Inhibition Of IgE Binding Research Articles

Identification of Major Allergens in Watermelon

Background: Watermelon is a worldwide consumed Cucurbitaceae fruit that can elicit allergic reactions. However, the major allergens of watermelon are not known. The aim of this study is to identify and characterize major allergens in watermelon. Methods: Twenty-three patients allergic to watermelon took part in the study. The diagnosis was based on a history of symptoms and positive skin prick-prick tests to watermelon, confirmed by positive open oral challenge testing to watermelon pulp. Allergenic components were detected by SDS-PAGE and immunoblotting. Molecular characterization of IgE-binding bands was performed by N-terminal amino acid sequencing and mass spectrometry. Allergens were purified combining several chromatographic steps. Results: Several IgE binding bands (8–120 kDa) were detected in watermelon extract. Three major allergens were identified as malate dehydrogenase (36 kDa), triose phosphate isomerase (28 kDa) and profilin (13 kDa). Purified allergens individually inhibited IgE binding to the whole watermelon extract. Conclusions: All in all these results indicate that malate dehydrogenase, triose phosphate isomerase and profilin are major allergens involved in watermelon allergy.

Cross reactivity between European Hornet and Yellow Jacket venoms.

Summary Background: Cross-reactions between venoms may be responsible for multiple diagnostic positivities in hymenoptera allergy. There is limited data on the cross-reactivity between Vespula spp and Vespa crabro, which is an important cause of severe reactions in some parts of Europe. We studied by CAP-inhibition assays and immunoblotting the cross-reactivity between the two venoms. Methods: Sera from patients with non discriminative skin/CAP positivity to both Vespula and Vespa crabro were collected for the analyses. Inhibition assays were carried out with a CAP method, incubating the sera separately with both venoms and subsequently measuring the specific IgE to venoms themselves. Immunoblotting was performed on sera with ambiguous results at the CAP-inhibition. Results: Seventeen patients had a severe reaction after Vespa crabro sting and proved skin and CAP positive also to vespula. In 11/17 patients, Vespula venom completely inhibited IgE binding to VC venom, whereas VC venom inhibited binding to Vespula venom only partially (<75%). In 6 subjects the CAP-inhibition provided inconclusive results and their sera were analysed by immunoblotting. The SDS-PAGE identified hyaluronidase, phospholipase A1 and antigen 5 as the main proteins of the venoms. In 5 sera the levels of IgE against antigen 5 of Vespa crabro were higher than IgE against Vespula germanica, thus indicating a true sensitisation to crabro. Conclusion: In the case of multiple positivities to Vespa crabro and Vespula spp the CAP inhibition is helpful in detecting the cross-reactivities.

Allergy to Salsola Kali in a Salsola Incanescens-rich Area: Role of Extensive Cross Allergenicity

Pollens from the Salsola spp. are an important source of respiratory allergy in tropical countries. Our aim was to characterize the IgE binding proteins of S. incanescens pollen extract and study its cross-reactivity with S. kali pollen allergens. Prick tests with S. kali and S. incanescens pollen extracts were performed on eight respiratory allergy patients from Mashhad, Northeast Iran. The antigenic profiles and IgE-binding patterns of S. kali and S. incanescens pollen extracts were compared by SDS-PAGE and Western blotting, using individual sera from the salsola pollen-sensitive patients. Cross-reactivity of proteins in the two weeds was assessed by IgE- immunoblotting inhibition. S. kali and S. incanescens pollen extracts showed similar IgE-binding profiles in Western blotting. The IgE binding components of 39, 45, 66 and 85 kDa were detected in both pollen extracts. Furthermore, inhibition of the immunoblots revealed extensive inhibition of IgE binding to proteins and a close relationship between these two weeds allergens. S. incanescens pollen is a potent allergen source with several IgE binding components that shows a close allergenic relationship with S. kali. Our results suggest that in S. incanescens-rich areas, S. kali pollen extracts could be used as a diagnostic reagent for allergic patients to S. incanescens pollen.

Fine epitope mapping of humanized anti-IgE monoclonal antibody omalizumab

Omalizumab is a humanized anti-IgE antibody that inhibits IgE binding to its receptors on mast cells and basophils, thus blocking the IgE-mediated release of pharmacologic mediators from these cells. Previous studies have indicated that omalizumab binds to the Cε3 domain of IgE, which is the binding site of IgE receptors, but the precise epitope recognized by omalizumab is unknown. In this study, we employed the phage display peptide library technology to select peptides binding to omalizumab. A striking peptide sequence motif was recovered, which is homologous to the sequence 424HLP 426 within the Cε3 domain of IgE-Fc. Our results further indicated that omalizumab specifically bound to the synthesized peptide “ 421THPHLPRALMRS 432” containing the 424HLP 426 motif in IgE-Fc. We therefore conclude that the 424HLP 426 motif is the omalizumab epitope. This epitope overlaps with the high-affinity IgE receptor-binding site, thus providing insights into the structural basis for the mechanism of action of omalizumab.

Analysis of IgE binding proteins of Celtis tala pollen

Celtis tala is a tree, widely distributed in Argentina. Its significance as a source of allergic sensitisation is still unclear. Our aim was to partially characterise the IgE binding proteins of C. tala pollen. Skin prick tests using C. tala pollen extract were carried out in 25 patients with respiratory allergy. Celtis tala extract elicited positive skin reactions in five patients. Sensitised patients showed increased specific IgE levels to C. tala by immunodot. Immunoblotting assay demonstrated IgE binding to proteins of 50 kDa and 55 kDa. Celtis tala pollen extract is allergenically potent requiring approximately 50 ng of self-protein for 50% inhibition of IgE binding in enzyme_linked immunosorbent assay (ELISA) inhibition, whereas no cross-inhibition was detected with other types of pollen widely distributed in Argentina (Helianthus annuus and Prosopis spp.). Thus, these results taken together indicate that proteins of 50 kDa and 55 kDa may be considered as specific allergens of C. tala pollen.

A double‐blind placebo‐controlled birch allergy vaccination study II: correlation between inhibition of IgE binding, histamine release and facilitated allergen presentation

The pathogenesis of IgE-mediated allergic disease is closely related to the production of T-helper type 2 (Th2) cytokines, which lead to IgE production pivotal for activation of mast cells and basophils. Proliferating T cells along with eosinophils expanded and attracted by Th2 cytokines are major contributors to the late-phase reaction. The activation of these Th2 cells is strongly enhanced by CD23-mediated IgE facilitated allergen presentation (FAP). The present study aims to investigate the effect of specific immunotherapy (SIT)-induced allergen-specific non-IgE antibodies (blocking antibodies) on IgE binding to allergen, histamine release (HR) and CD23-mediated allergen uptake in antigen-presenting cells. Competition between IgE and non-IgE for allergen binding was studied by Advia Centaur antibody measurements, passively sensitized basophils were used to study HR and IgE-facilitated binding of allergen to B cells (FAP) was studied by flow cytometry. FAP measurements were performed both with and without the addition of a reference IgE serum, which was included to obtain optimal complex formation. The serum samples were obtained from birch pollen immunotherapy (n=21) or placebo control patients (n=21) before and after 1 and 2 years of treatment. Statistically significant reduction of all parameters investigated was observed after 1 year of treatment and the effect was maintained during the second year of treatment. There was a clear correlation between the two FAP measurements and between each of them and the level of T cell activation reported upon previously. Moreover, strong correlations were found between changes in FAP, IgE binding and HR. The present study clearly demonstrates that SIT induces changes in the composition of serum antibodies that inhibit IgE binding, HR and FAP to a similar extent. This suggests that these measurements, individually or in combination, may be used to monitor the immunological effect of SIT, even though direct correlations to changes in clinical parameters could not be demonstrated.

Aptameric green fluorescent protein (A‐GFP) selected from ribosome display (RD) library on solid phase IgE prevents human IgE binding to human FceRIa

Ribosome display is a complete in vitro system that permits selection of gene based on the physical association of its encoded message cognately to the protein it henceforth translates. The resulting ternary complexes of aptameric protein‐mRNA‐ribosome (ARM) specific for the ligand are immobilized on the designated solid phase ligand. Herein, green fluorescent protein (GFP) is fused to Cκ stuffer region as scaffold with 5′ T7 and Kozak/ATG sequences for transcription/translation‐coupled reactions. In non‐library modality, the presence of as low as 10 femtogram DNA rd4 construct of 1.2 kb, i.e., approximately 7,500 molecules, are sufficient for recursive rounds of selection by MAb anti‐GFP coated on solid phase. Next, ribosome display library constructed within site 6 and site 8/9 of GFP was employed for selecting aptameric protein that binds to human IgE on solid phase. An input 10 μg of 10‐mer aptameric GFP library for the respective dual site entails 7.5×1012 physical molecules (1.2 kb), expressed within a potential diversity of 1.0×1026 of possible peptide aptamers. Recombinant aptameric GFP was shown to bind to human IgE PS, BED, JW8, but not human IgG, lysozyme, BSA. Aptameric GFP also competes with the soluble receptor for binding human IgE on solid phase. Fragments of 25 to 100 bp from six IgE‐binding aptameric DNA by DNaseI were then shuffled, reassembled via internal homology by PCR. The molecularly evolved species exhibited improved binding to IgE. Thus, RD is an adequate technology platform for selecting aptameric GFP that specifically inhibits IgE binding to FcεRIα.

Allergy to human seminal fluid: Cross-reactivity with dog dander

Human seminal plasma (HSP) allergy is uncommon, with symptoms ranging from vulvovaginal pruritus to life-threatening anaphylaxis. Although several seminal plasma allergens have been reported and their molecular masses have been estimated to range between 12 and 75 kd, the prostate-specific antigen (PSA) has recently been identified as a causative allergen. Given that in a large number of cases symptoms appeared during or after the first intercourse, a cross-reactivity phenomenon might be implicated. We sought to assess the presence of IgE cross-reactivity among proteins from dog epithelium and HSP and to attempt to identify the allergens involved. Forty-one patients with dog epithelium allergy were selected. One of them experienced anaphylaxis in contact with her husband's seminal plasma. Skin prick tests, serum specific IgE measurements, SDS-PAGE immunoblotting, and inhibition tests were performed to study the pattern of IgE-binding proteins and the potential cross-reactivity between HSP and dog epithelium. Mass spectrometry was carried out to identify the protein involved in allergy reactions. Twenty-four percent of the sera from patients with dog epithelium allergy recognized an IgE-binding band of 28 kd in HSP immunoblotting. Mass spectrometry identified this band as the PSA. SDS-PAGE immunoblotting-inhibition showed a complete IgE-binding inhibition when sera from these patients were preincubated with dog dander extract. IgE cross-reactivity among proteins from dog dander and human PSA is demonstrated.

Characterization of a Hypoallergenic Recombinant Bet v 1 Variant as a Candidate for Allergen-Specific Immunotherapy

Background: Recombinant allergens and especially their hypoallergenic variants are promising candidates for a more effective and safer specific immunotherapy. Methods: Physicochemical and immunological characteristics of a folding variant of recombinant Bet v 1 (rBet v 1-FV) were investigated in comparison to natural Bet v 1 (nBet v 1) and the correctly folded recombinant Bet v 1 (rBet v 1-WT) by SDS-PAGE, size exclusion chromatography, multi-angle light scattering, circular dichroism, immunoblotting and enzyme allergosorbent test inhibition assay for detection of IgE reactivity and ELISA with Bet v 1-specific monoclonal antibodies. The functional IgE reactivity of the different Bet v 1 proteins was investigated using basophil activation in terms of CD203c expression and histamine release. T cell reactivity was investigated using T cell lines raised from birch pollen-allergic subjects against nBet v 1. Immunogenicity was investigated in mice. Results: Physicochemical characterization revealed purity, homogeneity and monomeric properties of rBet v 1-FV. Unlike nBet v 1 and rBet v 1-WT, rBet v 1-FV showed almost no IgE binding in immunoblots. The reduction of allergenicity was further proved by IgE-binding inhibition assays, basophil activation and histamine release. T cell reactivity was completely conserved, as demonstrated by proliferation of Bet v 1-specific T cell lines with multiple epitope specificities. rBet v 1-FV showed strong immunogenicity in mice. Conclusions: Due to its reduced IgE reactivity and decreased capacity to activate basophils, but retained T cell reactivity and strong immunogenicity, rBet v 1-FV proved to be a very promising candidate for specific immunotherapy in birch pollen-allergic subjects.

Kiwifruit allergy: actinidin is not a major allergen in the United Kingdom

Actinidin has previously been reported as the major allergen in kiwifruit. Objectives To investigate the relevance of actinidin in a well-characterized population of UK patients with kiwifruit allergy. To identify the allergens in kiwifruit, using Western blots, we examined the IgE-binding patterns of 76 patients with a history of kiwifruit allergy, 23 of who had had a positive double-blind, placebo-controlled food challenge. In addition, IgE binding to purified native actinidin was studied in 30 patients, and to acidic and basic isoforms of recombinant actinidin in five patients. Inhibition of IgE binding to kiwifruit protein extract by purified native actinidin was investigated by both inhibition immunoblots and inhibition ELISAs using pooled sera. Twelve protein bands in kiwifruit protein extract were bound by IgE. A protein band with a molecular weight of 38 kDa was the major allergen recognized by 59% of the population. IgE did not bind to actinidin in the kiwifruit protein extract, or to purified native or recombinant forms of actinidin during Western blotting. Pooled sera bound to kiwifruit protein extract but not purified actinidin on ELISA, and pre-incubating sera with actinidin did not inhibit IgE binding to kiwifruit protein extract on immunoblot or ELISA. A novel 38 kDa protein, not actinidin, is the major allergen in this large study population. Identification of major allergens in one patient group is therefore not necessarily reproducible in another; therefore, major allergens should not be defined until there is a sufficient body of data from diverse geographical and cultural populations.

Combination treatment with omalizumab and rush immunotherapy for ragweed-induced allergic rhinitis: Inhibition of IgE-facilitated allergen binding

The combination of anti-IgE (omalizumab) therapy with ragweed injection immunotherapy for seasonal allergic rhinitis results in a significant reduction in systemic side effects and enhanced efficacy compared with immunotherapy alone. One proposed mechanism of immunotherapy is to induce regulatory antibodies that inhibit facilitated antigen presentation. We sought to determine whether the combination protocol has a cumulative effect on inhibition of facilitated antigen presentation both during and after discontinuation of treatment. Ragweed allergen immunotherapy with and without omalizumab therapy was tested in a 4-arm, double-blind, placebo-controlled study. Flow cytometry was used to detect serum inhibitory activity for IgE-facilitated CD23-dependent allergen binding to B cells as a surrogate marker for facilitated antigen presentation. Serum ragweed-specific IgG4 was measured by means of ELISA. Immunotherapy alone resulted in partial inhibition of allergen-IgE binding after 5 to 19 weeks of treatment compared with baseline (P < .01). Complete inhibition of allergen-specific IgE binding was observed in both treatment groups receiving omalizumab (P < .001). Allergen-specific IgG4 levels were only increased after immunotherapy (P < .05), both in the presence and absence of anti-IgE treatment. Combined treatment resulted in the induction of long-lasting inhibitory antibody function for up to 42 weeks compared with either treatment alone. Ragweed immunotherapy induced serum regulatory antibodies that partially blocked binding of allergen-IgE complexes to B cells. Additional treatment with anti-IgE, by directly blocking IgE binding to CD23, completely inhibited allergen-IgE binding. The combination of ragweed immunotherapy and anti-IgE resulted in prolonged inhibition of allergen-IgE binding compared with either treatment alone, events that might contribute to enhanced efficacy.

Molecular Cloning and the Allergenic Characterization of Tropomyosin from Tyrophagus putrescentiae

Storage mites have been recognized as a cause of asthma and rhinitis. Studies from several countries have shown that the IgE-mediated allergy to storage mites is of considerable importance, especially in rural populations. This study aimed to identify and characterize new allergens from Tyrophagus putrescentiae. A partial cDNA sequence encoding tropomyosin was isolated from the cDNA library by immunoscreening using anti-mouse IgG1 sera raised against T. putrescentiae whole body extract. The deduced amino acid sequence shares 64-94% identity with previously known allergenic tropomyosins. Its recombinant protein was produced by using a pET 28b expression system and purified by affinity chromatography using Ni-NTA agarose. The IgE reactivities of tropomyosins from T. putrescentiae and Dermatophagoides farinae were compared by enzyme linked immunosorbent assay (ELISA). Recombinant Tyr p 10 showed 12.5% (5/40) IgE-binding reactivity, whereas recombinant Der f 10 showed 25% (10/40) IgE-binding reactivity against the same sera from storage mite-sensitized and house dust mite-sensitized subjects. Both recombinant Tyr p 10 and Der f 10 showed little inhibition of IgE binding to T. putrescentiae crude extract by ELISA. Tropomyosin seems to contribute only a small portion of the cross-reactivity with house dust mites.

Correlation Between SIT Induced Inhibition of IgE Binding, Histamine Release and Facilitated Allergen Presentation

Correlation Between SIT Induced Inhibition of IgE Binding, Histamine Release and Facilitated Allergen Presentation

Occupational respiratory allergy to roe deer

Although scattered reports have been published on roe deer allergenicity, no systematic studies of allergenicity or possible cross-reactions have appeared. To describe 2 patients with occupational roe deer allergy, demonstrated by positive skin and conjunctival provocation test results, and to note cross-reactions to other animal (mainly cow) allergens. Two workers at animal rehabilitation centers were sensitized to roe deer. One patient had a history of rhinoconjunctivitis and the other a history of rhinoconjunctivitis and probable asthma. Both patients underwent skin testing with a standard battery of inhaled and epithelial allergens and with roe deer hair and dander extract and conjunctival provocation tests with roe deer hair extract. Immunodetection for IgE (both patients) and IgE immunoblot inhibition tests to determine inhibitory effect (1 patient) were also performed. The results of skin tests and conjunctival provocation tests showed that both patients were sensitized to roe deer allergens. In one patient specific IgE to roe deer extract was detected, and this extract completely inhibited IgE binding to cow hair and dander extract in immunoblotting tests. Specific IgE to roe deer proteins could not be demonstrated in the other patient. Our results suggest that roe deer epidermal derivatives can cause occupational respiratory disease in exposed workers and that allergy to this species should be considered in individuals who present with similar symptoms and exposure histories. Immunoblot inhibition studies suggested the possibility of cross-reaction between roe deer proteins and cow proteins.

Evaluation of the sensitization rates and identification of IgE-binding components in wild and genetically modified potatoes in patients with allergic disorders

BackgroundThe potato is one of the most common types of genetically modified (GM) food. However, there are no published data evaluating the impact of genetic manipulations on the allergenicity of GM potatoes. To compare the allergenicity of GM potatoes with that of wild-type potatoes using in vivo and in vitro methods in adult allergy patients sensitized to potatoes.MethodsA total of 1886 patients with various allergic diseases and 38 healthy controls participated in the study. Skin-prick testing and IgE-ELISA were carried out with extracts prepared from wild-type and GM potatoes. An ELISA inhibition test was used to confirm the binding specificity. IgE-binding components in extracts from the two types of potato were identified by SDS-PAGE and IgE-immunoblotting. The effects of digestive enzymes and heat on the allergenicity of the extracts was evaluated by preincubating the potatoes with or without simulated gastric and intestinal fluids in the absence or presence of heat.ResultsPositive responses (ratio of the wheal size induced by the allergen to that induced by histamine (A/H) ≥ 2+) to wild-type or GM potato extracts, as demonstrated by the skin-prick test, were observed in 108 patients (5.7%). Serum-specific IgE was detected in 0–88% of subjects who tested positively. ELISA inhibition tests indicated significant inhibition when extract from each type of potato was added. IgE-immunoblot analysis demonstrated the presence of 14 IgE-binding components within the wild-type potato and 9 within the GM potato. Furthermore, a common 45-kDa binding component that yielded similar IgE-binding patterns was noted in more than 80% of the reactions using sera from patients sensitized to wild-type or GM potato. Exposure to simulated gastric fluid and heat treatment similarly inhibited IgE binding by extracts from wild-type and GM potatoes, whereas minimal changes were obtained following exposure of the extracts to simulated intestinal fluid.ConclusionOur results strongly suggest that genetic manipulation of potatoes does not increase their allergenic risk. The sensitization rate of adult allergy patients to both types of extract was 5.7%, and a common major allergen (45 kDa) was identified.

Structural Changes in the Lectin Domain of CD23, the Low-Affinity IgE Receptor, upon Calcium Binding

CD23, the low-affinity receptor for IgE (Fc epsilonRII), regulates IgE synthesis and also mediates IgE-dependent antigen transport and processing. CD23 is a unique Fc receptor belonging to the C-type lectin-like domain superfamily and binds IgE in an unusual, non-lectin-like manner, requiring calcium but not carbohydrate. We have solved the high-resolution crystal structures of the human CD23 lectin domain in the presence and absence of Ca2+. The crystal structures differ significantly from a previously determined NMR structure and show that calcium binding occurs at the principal binding site, but not at an auxiliary site that appears to be absent in human CD23. Conformational differences between the apo and Ca2+ bound structures suggest how IgE-Fc binding can be both calcium-dependent and carbohydrate-independent.

Analysis of IgE binding proteins of mesquite ( Prosopis juliflora) pollen and cross-reactivity with predominant tree pollens

Pollen from the mesquite tree, Prosopis juliflora, is an important source of respiratory allergy in tropical countries. Our aim was to partially characterize the IgE binding proteins of P. juliflora pollen extract and study cross-reactivity with prevalent tree pollen allergens. Intradermal tests with P. juliflora and five other tree pollen extracts were performed on respiratory allergy patients from Bikaner (arid) and Delhi (semi arid). Prosopis extract elicited positive skin reactions in 71/220 of the patients. Sera were collected from 38 of these 71 patients and all demonstrated elevated specific IgE to P. juliflora. Immunoblotting with pooled patients’ sera demonstrated 16 IgE binding components, with components of 24, 26, 29, 31, 35, 52, 58, 66 and 95 kDa recognized by more than 80% of individual patients’ sera. P. juliflora extract is allergenically potent requiring 73 ng of self-protein for 50% inhibition of IgE binding in ELISA inhibition. Cross-inhibition assays showed close relationship among P. juliflora, Ailanthus excelsa, Cassia siamea and Salvadora persica. IgE binding components of 14, 41, 52 and 66 kDa were shared allergens whereas 26 and 29 kDa were specific to P. juliflora. The findings suggest that purification of cross-reactive allergens will be helpful for diagnosis and immunotherapy of tree pollen allergic patients.

Occupational rhinoconjunctivitis and bronchial asthma due to Acalypha wilkesiana allergy

Acalypha wilkesiana, or copperleaf, is a plant of the Euphorbiaceae family. Although it is widely known as an outdoor ornamental plant, no cases of A. wilkesiana allergy have been reported to date. To describe a patient with occupational respiratory allergy to A. wilkesiana. Extracts from A. wilkesiana leaves and flowers were used for skin prick testing, specific conjunctival and bronchial challenge tests, and in vitro studies. These studies range from A. wilkesiana specific IgE determination to sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunodetection of A. wilkesiana protein bands in patient serum samples and immunoblot inhibition by preincubation with Salsola kali and Chenopodium album pollen extracts. Our patient had positive skin prick test reactions to A. wilkesiana leaf and flower extracts; negative reactions were found in a control group of 20 atopic patients. On immunodetection of A. wilkesiana extracts in patient serum samples, as many as 9 different IgE-binding proteins, with apparent molecular weights of 16 to 86 kDa, were revealed. Preincubation with S. kali and C. album pollen extracts completely inhibited IgE binding to the A. wilkesiana extract. Specific bronchial challenge resulted in a spirometric 30% decline in forced expiratory volume in 1 second with respect to baseline 1 minute after 1:100 (vol/vol) A. wilkesiana extract solution inhalation; 2 atopic controls had negative bronchial challenge test results. Acalypha wilkesiana is a new etiologic agent for IgE-mediated occupational respiratory allergy.

IgE cross-reactivity between the major peanut allergen Ara h 2 and tree nut allergens

Allergy to peanut and tree nuts is characterised by a high frequency of life-threatening anaphylactic reactions and typically lifelong persistence. Although peanut is the most common cause of nut allergy, peanut allergic patients are frequently also sensitive to tree nuts. It is not known if this is due to cross-reactivity between peanut and tree nut allergens. In this study, the major peanut allergen Ara h 2 was cloned from peanut cDNA, expressed in E. coli cells as a His-tag fusion protein and purified using a Ni-NTA column. Immunoblotting, ELISA and basophil activation indicated by CD63 expression all confirmed the IgE reactivity and biological activity of rAra h 2. To determine whether or not this allergen plays a role in IgE cross-reactivity between peanut and tree nuts, inhibition ELISA was performed. Pre-incubation of serum from peanut allergic patients with increasing concentrations of almond or Brazil nut extract inhibited IgE binding to rAra h 2. Purified rAra h 2-specific serum IgE antibodies also bound to proteins present in almond and Brazil nut extracts by immunoblotting. This indicates that the major peanut allergen, Ara h 2, shares common IgE-binding epitopes with almond and Brazil nut allergens, which may contribute to the high incidence of tree nut sensitisation in peanut allergic individuals.

Cloning and Characterization of Walnut Lipid Transfer Protein: Not a Major Allergen in Walnut-Allergic U.S. Patients

RATIONALE: Lipid transfer protein, a protein with inherent resistance to gastric digestion, has been isolated from walnut kernel and determined to bind IgE from Italian patients with walnut allergy. Many of these patients had been initially sensitized to lipid transfer protein in peaches. We sought to determine if lipid transfer protein was also a major allergen for U.S. patients with severe walnut allergy, and if the cloned, expressed protein could inhibit IgE binding to the native protein.