The presence of IgE on the surface of lymphocytes in nasal polyps

The presence of IgE-bearing lymphocytes in nasal polyps was correlated with the patients' atopic status. Following surgical removal, the polyp tissue was treated with hyaluronidase and a single-cell suspension was obtained. Lymphocytes were isolated by gradient centrifugation, and the fluorescent antibody technique was used to study the presence of various immunoglobulin markers on the surface of lymphocytes. The presence of IgE-bearing cells was correlated with serum IgE levels, history of allergy, and skin test reactions. Patients with a positive atopic history had intermediate to high serum IgE levels. There was no correlation between IgE level and skin reactivity in these patients. Good correlation was obtained between the number of IgE-bearing cells in nasal polyps and serum IgE levels in atopic patients. The IgE-bearing cells represented 10 to 40 per cent of total immunoglobulin-bearing lymphocytes. No IgE-bearing cells were detected in five of six patients with a negative atopicf history and negative skin tests. Thus IgE may be synthesized within nasal polyps of atopic patients, and the polyps in atopic patients may have a different etiology from those in nonatopic patients.     

A high-affinity monoclonal anti-IgE antibody for depletion of IgE and IgE-bearing cells

Background:  We have identified a monoclonal anti-human immunoglobulin E (IgE) antibody, which recognizes FcepsilonRI-bound IgE and prevents binding of IgE to FcepsilonRI. In this study, we assessed the binding kinetics and affinity of monoclonal antibody 12 (mAb12) for IgE and investigated whether mAb12 can be used for depletion of IgE and isolation of IgE-bearing cells from peripheral blood.
Methods:  Binding kinetics and affinity for IgE were studied using Biacore surface plasmon resonance technique experiments. IgE antibodies were depleted from serum using sepharose-coupled mAb12 and IgE-bearing cells were enriched from heparinized blood samples with mAb12. The extent and biological relevance of IgE depletion were studied by quantitative IgE measurements and basophil histamine release experiments. Specific binding of mAb12 to IgE-bearing cells (basophils, mast cells, IgE-secreting plasma cells) was demonstrated by FACS.
Results:  Monoclonal antibody 12 shows rapid association (k_a = 5.46e5/Ms) with IgE, almost no dissociation (k_d = 8.8e−5/s) and an affinity for IgE (K_D = 1.61e−10 M), which is as high as that of FcepsilonRI. Immobilized mAb12 could be used to deplete IgE antibodies and isolate IgE-bearing cells from peripheral blood in a single-step procedure.
Conclusions:  Monoclonal antibody 12 is a high affinity anti-human IgE antibody, which efficiently removes IgE and IgE-bearing cells from peripheral blood and may thus be used for extracorporeal depletion of IgE and IgE-bearing cells.     

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.     

Differentiation of membrane IgE+ rat B cells into IgE-secreting cells.

Rat spleen cells were stimulated with pokeweed mitogen (PWM) and the IgM and IgE responses were assessed. An enrichment of the cell suspension with IgE-bearing cells before stimulation resulted in an increase in the number of IgE-secreting cells. A decrease of the number of IgE-secreting cells was found after depletion of IgE- or IgM-bearing cells, but not those bearing IgD molecules on their membranes, before stimulation. Moreover, the stimulation of membrane IgE on B cells with anti-IgE antibodies was shown to increase the number of IgE-secreting cells after PWM-induced differentiation in vitro. In vivo, it was also observed that a single injection of anti-IgE antibodies can induce the differentiation of IgE-secreting cells. These results demonstrate the presence of IgE(+)-IgM (+)-IgD- B cells in the rat that are responsive to PWM-induced differentiation into IgE-secreting cells. They indicate a pre-commitment of these cells at a stage where they still express IgM on their surface. IgE molecules on the cell membranes play a role in their differentiation.     

The Langerhans cell: an underestimated cell in atopic disease

Langerhans cells (LC) are very potent antigen-presenting cells. In atopic disorders such as allergic rhinitis and atopic dermatitis LC are known to bear IgE surface molecules. IgE-positive LC can bind allergen and present it to T lymphocytes to induce an allergen-specific T-cell response and IgE synthesis. Therefore, IgE-bearing LC might play an important role in the triggering of the immune system to maintain ongoing IgE synthesis. The importance of the IgE-bearing LC in atopy has not been assessed but deserves further investigation to find out more about the part played by these cells, not only in the atopic disorders described here but also in others such as gastrointestinal allergy and allergic asthma.     

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-positive duodenal mast cells in patients with food-related diarrhea

Thirty-seven consecutive adult patients with a history of adverse reactions to foods, manifested mainly as diarrhea, were investigated with skin prick test (SPT), IgE levels in serum, and the presence of IgE bound to mast cells in duodenal biopsy specimens. Nineteen percent had increased serum IgE levels indicating the presence of atopic disease and in 35% positive SPTs for one or several allergens related to their gastrointestinal symptoms were found. In 92% of the patients with positive SPTs to food, IgE-positive mast cells in duodenal biopsy specimens were found, twice as many as in patients with negative SPTs (47%). Forty-two percent of normal individuals without any adverse reactions to foods had IgE-bearing mast cells in their intestinal mucosa. The results showed that 'arming' of mast cells with IgE locally was relatively prominent but not consistent in the gut of patients with food-related diarrhea suggested to be allergic by positive SPT. Intestinal IgE-mediated hypersensitivity reactions could be of pathogenetic significance in most of these patients. However, since also half of the normal individuals were found to have IgE-positive intestinal mast cells, this phenomenon presumably also reflects a normal physiological defence mechanism.     

Isotype-specific human B lymphocytes that produce immunoglobulin E in vitro when stimulated by pokeweed mitogen

Normal human peripheral blood lymphocytes can be stimulated by PWM and T cells to produce IgE in vitro. Between 4% and 10% of circulating B lymphocytes were found to bear noncytophilic membrane IgE. In contrast, limiting dilution analysis of reactive B cell precursor frequency revealed that only about 1 in 50,000 B cells was responsible for the synthesis of IgE in vitro. Variation in the precursor frequency was primarily responsible for differences in the amount of IgE produced in vitro by different individuals while the amount of IgE produced per precursor cell was relatively constant. B lymphocytes with surface IgM, IgG, or IgE were depleted. Depletion of IgE-bearing cells decreased IgE production 70% to 83% while IgM-bearing-cell removal lowered IgE synthesis by 59% to 72%. Dual removal of IgM- and IgE-bearing cells did not give an additive effect. IgE- and IgM-positive B cells showed enhanced IgE synthesis of 327% and 182%, respectively. The consequences of crosslinking different surface isotypes on subsequent IgE production were also assessed. Anti-? treatment inhibited IgE production up to 68% while anti-μ, treatment decreased it by up to 38%. Thus only a very limited subset of B lymphocytes is responsible for PWM and T cell-dependent IgE synthesis in vitro and these cells probably bear both surface IgE and IgM.     

银屑病发病机制中IL-17关键轴与IgE的相互作用

银屑病是一种免疫介导的多基因遗传性皮肤病,其发病机制尚未明确,目前认为可能是遗传、环境和免疫学因素共同作用的结果,在免疫机制方面,白细胞介素(interleukin,IL)-23 /辅助T细胞(helper T cells,Th)17途径已被确定为关键轴,促炎细胞因子IL-17A是其主要作用因子。有研究表明,银屑病患者血清中免疫球蛋白(immune globulin,Ig)E浓度增加。与非病变皮肤相比,病变皮肤有更多的特异性IgE的Fc段高亲和力受体I (receptor I for the Fc region of IgE,FceRI)相关细胞,包括肥大细胞,表皮树突状细胞,朗格汉斯细胞和巨噬细胞,现就银屑病发病机制中IL-17关键轴与IgE的相互作用进行综述。     

Local allergic reaction in food-hypersensitive adults despite a lack of systemic food-specific IgE

BACKGROUND: Objective tools are lacking for the diagnosis of local gastrointestinal inflammatory reactions in skin prick test (SPT)-negative and serum IgE antibody (s-IgE Ab)-negative patients with suspected food allergy. OBJECTIVE: The purpose of this investigation was to evaluate the presence of eosinophils, T cells, local IgE-bearing cells, IL-4, and IFN-gamma in small intestinal biopsy specimens from adult SPT-negative/s-IgE Ab-negative patients with food allergy during symptomatic and nonsymptomatic periods. METHODS: Fourteen patients with food allergy-related gastrointestinal symptoms confirmed by double-blinded, placebo-controlled food challenge (DBPCFC) were investigated. Eleven of the patients were SPT-negative and s-IgE Ab-negative. Sex- and age-matched healthy volunteers were used as controls. Duodenal biopsies were studied with immunostaining through use of a panel of mouse monoclonal antibodies specific for eosinophils, CD3, CD4, CD8, IgE, IL-4, and IFN-gamma. RESULTS: Significant increases in numbers of MBP+ eosinophils, IgE-bearing cells, and T cells were found in the duodenal mucosa of the patients when they were symptomatic in comparison with when they were asymptomatic and in comparison with healthy controls. Numbers of IL-4+ cells were increased and numbers of IFN-gamma+ cells were reduced in the patients when they were symptomatic in comparison with when they were asymptomatic and in comparison with the controls. There were no differences in total s-IgE levels between any of the groups. CONCLUSION: A significant correlation was found between the appearance of symptoms of food hypersensitivity and the duodenal presence of IgE-bearing cells, activated eosinophils, and T cells in patients with negative SPT results and negative s-IgE Ab to the offending food. We suggest that a localized IgE-mediated response caused the gastrointestinal symptoms seen in these patients.     

Immunopathology of skin lesions in Kawasaki disease]

Histopathological findings of skin lesions in Kawasaki disease (KD) have been characterized by extensive edema associated with the dilatation of small vessels in the papillary dermis. Although the inflammation in KD skin lesions was exudative in nature, neutrophil emigration was slight and most of the infiltrates were mononuclear cells. Immunofluorescent studies using skin biopsy specimens from 10 patients with KD aged from six months to eight years and seven months showed that the infiltrating cells in the dermis and epidermis were mainly composed of CD 3+ T cells and Leu M3+ macrophages, but not B cells. In double immunofluorescence staining with combinations of anti HLA-DR, CD4 and CD8 monoclonal antibodies, the infiltrating T cells were mainly CD4+ HLA-DR+ T cells. Leu 23% cell were also positive on these cells, thereby suggesting those to be activated. Studies of skin specimens obtained from the site of BCG vaccinations in patients with KD showed basically similar but more extensive lesions. As a control, the infiltrating cells in the dermis from patients with measles were examined. In contrast to KD, these cells were mainly CD8+ T cells. Together with the findings that the keratinocytes in the epidermis were positive for HLA-DR, the skin lesions in KD appear to be similar to those found in delayed type hypersensitivity. Thus, macrophages and helper T cells may play a crucial role in the pathogenesis of KD.     

Immunoglobulin E synthesis in parasite infection.

The infection of rats with Nippostrongylus brasiliensis (Nb) larvae enhanced IgE synthesis prior to the formation of IgE antibody specific for parasite antigens. As early as the eighth to tenth day of infection, IgE-bearing lymphocytes appeared in lymph nodes and spleen. At this time, even bone marrow contained IgE-bearing blast cells. The IgE-forming plasma cells were detected in the lymph nodes and spleen at the tenth day and their number reached maximum at the fourteenth day. The proliferation of IgE-bearing lymphocytes in the lymphoid tissues was observed in neonatally thymectomized animals, indicating that T cells are not essential for the development of IgE-B cells in the infected animals. It appears, however, that T cells are involved in the differentiation of IgE-B cells to IgE-forming plasma cells. Nonspecific proliferation of IgE-B cells and their differentiation to IgE-forming plasma cells may explain potentiation of IgE antibody formation against unrelated antigens after infection with N. brasiliensis (Nb). It was also found that T cells specific for parasite antigens were primed by infection with Nb. Evidence was obtained tht Nb-specific T cells raised by the infection collaborated with hapten-specific IgE-B cells and IgG-B cells for the secondary antihapten antibody responses. By contrast, T cells primed by immunization with parasite antigen included in alum exerted the helper function for IgG antibody response but failed to collaborate with IgE-B cells. Dissociation between the helper function for IgE and IgG antibody response indicated that parasite infection generated a favorable condition for priming T cells for the IgE antibody response.     

Immunopathology of atopic dermatitis

Conclusions In summary, AD is a chronic inflammatory skin disease associated with increased IgE production, eosinophilia, increased mast cell number and increased expression of the CD23 low-affinity IgE receptor on mononuclear cells. The basis of these immunologic defects appear to be the result of increased numbers of IL-4-, IL-5-producing T_h2 cells. This results in an overstimulation of their IL-4-IL-4 receptor pathway that is accompanied by a relative deficiency of IFN- production.The acute manifestations of AD occur as the result of mast cell degranulation that can be triggered by a variety of stimuli including allergens, bacterial toxins or histamine-releasing factors secreted by activated T cells and macrophages. Mast cells release not only histamine which contributes to the acute sensation of pruritus but also cytokines and chemotactic factors which induce leukocyte adhesion molecules and attract inflammatory cells into the local tissue sites [3, 8, 26, 34].The chronic manifestations of AD are probably the result of sustained cellular activation. The cellular infiltrate in the chronic lesion of AD consists primarily of T cells and macrophages [29]. T lymphocytes play two important roles in allergic responses: First, as already discussed they play a critical role in the regulation of IgE responses. Second, there is increasing evidence that they may play an important role in the control of the inflammatory response. This is orchestrated through organization of the cellular composition of the inflammatory cell infiltrate by the release of various cytokines. The release of IL-3, IL-5, and GM-CSF have pronounced effects on the differentiation and activation of eosinophils [52]. IL-3 promotes the growth of mucosal mast cells and stimulates basophil and mast cell histamine release. IL-4 promotes the growth of murine mast cells [18] and induces the expression of CD23 on macrophages [71]. The release of cytokines may, thus, account for the observation of increased mast cell number in chronic eczematoid lesions.Taken together, these observations suggest that the AD skin lesion is characterized by the infiltration of inflammatory cells which differ from those infiltrating into a type IV contact-sensitivity reaction. The sustained immune activation, which results in chronic AD, may be the result of an underlying T cell defect, e.g., decreased IFN- production following the activation of lymphocytes with a variety of stimuli. Treatment of patients withh rIFN- down-regulates the IL-4-IL-4 receptor pathway which is overstimulated in this disease. It is important to note, however, that although many immunopathologic features of AD have recently been elucidated, the fundamental etiology of these abnormalities remains to be unraveled.     

In vivo arming of cutaneous mast cell receptors by IgE released from macrophages

Intracutaneous injection of purified peritoneal macrophages harvested from ovalbumin (OVA)-hypersensitive high-IgE-responder BN rats into naive animals sensitised the injection sites for subsequent OVA-specific passive cutaneous anaphylaxis (PCA) reactions. The underlying mechanism(s) were investigated using a macrophage cell line (WEHI 265.1), which exhibited comparable sensitising activity in rat or mouse skin, after initial pulsing in vitro with antiserum rich in OVA-specific IgE. Transfer of OVA-hypersensitivity by the cell line (1) was IgE-dependent and did not occur when the cells were pre-exposed to antiserum containing OVA-specific IgG alone, (2) was blockable by saturation of cell surface receptors in the recipient with myeloma IgE (but not myeloma IgG), and (3) did not occur in mast cell-deficient mice carrying the W/Wv mutation, in contrast to their normal heterozygous littermates which developed marked OVA-hypersensitivity at the injection site. These results are consistent with arming of IgE-receptors on cutaneous mast cells by IgE antibody released from macrophages, and hint at a possible role for phagocytes in amplifying IgE-mediated reactions in tissues.     

Bacterial Histamine Release by Immunological and Non-Immunological Lectin-Mediated Reactions

The mechanisms of bacteria-induced histamine release were examined in vitro in human leukocytes and rat mast cells. Three types of bacterial responders were found. In persons with IgE-bearing basophilocytes bacterial histamine release could be triggered by two different mechanisms, an IgE-dependent mechanism where removal of IgE abolished the release and a non-immunological mechanism where this was not the case. In responders with no IgE-bearing cells bacterial histamine release was caused by a non-immunological mechanism. The non-immunological mechanism was further substantiated by release in isolated mast cells from germ-free rats. These experiments suggest a direct interaction between bacteria and target cell, and experiments with multi-washed bacteria and bacteria cell wall preparations indicate the possibility of the bacteria wall interacting with the target cell. It is probable that the non-immunological mechanism depends on lectin-mediated reactions, since bacteria-induced histamine release was inhibited by lectin-binding sugars as is release caused by plant lectins.     

Asiasari Radix Inhibits Immunoglobulin E Production on Experimental Models In Vitro and In Vivo

Immunoglobulin (Ig) E is the principal Ig involved in immediate hypersensitivities and chronic allergic diseases. The hallmark of these disorders is increased IgE production. The effect of an aqueous extract of the roots of Asiasari radix (ARAE) on an in vivo and in vitro IgE production was investigated. ARAE dose-dependently inhibited the active systemic anaphylaxis and serum IgE production induced by immunization with ovalbumin, Bordetella pertussis toxin and aluminum hydroxide gel. ARAE strongly inhibited IL-4-dependent IgE production by lipopolysaccharide-stimulated murine whole spleen cells. In the case of U266 human IgE-bearing B cells, ARAE also showed an inhibitory effect on the IgE production. These results suggest that ARAE has an anti-allergic activity by inhibition of IgE production from B cells.     

The introduction of one or two 3 beta-cholestanyl residues into benzylpenicilloyl-eicosa-L-lysines greatly potentiates their tolerogenicity for anti-benzylpenicillol IgE antibody formation

BALB/c mice were repeatedly immunized with microgram doses of benzylpenicilloylated Ascaris protein(s) (BPO9Asc) in alum. At different stages of the immune response, BPO21 eicosa-L-lysine or two analogs containing one or two hydrophobic p-oxymethylbenzyl-3 beta-cholestanyl succinate (OSuco) groups were injected. When injected early in the immune response, the anti-BPO IgE antibody formation was much more strongly and permanently suppressed by the lipophilic conjugates than by the hydrophilic BPO21 eicosa-L-lysine. A similar, but less marked, suppressive effect was observed on the anti-BPO IgG1 response. By adoptive cell transfer experiments, it was found that the OSuco-containing derivatives induce and act via suppressor T lymphocytes, since this cell-mediated suppression was sensitive to cyclophosphamide or to treatment with anti-Lyt-2.2 antibody plus complement. When these compounds were injected into repeatedly immunized mice producing late ongoing antibody responses no differences in suppression between hydrophilic and hydrophobic derivatives were observed. In this case, the IgE response was suppressed by about 50%, while the IgG1 response was not affected. These results are compatible with the suggestion that early IgE responses are most sensitive to T cell-mediated suppression and that T suppressor cells are better induced by lipophilic than by hydrophilic antigens. The late ongoing IgE response, on the other hand, is less amenable to T cell-induced suppression and tolerogenic effects brought about by plurivalent BPO antigens operate directly on hapten-specific IgE-bearing B cells, regardless of their lipophilic character.     

Identification of T cell subsets and class I and class II antigen expression in islet grafts and pancreatic islets of diabetic BioBreeding/Worcester rats

The BioBreeding/Worcester (BB/Wor) rat develops a spontaneous disorder that closely resembles human insulin-dependent (Type I) diabetes mellitus. The syndrome is preceded by lymphocytic insulitis that destroys pancreatic beta cells. The morphologic features of the spontaneous insulitis lesions are also observed within islets transplanted beneath the renal capsule of diabetes-prone and diabetic animals. This study reports the results of experiments in which immunohistochemical techniques were used to characterize the phenotype of the infiltrating mononuclear cells and detect the expression of class I and class II MHC antigens in native islets and islet transplants in diabetic and diabetes-prone BB/Wor rats. The infiltrates within native pancreatic islets and islet grafts were comprised predominantly of Ia+ cells (dendritic cells and macrophages) CD4+ cells (helper/inducer lymphocytes and macrophages), CD5+ (pan-T) cells and smaller numbers of CD8+ (cytotoxic/suppressor and NK) cells. Pancreatic and graft insulitis were accompanied by markedly enhanced class I antigen expression on islet and exocrine cells. Class II (Ia) antigens were not detected on normal islet cells, islets undergoing insulitis or on islet transplants subjected to immune attack. In islet grafts stained with polymorphic MAbs that distinguish Ia antigens of donor and host origin, Ia antigen expression was limited to infiltrating dendritic cells and macrophages of host origin. It is concluded that the phenotypes of infiltrating mononuclear cells that comprise the insulitis lesion in spontaneous BB/Wor diabetes, and the inflammatory attack on islets transplanted into diabetic BB/Wor rats are the same, that pancreatic islet and graft insulitis occur in the presence of enhanced class I antigen expression but in the absence of class II antigen expression, and that infiltrating Ia+ cells within islet grafts are exclusively of recipient (BB/Wor) origin and may explain the initiation of immune insulitis within grafts derived from donors of incompatible MHC.     

A 3 beta-cholestanyl-containing mono-benzylpenicilloyl oligoamide and peptide suppress anti-benzylpenicilloyl antibody formation in mice

A long-chain linear mono-benzylpenicilloyl (BPO) oligoamide and a succinoylated mono-BPO decalysine were tested in BALB/c mice for suppression of IgE and IgG1 antibody formation. Both compounds were available with either a free C-terminal end or were C-terminally linked to a hydrophobic 3 beta-cholestanyl residue. Only the sterol-containing derivatives suppressed hapten-specific IgE and IgG1 responses. Substantial suppression was obtained when the compounds were administered before primary or secondary, but not later immunizations. In an adoptive cell transfer experiment, spleen cells from tolerized animals actively suppressed anti-BPO IgE antibody formation of immune spleen cells. This effect was reversed by pretreatment of the tolerized spleen cells with anti-Lyt-2.2 antibody plus complement. The requirement for macrophages in the induction of T suppressor cells was demonstrated by injecting antigen-pulsed macrophages into naive recipients; upon immunization, only mice treated with tolerogen-pulsed macrophages showed suppressed anti-BPO IgE responses. It is suggested that lipid modification of antigens alters their processing and presentation by macrophages in a manner that leads to the induction of T suppressor cells. Injection of the cholestanyl derivatives into passively sensitized guinea pigs elicited anaphylactic reactions. By immune precipitation analysis and molecular weight estimation, these derivatives were shown to form micelles in aqueous solution. Therefore, the anaphylactic response appeared to be due to their behavior as multivalent antigens.