptor sites will be discussed. The second phase of the review will be concerned with the fractionation and biochemistry of this receptor system, emphasizing the differences reported in receptor composition from different cell sources. In the third section of the review, the biologic significance of the receptor will be examined. The importance of the Fc receptor in opsonization and phagocytosis by mononuclear phagogytes, B-cell activation, antibody-dependent cell cytotoxicity, and the possible relationship of Fc receptors to the histocompatibility-linked immune response genes will be discussed. Finally, in the fourth section, the possible mechanisms of receptor regulation will be considered. Cell fusion experiments which have been performed and the expression of receptor activity on the hybrid cell clones will be discussed. Receptor regulation at the physiologic level will include the effects of cyclic nucleotide levels and cell cycle on receptor expression. It is the intent of this review to provide a detailed analysis of the Fc receptor and to emphasize its importance in microbiology, cell biology, and immunology.

Animals