Production of IgE-potentiating factor in man by T cell lines bearing Fc receptors for IgE

The regulation of human IgE production in vitro by soluble T cell factors was examined. T cells were isolated from the peripheral blood of 2 patients with the hyper-IgE syndrome on the basis of their expression of Fc receptors for human IgE (Fc epsilon R). The T cells were incubated with human myeloma IgE (10 micrograms/ml), washed, reacted with immunosorbent-purified goat anti-human IgE conjugated with fluorescein isothiocyanate, and then separated into Fc epsilon R+ and Fc epsilon R- T cells on the fluorescence-activated cell sorter. Fc epsilon R+ T cells and Fc epsilon R- T cells were propagated in culture using supernatants of phytohemagglutinin-stimulated peripheral blood mononuclear cells (PBMC) and irradiated autologous PBMC. Supernatants of Fc epsilon R+ T cell lines but not of Fc epsilon R- T cell lines selectively enhanced IgE synthesis in cultures of B cells obtained from patients with allergic rhinitis but not from normal nonallergic subjects. The surface phenotype of the Fc epsilon R+ T cell line was predominantly T3+, T4+, Ia+ with few (15%) T8+ cells. Two T cell clones were grown from the Fc epsilon R+ T cell line by limiting dilution (0.3 cells/well). These clones possessed the T4+ helper/inducer phenotype and secreted IgE-enhancing factor(s). The IgE-enhancing factor(s) which had affinity for insolubilized human IgE was sensitive to treatment with trypsin and neuraminidase, and had as its target an IgE-bearing B cell. These results suggest that a subset of human T cells bearing an Fc epsilon R secretes an IgE-binding glycoprotein which selectively enhances IgE synthesis by IgE-bearing B cells.     

Regulation of in vivo expression of Fc receptors for IgE (Fc epsilon R) on murine lymphocytes. I. Detection of Fc epsilon R+ lymphocytes by flow cytometry

Homologous monomeric IgE was employed in a flow cytometric assay for the detection of IgE Fc receptors (Fc epsilon R) on mouse lymphocytes. The expression of Fc epsilon R in normal BALB/c mice was detected on splenic and circulating lymphocytes, but not on bone marrow cells. The Fc epsilon R expression was observed in B cells with B220, surface IgM, and IgD, but not in T cells. Infection of mice with Nippostrongylus brasiliensis resulted in a marked increase in the expression of Fc epsilon R on splenic B cells. T cells, however, did not express Fc epsilon R even after N. brasiliensis infection. On the other hand, the Fc epsilon R expression on normal B cells decreased after a simple incubation at 37 degrees C for 24 h, while in the presence of IgE this decrease was inhibited. In contrast, B cells stimulated with interleukin 4 display Fc epsilon R with high densities. Interestingly, IgE enhanced the Fc epsilon R expression induced by interleukin 4, suggesting that both interleukin 4 and IgE may be responsible for an increase in the expression of Fc epsilon R on B cells of N. brasiliensis infected mice.     

Characterization of human T cell-derived IgE-potentiating factor

We have previously shown that Fc epsilon receptor-positive (Fc epsilon R+) T cell lines from patients with the hyper IgE syndrome secrete IgE-binding factors which selectively enhance IgE but not IgG synthesis in cultures of B cells obtained from patients with allergic rhinitis but not from nonatopic subject. In the present study we have tested the effect of supernatants from Fc epsilon R+ T cell lines on a large panel of B cells from atopic patients (n = 20). We found that IgE synthesis was selectively enhanced only in B cell cultures in which there was ongoing spontaneous synthesis of IgE. The target of IgE-potentiating factor(s) was a large low-density B cell present in the circulation of responding atopic donors. In addition, we further characterized IgE-potentiating factors derived from Fc epsilon R+ T cell lines. The factor(s) fractionated into 2 peaks on Sephadex G-75 with approximate molecular masses of 15,000 and 60,000 kDa, and had affinity for lentil lectin but not for peanut agglutinin. Release of IgE-potentiating factor(s) was enhanced by the addition of exogenous human IgE to Fc epsilon R+ T cell cultures and was inhibited by tunicamycin, an inhibitor of N-glycosylation. These studies suggest a close homology between the physicochemical characteristics of human and rodent IgE-potentiating factors and the immune signals which modulate production of these IgE regulatory factors.     

Regulation of in vivo expression of Fc receptors for IgE (Fc epsilon R) on murine lymphocytes. II. Induction of Fc epsilon R and its inhibition in mice immunized with antigen

The induction of Fc receptors for IgE (Fc epsilon R) and its regulation were studied in BALB/c, SJL/J, and nude mice by a flow cytometric assay with the use of homologous monomeric IgE. Immunization of BALB/c mice with alum-absorbed antigen induced a remarkable increase in the expression of Fc epsilon R on spleen cells, whereas no enhancement of the Fc epsilon R expression was observed in SJL/J and nude mice after immunization. This increase was correlated with the elevation of serum IgE levels. However, the IgG antibody response, which is inducible even in SJL/J mice, was not associated with the induction of Fc epsilon R. The enhanced expression of Fc epsilon R in BALB/c mice observed in the primary or secondary IgE antibody response was detected in B cells with B220, surface IgM, and IgD, but not in T cells. The induction of Fc epsilon R in immunized BALB/c mice was inhibited by suppressive factor of allergy isolated from ascites fluids of SJL/J mice inoculated with complete Freund's adjuvant. In addition, both cyclophosphamide and prednisolone had an inhibitory effect on the induction of Fc epsilon R. These results suggest that the Fc epsilon R induction is inhibited not only by suppressive factor of allergy, which is effective in inhibiting the IgE antibody response selectively, but also by some immunosuppressive agents which are capable of suppressing all isotypes.     

IgE receptors on human lymphocytes. II. Detection of cells bearing IgE receptors in unstimulated mononuclear cells by means of a monoclonal antibody

A monoclonal antibody (mAb) specific for lymphocyte IgE receptors (ER) was employed in a rosette assay for the detection of cells bearing IgE receptors (Fc epsilon R). The specificity of the assay was documented by inhibition studies with soluble immunoglobulins (Ig) and anti-Ig antibodies. Moreover, similar results were obtained by employing the F(ab')2 fragment of mAbER instead of intact molecule. Circulating mononuclear cells isolated from normal or allergic adults and from umbilical cord blood contained approximately 8% of Fc epsilon R-bearing cells with values ranging from 0.3 to 17%. Tonsillar lymphocytes contained about 30% of Fc epsilon R+ cells. After the removal of adherent cells, there was a small but significant reduction of the proportion of Fc epsilon R+ cells. When mononuclear cells were separated into T and B cell fractions by two-cycle rosetting with 2-aminoethylisothiouronium bromide hydrobromide-treated sheep red blood cells, most of the Fc epsilon R+ cells were in the B cell fraction; however, a small proportion of Fc epsilon R+ was also found in the enriched T cells and double-labeling experiments confirmed that these cells were indeed T lymphocytes. Fc epsilon R+ cells were purified by rosetting with mAbER-coated erythrocytes and their phenotype was compared to that of Fc epsilon R- cells; Fc epsilon R+ cells contained about 90% of B cells (B1+) together with a small proportion of OKT3+, Leu 7+ and Mo2+ cells. The bulk of T cells, macrophages and natural killer (NK) cells was found in the Fc epsilon R- cells which contained fewer B cells than the fraction of Fc epsilon R+ cells. These data thus indicated that the great majority of Fc epsilon R-bearing cells are B cells but that a small proportion of NK cells, macrophages and T lymphocytes also express Fc epsilon R. Upon incubation at 37 degrees C, B cells lost their Fc epsilon R and this phenomenon was selectively inhibited by IgE; however, purified T cells seemed to express more Fc epsilon R after overnight incubation at 37 degrees C and this was not influenced by IgE. It is finally shown that the expression of Fc epsilon R is cyclic and that Fc epsilon R-bearing B cells do not represent a functionally distinct subpopulation of B lymphocytes.     

Fc-epsilon-receptor-bearing lymphocytes in patients with clonorchiasis

Expression of Fc-epsilon-receptors (Fc epsilon R) on peripheral blood lymphocytes was examined in patients with clonorchiasis. Mean serum IgE levels in these patients was 2,518 IU/ml. Fc epsilon R was detected by flow cytometry analysis with monoclonal antibody. The frequency of Fc epsilon R+ lymphocytes and the density of Fc epsilon R on the cells in the patients were similar to those in normal subjects. Most of the Fc epsilon R+ lymphocytes were B cells. The number of Fc epsilon R+ cells decreased after incubation in serum-free medium at 37 degrees C for 3 h. The amount of regulatory molecules of IgE production, IgE binding factor, was not significantly different between the patients and normal subjects. The number of Fc gamma R bearing T or B cells in the patients was also similar to that of normal subjects. These results indicate that the mechanisms of elevated serum IgE in patients with clonorchiasis might be different from other diseases with hyperglobulinemia E.     

Regulation of IgE receptor expression on human peripheral blood lymphocytes by lymphocytosis promoting factor (LPF), lectins and dexamethasone.

Using a monoclonal anti-human Fc epsilon R antibody (H107), we found that lymphocytosis promoting factor (LPF), phytohaemagglutinin (PHA-P) and Concanavalin A (Con A) could induce Fc epsilon R, detected by immunofluorescence study, on normal human peripheral blood lymphocytes without IgE. The number of Fc epsilon R bearing lymphocytes was increased by stimulation with 3, 10 and 10 micrograms/ml of LPF, PHA-P and Con A, respectively, from 6.0 +/- 3.0/1000 cells to 26.0 +/- 7.9, 54.0 +/- 6.7 and 24.8 +/- 7.1/1000 cells, respectively. Although the induction of Fc epsilon R occurred neither in the separated T-enriched fraction (TEF) nor the T-depleted fraction (TDF), it recovered when the two fractions were mixed. The cell free supernatants from TEF stimulated with LPF or PHA-P could increase Fc epsilon R(+) cells in TDF, whereas those from TDF failed to increase them in TEF. The results suggest that the induction of Fc epsilon R occurs mainly on B lymphocytes by the soluble factor(s) formed by T cells stimulated with LPF or PHA-P. The induction of Fc epsilon R by stimulants was completely inhibited by 10(-6) M dexamethasone. It was demonstrated that the effects of dexamethasone on lymphocytes were dual: one was on B cells to inhibit responsive increases of Fc epsilon R, and the other was on T cells to suppress the formation of the soluble factor(s) which induced Fc epsilon R on B cells.     

The expression of IgE Fc receptors on lymphocytes of allergic patients

Peripheral blood lymphocytes from nonatopic subjects and atopic patients were analyzed for cells expressing Fc receptors for IgE (Fc epsilon R). Nonatopic humans and atopic patients in remission had approximately 1 percent of Fc epsilon R+ peripheral blood lymphocytes. Usually greater than 99 percent of these cells were mIgM+/mIgD+ B cells. However, in approximately 10 percent of nonatopic and atopic subjects a transient increase of Fc epsilon R+ lymphocytes to 3-6 percent was observed in the absence of any disease manifestations and measurable changes in the serum IgE level. At times of increased numbers of peripheral blood Fc epsilon R+ lymphocytes, up to 1 percent Fc epsilon R+ positive cells were detected in isolated T cell preparations. The Fc epsilon R+ T cells reacted with the monoclonal antibody Lyt 3 to the sheep erythrocyte receptor of human T cells but not the anti-T cell antibody OKT3, and fractions also with the monoclonal antibodies OKT8 (cytotoxic and suppressor T cells) and OKM1, which binds to an antigen present on monocytes and a subpopulation of T cells and large granular lymphocytes. No OKT4+ (helper T cells) Fc epsilon R+ cells were detected. The reactivity with monoclonal antibodies to T cell subsets of the Fc epsilon R+ T cells paralleled the reactivity of the IgG Fc receptor positive T cells. In contrast to patients with allergic rhinitis and asthma, patients with severe atopic dermatitis or the Hyper IgE Syndrome always had significantly elevated percentage of Fc epsilon R+ lymphocytes (4-10 percent), which were almost entirely B cells since less than 0.1 percent Fc epsilon R+ T cells were detected in these patients. Atopic dermatitis patients receiving systemic corticosteroid treatment had only 0.2 percent Fc epsilon R+ lymphocytes which was significantly less than the 1 percent of the nonatopic control donors. Attempts to define the function of Fc epsilon R on human B and T lymphocytes have been unsuccessful thus far; however, the increase of Fc epsilon R+ cells associated with atopic disease in man and parasitic infections in rats and mice suggest that Fc epsilon R+ lymphocyte may be involved in the IgE isotype regulation.     

Presence of antigenic determinants common to Fc IgE receptors on human macrophages, T and B lymphocytes and IgE-binding factors

The present study indicates that two Mab specific to Fc epsilon R (MabER) on human B lymphocytes also react with Fc epsilon R on macrophage and T-cell lines. More importantly, it is also shown that MabER cross-react with IgE-BFs derived from B, T and macrophage cell lines. This conclusion is supported by the following observations: the binding of MabER to Fc epsilon R-bearing cells is blocked by the CSN of T, B and macrophage Fc epsilon R-bearing cell lines, known to contain IgE-BFs as shown by their inhibition of rosette formation between IgE-coated erythrocytes and Fc epsilon R-bearing cells; the material purified from the CSN of each Fc epsilon R(+) cell lines by affinity chromatography on MabER-Affi-gel blocks the rosetting of U937 cells with IgE but not with IgG-coated erythrocytes; the same affinity-purified material inhibits the binding of 125I-IgE to a selected anti-IgE Mab (Mab 75); and the CSN of Fc epsilon R(+) cells but not of Fc epsilon R(-) cells reacts in a solid-phase sandwich radioimmunoassay with two MabER (135-176), and their reactivity is significantly retained on IgE-Affi-gel from which it may be recovered by glycine elution. This RIA is not influenced by any class of Ig, including IgE, employed at a final concentration of 100 micrograms/ml. Human serum also reacts in the RIA, and parallel dilution curves are obtained with different CSN and human sera. The RIA proved to be a reproducible and sensitive method to quantify human IgE-BFs. The expression of the same antigenic determinants on Fc epsilon R from T, B and macrophages as well as on the IgE-BFs secreted by these cells indicates structural homology between IgE receptors and IgE-FBs and suggests that they are encoded by the same gene.     

Induction and function of Fc epsilon RII on YT cells; possible role of ADF/thioredoxin in Fc epsilon RII expression.

The regulation of low-affinity Fc receptor for IgE (Fc epsilon RII) and the characteristics of both membrane and soluble forms of Fc epsilon RII were studied using YT cell line. We found that YT cells, a human NK like cell line, expressed Fc epsilon RII after IL-1 stimulation. Cross-linking of Fc epsilon RII on IL-1-stimulated YT cells as well as the transfectant of Fc epsilon RII-cDNA (YTSER) resulted in the up-regulation of IL-2R alpha (p55/Tac). A 59 kDa protein phosphorylated at tyrosine residues was co-immunoprecipitated with Fc epsilon RII from YTSER lysate using H107 anti-Fc epsilon RII mAb. YTSER not only expressed Fc epsilon RII on their surface but also secreted soluble form of Fc epsilon RII (sFc epsilon RII/sCD23; IgE binding factor). Affinity purification revealed that sFc epsilon RII released from YTSER is heterogeneous and consisted of several proteins differing in molecular weight. Both EBV+ B cells and HTLV-1+ T cells are high producers of ATL derived factor (ADF)/thioredoxin (TRX) and express Fc epsilon RII and IL-2R alpha respectively. To clarify the mechanism of Fc epsilon RII and IL-2R alpha induction by ADF/TRX, we examined the effect of ADF/TRX on the bindability of nuclear factor kappa B (NF-kappa B), which is known to regulate IL-2R alpha gene expression. In the gel shift assay, ADF/TRX was shown to enhance the bindability of NF-kappa B to its responsive element.     

IL-4 production by T depleted cells from Nippostrongylus brasiliensis infected mice

Interleukin-4 (IL-4) is known to be involved in both the in vivo IgE response and the elevated B cell IgE Fc receptor (Fc epsilon R11) expression seen after a parasite infection. To further analyze the relationship between Fc epsilon R11 expression and IL-4 production, purified B cells from uninfected, Nippostrongylus brasiliensis (Nbr) infected and from goat anti-mouse IgD (GaM delta) injected mice were isolated on various days post-treatment. The Fc episolon R11 levels on purified B cells from normal mice decreased after an overnight culture in media alone and addition of IL-4 to these cultures resulted in a 4 to 13-fold enhancement of Fc epsilon R11 levels. In contrast, the Fc epsilon R11 levels on B cells from Nbr infected mice were elevated after an overnight culture in media alone and addition of IL-4 did not further enhance the already upregulated Fc epsilon R11 levels. Overnight culture of purified B cell blasts from Nbr infected mice in the presence of an anti-IL-4 monoclonal antibody (11B11) caused the elevated Fc epsilon R11 levels to return to levels seen in normal mice, without affecting the Fc epsilon R11 levels on purified Go or B cell blasts from uninfected mice or Go B cells from Nbr infected mice. 11B11 also inhibited the elevated Fc epsilon R11 levels on highly purified B cells obtained by FACS sorting the non-adherent spleen cell population for class II+ cells. In contrast to Nbr infection, the Fc epsilon R11 levels on B cells were downregulated in the GaM delta injected mice. However, analogous to the Nbr system, the Fc epsilon R11 levels were unresponsive to the addition of exogenous IL-4. This study indicates that IL-4 production is seen in T depleted splenocytes and that this alternate source of IL-4 serves to maintain the elevated Fc epsilon R11 levels on B cells.     

The IgE system

The data presented herein indicate that IgE synthesis results from a complex interaction between T cells, B cells, and monocytes, under the control of T cell-derived and monocyte-derived lymphokines (IL-4, IL-5, and IL-6). In addition to their direct effects on IgE synthesis, these lymphokines have other functions, many of which are related to IgE. IL-4 is not only a crucial signal for the induction of IgE synthesis, but also a growth factor for murine mast cells together with IL-3, a product of the same T cell clones. Furthermore, IL-4 is a potent inducer of Fc epsilon R2b expression on both B cells and monocytes. On the other hand, IL-4-secreting T cells also produce IL-5, which promotes B cell maturation and induces differentiation of eosinophil precursors. Eosinophils, in turn, express CD23 when activated; thus, the activation of the IgE system leads to increased IgE synthesis and to enhanced expression of Fc epsilon R1 (on mast cells) and Fc epsilon R2 (on B cells, monocytes, and eosinophils). This in turn results in increased receptor-ligand interactions, with release of different chemical mediators involved in the pathogenesis of allergy. Such considerations suggest that the regulation of IgE synthesis should be viewed in the wider context of the IgE system, a multi-component network in which different cell types are functionally integrated mainly through lymphokine-mediated signals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Secretion of IgE-specific potentiating factors by human Fc epsilon R+ T cell lines

In the present study, Fc epsilon R+ and Fc epsilon R- T cells were isolated from patients with the hyper-IgE syndrome and maintained in long-term cultures with interleukin 2. Supernatants from the Fc epsilon R+ but not from the Fc epsilon R- T cell lines enhanced IgE but not IgG synthesis in B cells derived from patients with allergic rhinitis. There was, however, no induction of IgE synthesis in B cells from nonatopic donors. The IgE-potentiating factors bound to IgE-Sepharose but not to IgG-Sepharose. The target B cells for these IgE binding factors appear to be preactivated IgE-bearing B cells.     

IgE Fc receptor positive T, B and NK cells in patients with the hyper-IgE syndrome

Patients with the hyper-IgE syndrome have greatly elevated percentages of IgE Fc receptor (Fc epsilon R)-positive B cells, but they have less than 0.1% Fc epsilon R+ T cells (T epsilon cells) and few, if any, Fc epsilon R+ natural killer cells. They also have markedly decreased numbers of IgG receptor positive (Fc gamma R+) T cells (T gamma cells). Patients with the hyper-IgE syndrome resemble in this respect patients with severe atopic dermatitis. Since a portion of T epsilon and T gamma cells of mildly atopic patients react with monoclonal antibody OKT8, they may have a suppressor function. However, whether the low number of T epsilon cells is responsible for the high IgE serum level in hyper-IgE syndrome and atopic dermatitis patients remains to be demonstrated. Attempts to obtain a reliable assay for human IgE synthesis in vitro to investigate the function of Fc epsilon R-positive lymphocytes proved to be difficult. Even isolated B cells from atopic donors seldom produced more than twice the quantity of IgE released from cells incubated in the presence of the protein synthesis inhibitor cycloheximide.     

The expression of B cell surface receptors. III. The murine low-affinity IgE Fc receptor is not expressed on Ly 1 or 'Ly 1-like' B cells.

The distribution of IgE FcR (Fc epsilon R)-positive and -negative B cells was examined in normal adult mice. Using three-color flow cytometry, the expression of the Fc epsilon R was analyzed on various B-cell subsets present in the peritoneum and spleen. The results demonstrate that in the peritoneal cavity, the Fc epsilon R is not expressed on the large majority of Ly 1+ B cells and Ly 1-, Mac 1+ sister B cells. The receptor is present, however, on the small number of conventional B cells residing in the peritoneum. Although interleukin 4 (IL-4) can increase the levels of the Fc epsilon R on conventional B cells, incubation of Ly 1 and sister B cells with IL-4 did not result in the expression of the Fc epsilon R. When examining B cells present in the spleen, a small subset of B cells was consistently found to be Fc epsilon R-. These Fc epsilon R- cells were IgM-bright, IgD-dull and largely Ly 1- and Mac 1-negative. Staining of splenic tissue sections revealed that the Fc epsilon R- B cells were primarily localized to the marginal zones, whereas the Fc epsilon R+ B cells were found in the follicles. Taken together, the results indicate that the Fc epsilon R may be a useful marker in delineating the various B-cell subsets. In the peritoneum, the Fc epsilon R appears to discriminate conventional B cells from those of the Ly 1/sister lineage, and in the spleen it is likely to distinguish resting follicular B cells from Ly 1/sister and marginal zone B cells.     

IgE Fc receptor positive T and B lymphocytes in patients with the hyper IgE syndrome.

The percentages of peripheral blood lymphocytes (PBL), bearing Fc receptors for IgE (Fc epsilon R) and IgG (Fc gamma R) were determined in four patients with the hyper IgE syndrome by a rosette assay employing IgE and IgG coated fixed ox erythrocytes. The patients had 8 +/- 3% Fc epsilon R+ and 13 +/- 8% Fc gamma R+ PBL, compared to 1.2 +/- 1% Fc epsilon R+ and 17 +/- 4% Fc gamma R+ PBL for control donors. T cells were isolated by rosetting with neuraminidase treated sheep erythrocytes (EN). Indirect immunofluorescence with Lyt 3 monoclonal antibody (MoAb) to the sheep erythrocyte receptor, followed by rosetting for Fc epsilon R and Fc gamma R showed that the patients' T cells contained less than 0.1% Fc epsilon R+ and 1.4 +/- 0.2% Fc gamma R+ cells; T cells from the control subjects contained less than 0.1% Fc epsilon R+ and 11 +/- 4% Fc gamma R+ cells. The non-T (EN rosette depleted) cells of the patients included 56 +/- 18% sIgM+/sIgD+, 45 +/- 9% Fc epsilon R+ and 35 +/- 27% Fc gamma R+ cells. Indirect immunofluorescence with MoAb to IgM, IgD, and NK cells (antibody B73.1) followed by rosetting for Fc epsilon R and Fc gamma R, indicated that 92 +/- 2% of the Fc epsilon R+ cells and 9 +/- 7% of the Fc gamma R+ cells were B cells (mu+/delta+), while 3 +/- 4% of the Fc epsilon R+ and 30 +/- 23% of the Fc gamma R+ cells were NK cells (B73.1+). Thus, most of the Fc epsilon R+ non-T cells were B cells, and only a small fraction appeared to be NK cells. On the other hand, Fc gamma R+ B cells were outnumbered by Fc gamma R+ NK cells (B73.1+) by three to one. The data indicate that patients with the hyper IgE syndrome have increased numbers of Fc gamma R+ PBL, most of them being B cells, whereas their T cells contain less than 0.1% Fc epsilon R+ cells.     

Human Fc epsilon R II and IgE-binding factors

The low-affinity receptor for IgE (Fc epsilon R II) is mainly expressed on B lymphocytes, although it may also be found on some monocytes, eosinophils, and platelets. The presence of Fc epsilon R II on T cells is still controversial, but our results demonstrate that it is expressed on some HTLV-I-transformed T lymphocytes, and they strongly suggest that it may be found on a small proportion of normal T cells. Fc epsilon R II is a 45-KD glycoprotein containing one N-linked carbohydrate of complex type, O-linked carbohydrates, and sialic acid residues. Fc II is cleaved into soluble fragments with molecular weights of 37, 33, 25, and 12 KD, the first three retain the ability of binding to IgE, i.e., they are IgE-binding factors (IgE-BFs). The enzymes involved in their proteolytic cleavage are cell bound. The cDNA coding for Fc epsilon R II was cloned and functionally expressed. The predicted sequence has no homology with that of murine IgE-BFs which are of T cell origin. However, there is a striking homology with several animal lectins, and since the IgE-binding site is located in the homology region, it is possible that it binds to IgE via the carbohydrates expressed on the Fc region of this immunoglobulin. The expression of Fc epsilon R II on B cells and the release of IgE-BFs are upregulated by interleukin 4 and suppressed by gamma and alpha interferons.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of the low-affinity receptor for IgE on murine B cells by using membrane impermeant cross-linking reagents

The murine B lymphocyte receptor for the Fc portion of IgE (Fc epsilon R) was further characterized by using the membrane impermeable cross-linking reagents 3,3'-dithiobis-(sulfosuccinimidyl) proprionate (DTSSP) and bis-(sulfosuccinimidyl) suberate (BS3). IgE could be cross-linked to the Fc epsilon R on the intact cells with either reagent and, in addition, up to 10% of the B cell surface immunoglobulin (sIg; both IgM and IgD) was also found to cross-link to the IgE/Fc epsilon R complex. Analysis of isolated sIg/IgE/Fc epsilon R complexes indicated that about 60% of the Fc epsilon molecules were becoming cross-linked to sIg. Thus, the data suggest that on the intact murine B cell the Fc epsilon R is frequently in close association with sIg. The murine B cell Fc epsilon R was also examined for the presence of receptor-associated proteins that are buried in the membrane. Advantage was taken of the membrane-impermeant nature of DTSSP and BS3. IgE was cross-linked to the Fc epsilon R on intact cells by using the disulfide-cleavable DTSSP and following solubilization with nonionic detergent; BS3 was used to cross-link possible internal membrane components to the Fc epsilon R. In these experiments, the high-affinity Fc epsilon R on rat basophilic leukemia (RBL) cells could be cross-linked to a nonreducible high molecular weight complex of 100 kilodaltons. However, when intact murine B cells were treated with DTSSP, solubilized and treated with BS3 in the same manner as indicated above, no evidence was found for the presence of membrane-buried receptor-associated proteins with the B cell Fc epsilon R. Thus, these data further support the concept that there may be little relationship between the high-affinity mast cell/basophil Fc epsilon R and the low-affinity lymphocyte Fc epsilon R.     

Expression of Fc epsilon receptors on activated human T lymphocytes

Our results clearly demonstrate that the low-affinity receptor for IgE (Fc epsilon R) is an activation antigen transiently expressed on a subpopulation of human T lymphocytes. It can be selectively induced by stimulation with certain antigens or lectins, but it is not found on resting T cells. The increased numbers of activated Fc epsilon R+ T cells observed after stimulation of peripheral blood mononuclear cells (PBMC) from bee venom allergic patients with the specific allergen phospholipase A2 (PLA2) suggest that Fc epsilon R+ T cells might very well be involved in the regulation of the human IgE response against the respective antigen. These results were obtained by the use of two monoclonal antibodies, M-L25 and M-L47, which were raised against the human low-affinity Fc epsilon R in our laboratory. After stimulation of PBMC with phytohemagglutinin a peak of 7.6 +/- 6% Fc epsilon R+ T cells was observed on day 3, with pokeweed mitogen of 0.8 +/- 0.8% on days 2 and 3, and with concanavalin A of 0.6 +/- 0.7% Fc epsilon R+ T cells on day 2. Stimulation of PBMC with tetanus toxoid (TT) induced Fc epsilon R on maximally 0.6 +/- 0.8% of the total T cells (day 4), stimulation with purified protein derivative from tuberculin (PPD) on 0.2 +/- 0.6% of the T cells (day 2). In contrast to these antigens, stimulation of PBMC from bee venom allergic patients with PLA2 induced as a peak 2.5 +/- 2.5% of the total T cells to express Fc epsilon R (day 5), although the stimulated T cell population was much smaller than with TT or PPD, as was shown by their stimulation indices. The allergen-stimulated Fc epsilon R+ T cells were exclusively T4+. The Fc epsilon R-expression index was determined, which for a specific antigen or lectin correlates the percentage of Fc epsilon R+ T cells to the stimulated T cell population, respectively.     

Soluble Fc epsilon R II levels in normal children and patients with immunodeficiency diseases

CD23 is expressed on mature B cells and is identical to a low-affinity IgE Fc epsilon receptor type II (Fc epsilon R II). The C terminal portion of CD23 is released to the serum as soluble Fc epsilon R II (sFc epsilon R II), which may be involved in regulation of IgE synthesis. We studied sFc epsilon R II levels in normal children and in patients with immunodeficiencies, including common variable immunodeficiency (CVI), partial DiGeorge syndrome, and immunodeficiency associated with ectodermal dysplasia to examine the relationship of sFc epsilon R II levels to B cell numbers and other immunoparameters. Serum Fc epsilon R II levels are higher in younger children (younger than 3 years) and decline gradually with age. In 11 patients with CVI with normal numbers of B cells (greater than 6%), sFc epsilon R II levels were comparable to that of control subjects. Five patients with CVI with deficiencies of peripheral B cells had levels of sFc epsilon R II similar to levels of control subjects. In all but one patient with partial DiGeorge syndrome, sFc epsilon R II levels were not significantly elevated, despite the presence of elevated peripheral B cell numbers. Of six patients with ectodermal dysplasia, four demonstrated increased Fc epsilon R II levels, a finding not correlated with serum IgE levels or with peripheral eosinophil or B cell numbers.