IgE antibody-specific activity in human allergic disease

IgE antibody concentration, affinity, clonality and specific activity (also known as the allergen-specific IgE to total IgE ratio) influence the translation of IgE responses into clinically evident allergic symptoms following allergen exposure. Reported IgE-specific activity levels >3–4% place allergic individuals undergoing anti-IgE (Omalizumab^®) therapy at a disadvantage for poor resolution of their allergy symptoms following manufacturer’s recommended dosing schemes. We investigated the hypothesis that the specific activity of the IgE antibody response is highly variable with respect to age, allergen specificity and an individual’s total serum IgE level. Second, we investigated whether the IgE-specific activity level influences the extent and rate of loss of effector cell mediator release. IgE-specific activity distributions were plotted against age, allergen specificity and total serum IgE using 18,950 paired total IgE and allergen-specific IgE antibody data obtained from the analysis of sera from 3,614 allergic subjects and covering 182 allergen specificities. The fraction of specific IgE antibody of the total serum IgE was dependent on age of the individual, epitope specificity (clonality) and total serum IgE. The youngest group of allergic individuals with the lowest total serum IgE levels tended to have the highest allergen-specific IgE to total IgE ratios. Hymenoptera venom (54%), peanut (33%) and milk (27%) were the three allergen specificities that elicited the highest frequency of IgE-specific activities >4% among sensitized individuals. A prospective double-blind, placebo-controlled clinical study involving anti-IgE treatment of cat-allergic subjects showed IgE-specific activity was remarkably constant over the 16-week course of treatment, despite the up to 8-fold rise in total serum IgE following repetitive Omalizumab^® administration. Changes in specific and total IgE levels paralleled each other in patients receiving anti-IgE therapy. The fastest rate of reduction in cat allergen-induced basophil histamine release following anti-IgE therapy was observed when the cat-specific IgE to total IgE ratio was <2.5%. This reflected the more rapid loss of surface cat-specific IgE antibody with anti-IgE therapy in allergic individuals who displayed a more diverse IgE antibody repertoire. We conclude that IgE-specific activity is an age-, IgE heterogeneity- and total serum IgE-dependent variable that influences the magnitude of effector cell mediator release, and by inference, ultimate allergic symptom induction.     

A monoclonal antibody specific for a carbohydrate epitope recognizes an IgE-binding determinant shared by taxonomically unrelated allergenic pollens

Carbohydrate epitopes are capable of binding human IgE from allergic subjects and these epitopes play a role in the cross-reactivity between allergens from unrelated sources. A monoclonal antibody (5E6), specific for a carbohydrate epitope detectable on components of Cupressus arizonica pollen extract, has been produced and characterized. To study the relationship between the epitopes recognized by the monoclonal antibody and by IgE from allergic subjects. To investigate the presence of such carbohydrate IgE determinant in extracts from 21 pollen species belonging to 16 taxonomically related and unrelated families, by means of the monoclonal antibody. IgG-depleted fraction from protein G-purified human allergic serum was obtained. The monoclonal antibody and the IgE from the purified fraction were tested on two glycoproteins, polyamine oxidase and ascorbate oxidase, adsorbed on the ELISA plates. The relationship between the monoclonal- and the IgE-recognized epitopes was investigated by ELISA-competition experiments. Analysis of the distribution of this carbohydrate epitope was performed by direct binding of the monoclonal antibody onto the various extracts. The monoclonal antibody and the IgE were able to bind carbohydrate epitopes on the two plant glycoproteins, ascorbate oxidase and polyamine oxidase. Polyamine oxidase shows only one N-glycosilation site whose carbohydrate moiety seems to be composed of a branched chain of seven ordered sugars, i.e. two N-acetyl-D-glucosamine-, three mannose-, one fucose- and one xylose-residues. This structure bears the epitope recognized by mAb 5E6. Human IgE from the IgG-depleted fraction were found capable of inhibiting the monoclonal antibody binding. The allergenic epitope identified was shared by a large number of extracts with different levels of reactivity (OD490 ranging from 0.110 to 2.060). Our data support the finding that a monoclonal antibody specific for a carbohydrate epitope of Cupressus arizonica pollen extract detects an epitope which is also recognized by IgE from allergic subjects. This characterized reagent could be a useful tool for studying distribution of cross-reactive carbohydrate determinants in allergenic pollen extracts and their components.     

Interference of carbamazepine and carbamazepine 10,11-epoxide in the fluorescence polarization immunoassay for tricyclic antidepressants: estimation of the true tricyclic antidepressant concentration in the presence of carbamazepine using a mathematical model

We evaluated effects of carbamazepine and its metabolite, carbamazepine 10,11-epoxide, on the measurement of tricyclic antidepressant (TCA) concentrations in serum using the fluorescence polarization immunoassay (FPIA). We determined apparent TCA concentrations in 30 patients who were receiving carbamazepine but no TCAs. Carbamazepine concentrations ranged from 1.4 to 20.9 microg/mL (5.9-88.4 micromol/L); the observed apparent TCA concentrations ranged from 31.8 to 130.1 ng/mL (113.4-463.9 micromol/L). When aliquots of the drug-free serum pool were supplemented with known concentrations of carbamazepine or its metabolite, we observed significant apparent TCA concentrations using the FPIA; however, interference of carbamazepine was more than 3-fold more than its metabolite. When serum pools prepared from patients receiving TCA but no anticonvulsant medications were supplemented with known amounts of carbamazepine or its metabolite, we observed falsely elevated TCA concentrations. We formulated an equation to calculate the apparent TCA concentration from known carbamazepine concentrations. If carbamazepine and TCAs are present in a specimen, the true TCA concentration can be estimated by subtracting the calculated TCA concentration (due to carbamazepine) from the observed TCA concentration as measured by the TCA FPIA. This mathematical modeling is feasible because TCAs, even at very high concentrations, showed no interference with the carbamazepine FPIA.     

Effects of carbamazepine on serum electrolytes in affectively ill patients

The effects on serum electrolytes of carbamazepine, an acute and prophylactic treatment for manic-depressive illness, were assessed in subjects with primary affective disorder. Carbamazepine caused statistically significant, but clinically insubstantial, reductions in serum sodium and calcium, but not in the other electrolytes measured. Decreases in serum sodium and calcium were not related to carbamazepine dose, blood levels, or the degree of clinical improvement. The theoretical implications of these findings are discussed.     

Lymphocyte-stimulation tests and patch tests to carbamazepine hypersensitivity.

Seven cases of severe hypersensitivity to carbamazepine (Tegretol) were described in patients with epilepsy or trigeminal neuralgia. Clinical manifestations consisted of fever, rash, facial oedema, lymphadenopathy, impaired liver function, eosinophilia and atypical lymphocytes in the peripheral blood. Lymphocyte-stimulation tests with carbamazepine in vitro showed positive results in all cases; patch tests with carbamazepine were positive in six cases. In two cases the lymphocyte-stimulation tests with carbamazepine were found to be negative during, and shortly after, the illness. However, when the tests were repeated several months later, they turned out to be positive. Lymphocyte reactivity to PPD and PHA in vitro was also impaired during the acute phase of the disease. Thus false-negative lymphocyte-stimulation tests may be found in the first months following such a hypersensitivity reaction, probably due to impaired lymphocyte reactivity. As carbamazepine is a potent drug and is often prescribed for long periods together with other anticonvulsants, it seems important to prove that the allergic reaction is caused by carbamazepine. If the lymphocyte-stimulation test in vitro or the patch test with carbamazepine is found to be negative during or shortly after the illness, they should be repeated several months later.     

Hyponatraemia during low-dose carbamazepine therapy

We report the syndrome of inappropriate antidiuresis as a much earlier side-effect of carbamazepine administration in a 29-year Nigerian female patient with generalized tonic-elonic seizures. Although asymptomatic, the biochemical abnormality improved after discontinuation of carbamazepine. Hyponatraemia developed after rechallenge with controlled release carbamazepine. The authors suggest that serum sodium levels be carried out before commencement of carbamazepine and caution be used in prescribing carbamazepine to patients with low or borderline low sodium values.     

Clinical correlations between long-term (IgE) and short-term (IgG S-TS) anaphylactic antibodies in atopic and 'non-atopic' subjects with respiratory allergic disease

The sera of atopic and non-atopic persons with allergic pulmonary disorders were examined for long-term sensitizing, IgE and short-term sensitizing heat-stable (S-TS) antibodies which were present separately or together in the sera of some patients sensitive to antigens such as budgerigar serum proteins and Aspergillus fumigatus. In fourteen atopic patients with extrinsic asthma, six had both types of antibody to common allergens, and of nine non-atopic patients with cryptogenic (intrinsic) asthma, four had only heat-stable short-term sensitizing antibodies. The sera of atopic subjects with type I prick test reactions and positive RAST's, showed specific IgE antibody by baboon PCA tests to budgerigar serum proteins, A. fumigatus, Timothy grass pollen extract and hen egg extract, and not to Dermatophagoides farinae, possibly because of naturally occurring mite antibodies in the baboon. The sera of non-atopic asthmatics, who had given negative prick test but positive immediate, dual or late intracutaneous tests, and only late asthmatic reactions, contained precipitins in most cases and gave little or no RAST reaction. On baboon PCA these sera contained either, S-TS antibody alone, or S-TS plus long-term sensitizing antibody, or long-term sensitizing antibody alone. Some of the sera with long-term sensitizing antibody contained blocking antibody which could diffuse away in the 24 hr delay for the baboon PCA test and could also be responsible for the negative RAST. Tests with insoluble anti-IgE immuno-adsorbents on two sera from persons sensitive to aspergillus confirmed that the S-TS activity was not due to IgE, and on two sera with negative RAST and negative prick tests to budgerigar serum antigens confirmed that the 24 hr monkey PCA responses were due to IgE.     

Activation of T cells by carbamazepine and carbamazepine metabolites

BACKGROUND: T-cell-mediated hypersensitivity is a rare but serious manifestation of drug therapy. OBJECTIVES: To explore the mechanisms of drug presentation to T cells and the possibility that generation of metabolite-specific T cells may provoke cross-sensitization between drugs. METHODS: A lymphocyte transformation test was performed on 13 hypersensitive patients with carbamazepine, oxcarbazepine, and carbamazepine metabolites. Serial dilution experiments were performed to generate drug (metabolite)-specific T-cell clones to explore the structural basis of the T-cell response and mechanisms of antigen presentation. 3-Dimensional energy-minimized structures were generated by using computer modeling. The role of drug metabolism was analyzed with 1-aminobenzotriazole. RESULTS: Lymphocytes and T-cell clones proliferated with carbamazepine, oxcarbazepine, and some (carbamazepine 10,11 epoxide, 10-hydroxy carbamazepine) but not all stable carbamazepine metabolites. Structure activity studies using 29 carbamazepine (metabolite)-specific T-cell clones revealed 4 patterns of drug recognition, which could be explained by generation of preferred 3-dimensional structural conformations. T cells were stimulated by carbamazepine (metabolites) bound directly to MHC in the absence of processing. The activation threshold for T-cell proliferation varied between 5 minutes and 4 hours. 1-Aminobenzotriazole, which inhibits cytochrome P450 activity, did not prevent carbamazepine-related T-cell proliferation. Substitution of the terminal amine residue of carbamazepine with a methyl group diminished T-cell proliferation. CONCLUSION: These data show that carbamazepine and certain stable carbamazepine metabolites stimulate T cells rapidly via a direct interaction with MHC and specific T-cell receptors. CLINICAL IMPLICATIONS: Some patients with a history of carbamazepine hypersensitivity possess T cells that cross-react with oxcarbazepine, providing a rationale for cross-sensitivity between the 2 drugs.     

Can alternative epitope mapping approaches increase the impact of B‐cell epitopes in food allergy diagnostics?

In vitro allergy diagnostics are currently based on the detection of specific IgE binding on intact allergens or a mixture thereof. This approach has drawbacks as it may yield false‐negative and/or false‐positive results. Thus, we reviewed the impact of known B‐cell epitopes of food allergens to predict transience or persistence, tolerance or allergy and the severity of an allergic reaction and to examine new epitope mapping strategies meant to improve serum‐based allergy diagnostics. Recent epitope mapping approaches have been worthwhile in epitope identification and may increase the specificity of allergy diagnostics by using epitopes predominately recognized by allergic patients in some cases. However, these approaches did not lead to discrimination between clinically relevant and irrelevant epitopes so far, since the polyclonal serum IgE‐binding epitope spectrum seems to be too individual, independent of the disease status of the patients. New epitope mapping strategies are necessary to overcome these obstacles. The use of patient‐derived monoclonal antibodies instead of patient sera for functional characterization of clinically relevant and irrelevant epitope combinations, distinguished by their ability to induce degranulation, might be a promising approach to gain more insight into the allergic reaction and to improve serum‐based allergy diagnostics.     

Serum vascular endothelial growth factor in allergic rhinitis and systemic lupus erythematosus

Allergic rhinitis (AR) is characterized by an inflammatory reaction sustained by Th2 polarization, whereas systemic lupus erythematosus (SLE) is a typical autoimmune disorder. Vascular endothelial growth factor (VEGF) is a critical mediator of inflammation. The aim of this study was to compare serum VEGF levels in three groups of subjects: 40 normal subjects, 40 allergic patients, evaluated before and after specific immunotherapy, and 40 patients with inactive SLE. Patients who were allergic before immunotherapy had the lowest VEGF serum levels, which significantly increased after treatment; SLE patients had the highest VEGF serum levels. This comparative study provides evidence that serum VEGF levels depend on the type of immune response: they are high in autoimmune disease and low in Th2-polarized allergic reaction. The relevance of this phenomenon is further apparent, as it is also observed in patients with inactive disease.     

Hemoperfusion study with carbamazepine in vitro

Sorption efficacy of active charcoal, Amberlite XAD-2 and Amberlite XAD-4 in hemoperfusion with carbamazepine was investigated in vitro. Authors carried out 15 hemoperfusions with carbamazepine in a closed system using the above mentioned sorbents; The perfusion solution used was 3,000 ml of 0.9% NaCl. The carbamazepine concentration in the solution was 52.5+/-3.0 mg/L. In addition, two in vitro hemoperfusions with carbamazepine using Amberlite XAD-2 sorbent and 1.500 ml of human plasma as a perfusion solution, were also performed. The concentration of carbamazepine in plasma was 50.4+/-1.2 mg/L. Individual hemoperfusions lasted 5 hours. Zero values of carbamazepine were achieved after 120 min of Amberlite XAD-2 or Amberlite XAD-4 hemoperfusion in 0.9% NaCl and after 240 min of hemoperfusion in human plasma. The results gathered suggest that the most effective in vitro hemoperfusion with carbamazepine was obtained with Amberlite XAD-2 or Amberlite XAD-4 and the least effective was with active charcoal. The authors recommend the use of hemoperfusion with Amberlite XAD-2 or Amberlite XAD-4 for acute intoxications with carbamazepine in humans. In addition, authors successfully performed 5-hour hemoperfusion using Amberlite XAD-4 in vivo, in an 18-year-old girl, who suffered from acute carbamazepine intoxication after a suicidal attempt. This treatment led to the rapid survival.     

抗小鼠TLR2胞外段单表位抗体TSP-2对OVA诱导的小鼠过敏性哮喘气道炎症的影响

目的观察抗小鼠TLR2胞外段单表位（mTLR2ECD）抗体（TSP-2）对OVA诱导的小鼠过敏性哮喘气道炎症的影响。方法用肺泡灌洗、组织切片及特异染色、免疫组化、ELISA法检测TSP-2对小鼠哮喘模型的气道炎症和炎症细胞的浸润以及肺组织中肺泡及支气管结构的影响。结果发现静脉给予抗体TSP-2能有效抑制气道炎症和炎症细胞的浸润。主要表现在肺泡灌洗液中的白细胞浸润减少、减轻血清中OVA特异性IgE抗体的降低以及抑制OVA诱导的小鼠肺泡毛细血管充血水肿等病理变化。结论抗体TSP-2对过敏性哮喘的病理变化有一定的抑制作用。     

Stevens-Johnson syndrome due to carbamazepine

Two patients with psychotic disorders who developed Stevens-Johnson Syndrome while on treatment with carbamazepine is reported due to its rarity. Dermatological side-effects of carbamazepine may be more common in psychiatric as compared to neurological patients.     

Bronchial histamine reactivity: its relationship to the reactivity of the bronchi to allergens

The aim of this study was to examine the proposal that the magnitude of the response of the bronchi to an immediate allergic reaction depends not only on the degree of sensitization of the bronchi by allergen specific IgE antibody but also on the reactivity of the bronchi to the vasoactive mediators which are released during immediate allergic reactions. This was done by determining the bronchial reactivity to Dermatophagoides pteronyssinus and to histamine of both symptomatic and asymptomatic groups of atopic subjects who had comparable serum levels of D. pteronyssinus specific IgE. Positive bronchial responses to the D. pteronyssinus extract were recorded with both the symptomatic and asymptomatic subjects, the mean bronchial threshold dose of allergen being significantly higher in the asymptomatic than in the asthmatic patients. There was a highly significant correlation between the serum level of allergen specific IgE and the bronchial threshold dose of allergen extract and also between the bronchial threshold dose of allergen extract and of histamine in all groups of subjects. The ability to predict bronchial reactivity to the allergen from the serum level of allergen specific IgE within each group was significantly better if the bronchial reactivity to histamine was included in the correlation analysis. This supports the hypothesis that whether a particular subject who is producing specific IgE antibody will develop symptoms on the inhalation of that allergen depends not only on the amount of allergen which he inhales and on the degree of sensitization of his bronchi but also on the reactivity of his bronchi to the vasoactive mediators which are released by allergen–IgE interaction.     

Reactions during hemodialysis caused by allergy to ethylene oxide gas sterilization

In patients receiving long-term hemodialysis (HD), we have examined the presence of IgE-dependent sensitization to ethylene oxide (EO) gas, which is used for sterilization of disposable medical products including dialyzers. Serum was obtained from 25 patients who experienced acute allergic reactions during HD, five patients receiving HD with isolated eosinophilia, and 37 unselected patients receiving HD. Sera from 22 of 25 of the allergic reaction group and from five of 35 of the unselected group were demonstrated to contain IgE antibodies with specificity for EO. Corresponding IgG antibodies were also present. No such antibodies were detected in serum from normal controls or ragweed-allergic patients. The serum from one patient with isolated eosinophilia had a borderline elevated IgE antibody level. These results demonstrate a close relationship between the presence of IgE antibodies to EO and HD-related allergic reactions in this patient population.     

Assessing basophil functional measures during monoclonal anti-IgE therapy

Assessing the impact of therapeutic interventions on the clinical and immunologic responses of allergic subjects is a topic of extensive investigation. Available approaches include the measurement of in vivo allergen challenge responses, serologic measures, or in vitro studies of cells that participate in the allergic reaction. Several decades of work support that measures of allergen responses of IgE-bearing peripheral blood basophils can reflect clinical expression of allergic disease. In the last decade, an immune-based therapy targeting IgE, omalizumab, has emerged as an adjunct treatment for a variety of allergic diseases. This monoclonal humanized IgG antibody specifically binds circulating IgE at a region in the Fc tail that prevents IgE attachment to high affinity IgE receptor (FcεRI) bearing cell types such as tissue mast cells and blood basophils. This review focuses on methods to monitor changes of basophil allergen reactivity with a focus on omalizumab therapy and the implications for clinical disease management.     

Pure red cell aplasia associated with carbamazepine

Therapeutic indications for carbamazepine have grown in recent years and more and more patients are taking this drug for various neurological and psychiatric disorders. Though haematological complications like aplastic anaemia etc., due to carbamazepine therapy are well known pure red cell aplasia is exceedingly rare. A 33 year old man was diagnosed to have pure red cell aplasia with positive Coomb's test following 5 months of carbamazepine therapy. He recovered completely. Carbamazepine induced pure red cell aplasia is a reversible condition if recognised early.     

In vitro lymphocyte proliferation by carbamazepine, carbamazepine-10, 11-epoxide, and oxcarbazepine in the diagnosis of drug-induced hypersensitivity

In vitro lymphocyte proliferation induced by carbamazepine (CBZ) was evaluated in nine patients with hypersensitivity to this drug. Lymphocytes from all hypersensitive patients responded by significantly enhanced DNA synthesis to CBZ when patients were compared with 33 tested control subjects. However, lymphocytes from five of six hypersensitive patients were not stimulated by carbamazepine-10, 11-epoxide, and oxcarbazepine (OXC), and this was confirmed clinically in two CBZ hypersensitive patients with OXC therapy. The results indicate that OXC may be suitable as an alternative therapy in some patients with CBZ hypersensitivity.     

Depression of immune competence by phenytoin and carbamazepine. Studies in vivo and in vitro.

Depression of one or more parameters of cellular and/or humoral immune responses was found in 60% of general hospital patients treated with phenytoin and 47% of patients treated with carbamazepine. Phenytoin-treated patients failed to manifest delayed hypersensitivity (DHS) reactions to common antigens, and to make antibody to Salmonella typhi and tetanus toxoid. Serum levels of IgA and IgM, DNA synthesis in circulating leucocytes, and phytohaemagglutinin (PHA) induced deoxyribonucleic acid synthesis were also low. Depression of IgA, DHS reactivity and antibody responsiveness to S. typhi were shown to develop after the commencement of phenytoin therapy in a study of eleven patients. The presence of immunological defects was independent of the dosage of drug, its serum concentration, the duration of therapy and the sex of the subject. Studies in vitro provided evidence that immunosuppression was the result of a direct effect of phenytoin on the metabolism of lymphoid cells. Carbamazepine was shown to have a similar but less potent direct effect. Pharmacological concentrations of phenytoin caused a significant depression of DNA synthesis in PHA-stimulated and non-stimulated blood cell cultures in vitro. High concentrations in addition caused depression of cell counts, lymphocyte blastogenesis, ribonucleic acid and protein synthesis. Phenytoin was not cytocidal at concentrations of up to 125 mug/ml. Depression of DNA synthesis by phenytoin was maximal when phenytoin was added within 4-8 hr of the addition of PHA. PHA-induced DNA synthesis was not significantly affected by pre-incubation with phenytoin. In vivo, the presence of immunological defects was not related to phenytoin-induced folic acid deficiency. High concentrations of carbamazepine, but not phenobarbitone or diazepam caused a significant depression of PHA-stimulated DNA synthesis in blood cell cultures. The data show that immunosuppression is a common side-effect of phenytoin therapy, and that lymphoma is rare. They suggest that in the presence of phenytoin-induced immunosuppression another factor, or factors are required to induce the formation of lymphoma.     

Skin testing and extrinsic allergic alveolitis

Skin testing with six common allergens, tuberculin and a sterile avian antigen preparation from pigeon serum was performed on 102 pigeon fanciers. The incidence of positive prick tests to common allergens was no different for subjects with extrinsic allergic alveolitis. EAA, caused by avian exposure than the whole group. Positive immediate weal and flare reactions following skin prick testing with avian antigen occurred in 22 subjects and was closely correlated with atopy. However, when the same antigen was administered intradermally, 69 subjects developed an immediate (15 min) weal and flare reaction which did not correlate with atopy, instead, the weal diameter correlated significantly with the serum IgG antibody titre against pigeon serum gamma-globulin antigen, and furthermore, the higher grades of reaction were highly selective for subjects with EAA. Ten subjects, all with strong early intradermal skin reactions, developed a late (4-6 h) skin reaction; this was again highly selective for EAA. The subjects with cutaneous anergy to tuberculin had markedly higher IgG antibody titres to avian antigens, and these included the majority of the subjects with alveolitis.