Characterization of the IgE Fc receptors on monocytes and macrophages.

Subpopulations of human monocytes (15%) and alveolar macrophages (AM phi, 8%) and rat and mouse AM phi (89%) and peritoneal M phi (57%) bear Fc receptors for IgE (Fc epsilon R) as shown by IgE-specific rosette formation. Cells from M phi-like cell lines of human, rat, and mouse origins also express Fc epsilon R. Monomeric IgE binds to Fc epsilon R on M phi with an equilibrium association constant Ka congruent to 10(7) M-1. The Fc epsilon R on human monocytes and M phi are antigenically similar to Fc epsilon R on lymphocytes but differ from Fc epsilon R on basophilic granulocytes. The Fc epsilon R on human and mouse M phi promote phagocytosis and lysis of IgE-coated erythrocytes. Patients with active IgE-mediated allergic diseases have elevated percentages of Fc epsilon R(+) monocytes (56%) that show allergic increased lytic activity against IgE-coated erythrocytes as compared to monocytes from normal humans. M phi from rats infested with Nippostrongylus brasiliensis parasites express more Fc epsilon R than normal M phi. The data indicate that Fc epsilon R expressed on M phi differ from those on mast cells and basophils, increase in number during IgE immune responses, and are likely to play an important role in the host's defense against parasites and in the pathogenesis of allergic diseases.

Characterization of Fc receptors for IgE on human alveolar macrophages.

Human alveolar macrophages (aM phi) isolated from lung lavages performed during bronchoscopy and after surgical removal of pulmonary lobes were analysed for Fc receptors for IgE (Fc epsilon R) and IgG (Fc gamma R) by rosette assays. A mean+/-s.d. of 8.0+/-2.6% of aM phi formed rosettes with fixed ox erythrocytes coated with an IgE myeloma protein (Eo'-IgE). The Eo'-IgE rosettes were inhibited by two IgE myeloma proteins and by IgE Fc fragments but not by myeloma proteins of the other Ig classes or by IgE denatured by heating or reduction and alkylation. Fresh ox erythrocytes sensitized with rabbit IgG antibodies (EoA) formed rosettes with 64.1+/-20.3% of the aM phi. Peripheral blood monocytes formed 10.6+/-1.2% Eo'-IgE and 90.2+/-6.0% EoA rosettes. Incubation of the aM phi with a goat antiserum to human lymphocyte Fc epsilon R inhibited Eo'-IgE rosette formation on aM phi by 80% but did not affect the percentage of EoA rosettes. The antiserum also inhibited Eo'-IgE rosettes formed by peripheral blood monocytes and cultured macrophage-like U937 cells but not those formed by basophilic granulocytes obtained from a patient with chronic myelogenous leukaemia. There was no relationship between age, sex, diagnosis or smoking history of the patients and the percentage of aM phi forming Eo'-IgE rosettes. These studies demonstrate that a subpopulation of human aM phi bear Fc epsilon R that share antigenic determinants with Fc epsilon R on lymphocytes and monocytes. Fc epsilon R(+) aM phi may play an important role in allergic and inflammatory pulmonary diseases by inducing the release of mediators of inflammation after interaction with IgE immune complexes.

Comparison of the Fc receptors for IgE on human lymphocytes and monocytes.

Fc receptors for IgE (Fc epsilon R) on human peripheral blood lymphocytes and monocytes and cultured lymphoblastoid and macrophage-like cell lines were compared with respect to: 1) binding affinity for radiolabeled IgE, 2) inhibition of IgE-specific rosette formation and inhibition of binding of radiolabeled IgE by an antiserum raised against Fc epsilon R isolated from a lymphoblastoid cell line, and 3) m.w. of radiolabeled cell surface proteins precipitated with the anti-Fc epsilon R serum. Scatchard analysis of 125I-IgE binding to lymphocytes, monocytes, and their corresponding cell lines showed biphasic binding curves with all cell types, from which 2 binding affinities were calculated to be KA = 6.2 +/- 1.1 and 2.0 +/- 0.5 x 10(7) M-1. The anti-Fc epsilon R serum inhibited both IgE rosette formation and binding of radiolabeled IgE by lymphocytes and monocytes but did not inhibit IgE rosettes formed by basophils. The inhibitory activity of the anti-Fc epsilon R serum could be absorbed with Fc epsilon R(+) but not with Fc epsilon R(-) cell lines. The anti-Fc epsilon R serum precipitated 2 peptides having m.w. of approximately 47,000 and 23,000 daltons from lysates of both cell surface-labeled lymphocyte and macrophage cell lines. These data indicate that Fc epsilon R on normal lymphocytes and monocytes, as well as on cultured lymphoblastoid and macrophage-like cells, are related structurally, since they share antigenic determinants, bind IgE with a similar affinity, and have similar m.w. However, they differ in all 3 parameters from Fc epsilon R on basophilic granulocytes.

Increased peripheral blood monocytes with Fc receptors for IgE in patients with severe allergic disorders.

Peripheral blood monocytes, defined as latex bead-ingesting mononuclear cells, from 15 healthy nonallergic donors and 22 patients with allergic disorders were analyzed for Fc receptors for IgE (Fc epsilon) by a rosette assay employing ox erythrocytes coated with IgE. The patients were divided into 3 groups. Group I: 12 patients with mild to moderate atopic disease and serum IgE levels up to 2300 IU/ml. Group II: 6 patients with severe generalized atopic dermatitis, allergic rhinitis, and asthma, of which 5 had IgE serum levels of 8000 to 77,500 IU/ml. Group III: 4 severely atopic patients with IgE levels greater than 10,000 IU/ml and receiving oral corticosteroids. The numbers of monocytes were similar in healthy donors and patients. In contrast, severely atopic patients (Group II) had significantly more (p less than 0.01) Fc epsilon + monocytes (107 +/- 42/mm3) than healthy donors (20 +/- 14/mm3) or patients of Group I (31 +/- 14/mm3). Patients of Group III had significantly fewer (p less than 0.05) Fc epsilon + monocytes (12 +/- 16/mm3) than controls and patients of Groups I and II. We conclude that patients with severe allergic disorders have a significant increase of peripheral blood monocytes with Fc receptors for IgE, which suggests that these cells may participate in the pathophysiology of atopic disease.

Increased IgE-dependent cytotoxicity by blood mononuclear cells of allergic patients.

Peripheral blood mononuclear cells from 14 healthy donors and 22 allergic patients were incubated with 51Cr-labelled chicken erythrocytes coated with an IgE myeloma protein or rabbit IgG antibodies. Mononuclear cells from patients with severe atopic disorders released a significantly greater percentage of 51Cr (P less than 0.001) from IgE-coated target cells than mononuclear cells from healthy controls, patients with mild atopic disease, or patients with severe atopic disease taking oral prednisone. Specific 51Cr-release from IgE-coated target cells was directly correlated to the percentage of monocytes (latex-ingesting cells) with Fc receptors for IgE (r = 0.87, P less than 0.01) as detected by a rosette assay employing ox erythrocytes coated with IgE. Mononuclear cells from patients and normals released similar amounts of 51Cr from IgG-sensitized target cells. Depletion of monocytes from mononuclear cell preparations from two severe atopic patients decreased 51Cr-release from IgE-coated target cells to levels seen in healthy donors or patients with mild allergic disease. These results demonstrate that mononuclear cells from severely allergic patients have a significantly increased cytotoxicity toward IgE-coated targets coated target cells and that this cytotoxicity correlates highly with the percentage of monocytes with Fc receptors for IgE in these mononuclear preparations.

The mononuclear cell in human blood which mediates antibody-dependent cellular cytotoxicity to virus-infected target cells. II. Identification as a K cell.

Studies were carried out to determine whether the mononuclear cell in human blood which mediates antibody-dependent cellular cytotoxicity (ADCC) to herpes simplex virus (HSV)-infected target cells has surface Fc receptors which participate in the reaction. The F (ab')2 fragment of human IgG antibody was inactive both in ADCC and in complement-mediated cytolysis, but retained the capacity to neutralize infectious virus, to agglutinate erythrocytes coated with viral antigens, and to bind to the surface of virus-infected cells. Treatment of sensitized virus-infected target cells with staphylococcus protein A, which has affinity for the Fc epitope of IgG, strongly reduced their susceptibility to lysis by ADCC in a dose-dependent relationship. These findings indicate that the Fc portion of IgG antibody to the virus is necessary for cytotoxicity. Treatment of blood mononuclear cells with either heat-aggregated gamma-globulin or HSV immune complexes inhibited effector cell activity. The presence of "third party" cellular immune complexes also strongly inhibited ADCC. Adsorption of mononuclear cells to plastic surfaces coated with soluble third party immune complexes resulted in a significant reduction in effector cell activity. These findings demonstrate that the ADCC effector cell possesses surface Fc receptors which are utilized in the ADCC reaction. The presence of Fc receptors on the surface of the effector cell indicates that it is a K cell rather than a null cell.

The mononuclear cell in human blood which mediates antibody-dependent cellular cytotoxicity to virus-infected target cells. I. Identification of the population of effector cells.

Mononuclear cells (MC) from human blood were fractionated by a variety of physical and immunologic techniques, and the cellular subpopulations generated were assessed for their capacity to lyse herpes simplex virus (HSV)-infected target cells in the presence of IgG antibody to HSV. Latex phagocytosis and surface marker studies were performed in parallel in order to identify the major effector cells by their phagocytic properties and their possession of surface immunoglobulin and receptors for either sheep erythrocytes, C3, or the Fc fragment of IgG. Cytotoxic effector cell activity was unaffected or slightly enhanced after the removal of plastic-adherent or carbonyl iron-adherent MC, indicating that the major effector cell is not a classical monocyte. Similar results were obtained after removal of more than 90% of the T cells by depletion of rosette-forming cells. Likewise, effector cell activity was generally unchanged when more than 95% of the B cells were removed by filtering MC on nylon wool columns. Effector cell function was also found to be normal in three patients with B cell-deficient X-linked agammaglobulinemia. These observations strongly suggest that the effector cells are not T cells or B cells. A 4- to 5-fold enrichment in effector cells, however, was consistently found in a subpopulation, consisting of 5% of the unfractionated MC, that was dramatically enriched both for nonphagocytic cells with only Fc receptor (K cells) and for nonphagocytic cells with no detectable surface markers (null cells). Since, as is demonstrated in the accompanying report, effector surface Fc receptors play a critical role in the mediation of antibody-dependent cellular cytotoxicity directed at HSV-infected target cells, the major mononuclear effector cell in human blood is a K cell.

Antibody-dependent cell-mediated cytotoxicity to target cells infected with type 1 and type 2 herpes simplex virus.

The phenomenon of antibody-dependent cell-mediated cytoxicity (ADCC) has been extended to include target cells acutely infected with herpes simplex type 1 virus (HSV-1) or herpes simplex type 2 virus (HSV-2) in an in vitro system that employs immune human serum and human blood mononuclear cells. The cytotoxic reaction was detectable after 1 hr of incubation and was complete between 4 and 8 hr. The amount of ADCC noted was directly proportional to the logarithm(10) of the effector: target cell ratio (E:T), and ADCC was noted at E:T as low as 1:1. The mononuclear effector cell was present in the blood of both HSV immune and non-immune individuals. The immune serum factor was demonstrated to be an antibody with specificity for HSV membrane antigen(s) and was reactive with target cells infected with either of the two HSV types. The antibody rendered the mononuclear cell cytotoxic by sensitization of the target cell rather than by direct attachment to or "arming" of the mononuclear cell. The physiochemical properties of the antibody as well as its presence in cord blood demonstrated that it is an immunoglobulin on the IgG class.