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1.
J Investig Allergol Clin Immunol ; 32(3): 206-212, 2022 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-33830040

RESUMO

OBJECTIVES: To determine the usefulness of the in vitro and in vivo methods used in the diagnosis of kiwifruit allergy and to specifically assess the impact of seed proteins on sensitivity. METHODS: We performed skin prick tests (SPTs) using various commercial extracts, homemade pulp, and seed extracts and prick-prick tests with kiwifruit on 36 allergic patients. The presence of specific IgE (sIgE) was assessed using the ImmunoCAP (kiwifruit extract), ELISA (Act d 1, Act d 2), ISAC, and FABER assays. Immunoblotting of seed extract was carried out, and a single-blind oral food challenge was performed with whole seeds in seed-sensitized individuals. RESULTS: The prick prick test with kiwifruit demonstrated the highest diagnostic capacity (81.8% sensitivity and 94.1% specificity) among the in vivo tests. The sIgE levels measured using ImmunoCAP (kiwifruit extract) showed a similar sensitivity to that of global ISAC and FABER (63.9%, 59.5%, and 58.3%, respectively). Act d 1 was the major allergen. Sensitization to Act d 1 was associated with positive sIgE results to whole kiwifruit extract detected by ImmunoCAP (P<.000). A positive SPT result to kiwifruit seeds was associated with severe symptoms induced by kiwifruit (P=.019) as a marker of advanced disease, but not with clinically relevant sensitization. Challenge testing with kiwifruit seeds performed on 8 seed-sensitized patients yielded negative results. CONCLUSION: Sensitization to Act d 1 is associated with a positive result in conventional diagnostic techniques, whereas kiwifruit seed sensitization does not increase the sensitivity of the diagnostic techniques evaluated.


Assuntos
Actinidia , Hipersensibilidade , Actinidia/efeitos adversos , Alérgenos , Testes Diagnósticos de Rotina , Humanos , Imunoglobulina E , Extratos Vegetais , Método Simples-Cego , Testes Cutâneos/métodos
2.
Sens Actuators B Chem ; 345: 130394, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34248283

RESUMO

The standard rapid approach for the diagnosis of coronavirus disease 2019 (COVID-19) is the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. The detection of specific anti-SARS-CoV-2 immunoglobulins is crucial for screening people who have been exposed to the virus, whether or not they presented symptoms. Recent publications report different methods for the detection of specific IgGs, IgMs, and IgAs against SARS-CoV-2; these methods mainly detect immunoglobulins in the serum using conventional techniques such as rapid lateral flow tests or enzyme-linked immunosorbent assay (ELISA). In this article, we report the production of recombinant SARS-CoV-2 spike protein and the development of a rapid, reliable, cost-effective test, capable of detecting immunoglobulins in serum and saliva samples. This method is based on interferometric optical detection. The results obtained using this method and those obtained using ELISA were compared. Owing to its low cost and simplicity, this test can be used periodically for the early detection, surveillance, detection of immunity, and control of the spread of COVID-19.

3.
Artigo em Inglês | MEDLINE | ID: mdl-28760720

RESUMO

Profilin is a protein that is present in all eukaryotic cells and is responsible for cross-reactivity between pollen, latex, and plant foods. It has been classically acknowledged as a minor or nearly irrelevant allergen, although recent data are changing this conception. The objective of this manuscript is to provide a comprehensive review of published data on the role of this ubiquitous allergen in pollen, latex, and plant food allergy. The patterns of recognition of this minor allergen follow a north-south gradient. Although present in all pollens and vegetables, profilin is significantly associated with allergy to grass pollen and to Cucurbitaceae fruits. Heb v 8, the latex profilin, is usually a marker of profilin allergy in plant food-allergic patients, although it has no clinical relevance in latex allergy. Sensitization to profilin jeopardizes the diagnosis of pollen allergy and selection of immunotherapy, and although component-resolved diagnosis can identify its impact, there are no tailored treatments available. In recent years, several new publications have shown how profilin should be taken into account and, under certain circumstances, considered a marker of severity, an allergen capable of inducing respiratory symptoms, and, in its natural purified form, a potential candidate for etiological treatment of food allergy. Current data on profilin strongly support the need for a shift in the previously accepted paradigm for this allergen. More research should be done to assess the real clinical impact of sensitization in specific populations and to develop therapeutic strategies.


Assuntos
Profilinas/imunologia , Alérgenos/imunologia , Hipersensibilidade Alimentar/imunologia , Humanos , Látex/imunologia , Hipersensibilidade ao Látex/imunologia , Proteínas de Plantas/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia
4.
Clin Exp Allergy ; 47(3): 339-350, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28160513

RESUMO

BACKGROUND: The peach non-specific lipid transfer protein, Pru p 3, is the primary sensitizer in fruits and responsible for severe reactions in the Mediterranean area. Peach allergy is frequently associated with other allergies such as peanut. Therefore, it is important to assess how specific immunotherapy to Pru p 3 could affect both peach and peanut tolerance. OBJECTIVES: To evaluate peach and peanut desensitization and immunological changes after 1 year of Pru p 3 sublingual immunotherapy (SLIT) in patients with systemic allergic reactions to peach and/or peanut. METHODS: Forty-eight peach allergic patients, 36 treated with SLIT and 12 non-treated, were monitored for 12 months. Treated patients were subclassified as peanut allergic (Group A), sensitized (Group B) or tolerant (Group C). SLIT effect was evaluated by skin prick test (SPT) reactivity and food challenge. Immunological changes were evaluated by monitoring sIgE and sIgG4 levels and basophil reactivity. RESULTS: After 1 year of SLIT, the weal area in SPT significantly decreased and a significant increase in peach threshold in treated patients was observed (P < 0.001). Patients in Group A showed a significant decrease in peanut SPT weal area and an increase in peanut threshold (P < 0.001). Immunological changes were observed in treated patients only, with a significant decrease in sIgE and a parallel increase in sIgG4, sIgG4/sIgE and basophil reactivity for both Pru p 3 and Ara h 9. CONCLUSIONS AND CLINICAL RELEVANCE: After 1 year, Pru p 3 SLIT induces both desensitization and immunological changes not only for peach but also for other food allergens relevant in the induction of severe reactions such as peanut.


Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Arachis/efeitos adversos , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/terapia , Proteínas de Plantas/imunologia , Prunus persica/efeitos adversos , Imunoterapia Sublingual , Adulto , Antígenos de Plantas/administração & dosagem , Basófilos/imunologia , Basófilos/metabolismo , Feminino , Hipersensibilidade Alimentar/diagnóstico , Humanos , Imunoglobulina E/imunologia , Masculino , Hipersensibilidade a Amendoim/diagnóstico , Hipersensibilidade a Amendoim/imunologia , Hipersensibilidade a Amendoim/terapia , Proteínas de Plantas/administração & dosagem , Testes Cutâneos , Avaliação de Sintomas , Adulto Jovem
5.
Clin Exp Allergy ; 47(11): 1398-1408, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28618148

RESUMO

BACKGROUND: Recently, the nature of the lipid-ligand of Pru p 3, one of the most common plant food allergens in southern Europe, has been identified as a derivative of the alkaloid camptothecin bound to phytosphingosine. However, the origin of its immunological activity is still unknown. OBJECTIVE: We sought to evaluate the role of the Pru p 3 lipid-ligand in the immunogenic activity of Pru p 3. METHODS: In vitro cultures of different cell types (monocyte-derived dendritic cells [moDCs], PBMCs [peripheral blood mononuclear cells] and epithelial and iNKT-hybridoma cell lines) have been used to determine the immunological capacity of the ligand, by measuring cell proliferation, maturation markers and cytokine production. To study the capacity of the lipid-ligand to promote sensitization to Pru p 3 in vivo, a mouse model of anaphylaxis to peach has been produced and changes in the humoral and basophil responses have been analysed. RESULTS: The lipid-ligand of Pru p 3 induced maturation of moDCsc and proliferation of PBMCs. Its immunological activity resided in the phytosphingosine tail of the ligand. The adjuvant activity of the ligand was also confirmed in vivo, where the complex of Pru p 3-ligand induced higher levels of IgE than Pru p 3 alone. The immunological capacity of the Pru p 3 ligand was mediated by CD1d, as maturation of moDCs was inhibited by anti-CD1d antibodies and Pru p 3-ligand co-localized with CD1d on epithelial cells. Finally, Pru p 3-ligand presented by CD1d was able to interact with iNKTs. CONCLUSIONS AND CLINICAL RELEVANCE: The Pru p 3 lipid-ligand could act as an adjuvant to promote sensitization to Pru p 3, through its recognition by CD1d receptors. This intrinsic adjuvant activity of the accompanying lipid cargo could be a general essential feature of the mechanism underlying the phenomenon of allergenicity.


Assuntos
Antígenos de Plantas/imunologia , Hipersensibilidade Alimentar/imunologia , Proteínas de Plantas/imunologia , Sequência de Aminoácidos , Animais , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/metabolismo , Antígenos CD1d/imunologia , Antígenos CD1d/metabolismo , Antígenos de Plantas/administração & dosagem , Antígenos de Plantas/química , Citocinas/metabolismo , Europa (Continente) , Imunização , Imunoglobulina E/imunologia , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Ligantes , Lipídeos/imunologia , Ativação Linfocitária/imunologia , Camundongos , Modelos Moleculares , Células T Matadoras Naturais/imunologia , Células T Matadoras Naturais/metabolismo , Proteínas de Plantas/administração & dosagem , Proteínas de Plantas/química , Ligação Proteica , Conformação Proteica , Linfócitos T/imunologia , Linfócitos T/metabolismo
6.
Eur Ann Allergy Clin Immunol ; 49(1): 45-48, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28120607

RESUMO

More than 170 foods have been identified as being potentially allergenic. However, a minority of these foods cause the majority of reactions. Sweets are frequently implicated in allergic reactions in children with cow's milk, egg, nuts or fruits allergy, and they are the most relevant foods investigated as responsible allergens. We report an anaphylactic reaction to candies in an egg and peach allergic boy. We performed a study to identify responsible allergens for the reaction. We investigated hidden egg and peach allergens in the candies, but they were not found. Finally, the causative allergen resulted to be a vegetable protein from potato peel. We diagnosed a new allergy in our patient and Sol t 4 was identified as the responsible allergen of the anaphylactic reaction. We conclude that responsible allergens should always be studied and identified in whatever allergic reaction in order to prevent new reactions.


Assuntos
Alérgenos/imunologia , Anafilaxia/etiologia , Hipersensibilidade a Ovo/imunologia , Prunus persica/imunologia , Solanum tuberosum/imunologia , Humanos , Lactente , Masculino , Testes Cutâneos
7.
Clin Exp Allergy ; 46(8): 1111-9, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27017914

RESUMO

BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) act as cofactors worsening the allergic reactions induced by food allergens. AIM: The aim of this study was to evaluate the effect of both lysine acetylsalicylate (L-ASA) (non-selective cyclooxygenase (COX) inhibitor) and valdecoxib (selective COX-2 inhibitor) in basophils activated by peach lipid transfer protein (Pru p 3) in patients with food-dependent NSAID-induced anaphylaxis (FDNIA). METHODS: Twenty Pru p 3-allergic patients with FDNIA group, eleven peach anaphylaxis not exacerbated by NSAIDs (no-NSAID group) and 5 healthy volunteers were recruited. Basophil activation (BA) was measured as expression of CD63 (Flow(2) CAST(™) ; Bühlmann(®) ), after stimulation with Pru p 3, both alone and in combination with L-ASA (1.13, 3.38 and 6.78 mm) or valdecoxib (0.87, 7.8 and 31.25 µm). RESULTS: Basophils from no-NSAID group were significantly more reactive and sensitive to Pru p 3 than those from the FDNIA group. In both groups, an increase in BA was observed when basophils were exposed to Pru p 3 and L-ASA. In the FDNIA group, valdecoxib partially terminates the BA induced by Pru p 3, whereas in the no-NSAID group, a dual effect was observed depending on the concentration tested. CONCLUSIONS: This study indicates that subjects with food-induced anaphylaxis differ from FDNIA subjects in the higher reactivity and sensitivity of their basophils to allergen challenge. We have shown a direct effect of NSAIDs on basophils using a human model of FDNIA. Our results also suggest that selective COX2 inhibitors might be a safe alternative. BA test may be a useful tool in the study of the pathogenic mechanism of the cofactor phenomenon.


Assuntos
Anti-Inflamatórios não Esteroides/efeitos adversos , Basófilos/efeitos dos fármacos , Basófilos/imunologia , Hipersensibilidade Alimentar/imunologia , Imunoglobulina E/imunologia , Adulto , Alérgenos/imunologia , Anafilaxia/diagnóstico , Anafilaxia/imunologia , Biomarcadores , Progressão da Doença , Feminino , Alimentos/efeitos adversos , Hipersensibilidade Alimentar/diagnóstico , Humanos , Imunoglobulina E/sangue , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Adulto Jovem
8.
Artigo em Inglês | MEDLINE | ID: mdl-27012014

RESUMO

BACKGROUND: Component-based diagnosis on multiplex platforms is widely used in food allergy but its clinical performance has not been evaluated in nut allergy. OBJECTIVE: To assess the diagnostic performance of a commercial protein microarray in the determination of specific IgE (sIgE) in peanut, hazelnut, and walnut allergy. METHODS: sIgE was measured in 36 peanut-allergic, 36 hazelnut-allergic, and 44 walnut-allergic patients by ISAC 112, and subsequently, sIgE against available components was determined by ImmunoCAP in patients with negative ISAC results. ImmunoCAP was also used to measure sIgE to Ara h 9, Cora 8, and Jug r 3 in a subgroup of lipid transfer protein (LTP)-sensitized nut-allergic patients (positive skin prick test to LTP-enriched extract). sIgE levels by ImmunoCAP were compared with ISAC ranges. RESULTS: Most peanut-, hazelnut-, and walnut-allergic patients were sensitized to the corresponding nut LTP (Ara h 9, 66.7%; Cor a 8, 80.5%; Jug r 3, 84% respectively). However, ISAC did not detect sIgE in 33.3% of peanut-allergic patients, 13.9% of hazelnut-allergic patients, or 13.6% of walnut-allergic patients. sIgE determination by ImmunoCAP detected sensitization to Ara h 9, Cor a 8, and Jug r 3 in, respectively, 61.5% of peanut-allergic patients, 60% of hazelnut-allergic patients, and 88.3% of walnut-allergic patients with negative ISAC results. In the subgroup of peach LTP-sensitized patients, Ara h 9 sIgE was detected in more cases by ImmunoCAP than by ISAC (94.4% vs 72.2%, P < .05). Similar rates of Cora 8 and Jug r 3 sensitization were detected by both techniques. CONCLUSIONS: The diagnostic performance of ISAC was adequate for hazelnut and walnut allergy but not for peanut allergy. sIgE sensitivity against Ara h 9 in ISAC needs to be improved.


Assuntos
Alérgenos/imunologia , Corylus/imunologia , Juglans/imunologia , Hipersensibilidade a Noz/diagnóstico , Nozes/imunologia , Hipersensibilidade a Amendoim/diagnóstico , Proteínas de Plantas/imunologia , Análise Serial de Proteínas , Adulto , Biomarcadores/sangue , Estudos de Casos e Controles , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Testes Intradérmicos , Masculino , Região do Mediterrâneo , Pessoa de Meia-Idade , Hipersensibilidade a Noz/sangue , Hipersensibilidade a Noz/imunologia , Hipersensibilidade a Amendoim/sangue , Hipersensibilidade a Amendoim/imunologia , Valor Preditivo dos Testes , Espanha , Adulto Jovem
9.
Allergy ; 70(8): 1024-7, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25952012

RESUMO

Food allergy is recognized as a major public health issue, especially in early childhood. It has been hypothesized that early sensitization to food allergens maybe due to their ingestion as components dissolved in the milk during the breastfeeding, explaining reaction to a food, which has never been taken before. Thus, the aim of this work has been to detect the presence of the food allergens in breast milk by microarray technology. We produced a homemade microarray with antibodies produced against major food allergens. The antibody microarray was incubated with breast milk from 14 women collected from Fundación Jiménez Díaz Hospital. In this way, we demonstrated the presence of major foods allergens in breast milk. The analysis of allergens presented in breast milk could be a useful tool in allergy prevention and could provide us a key data on the role of this feeding in tolerance induction or sensitization in children.


Assuntos
Alérgenos/imunologia , Aleitamento Materno/efeitos adversos , Hipersensibilidade a Leite/diagnóstico , Leite Humano/imunologia , Fatores Etários , Alérgenos/análise , Criança , Pré-Escolar , Feminino , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/imunologia , Humanos , Incidência , Lactente , Masculino , Hipersensibilidade a Leite/epidemiologia , Hipersensibilidade a Leite/imunologia , Medição de Risco , Sensibilidade e Especificidade , Fatores Sexuais
10.
Pediatr Allergy Immunol ; 26(6): 497-502, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26046378

RESUMO

BACKGROUND: Peanut allergens are common triggers of food allergy. Analyses of sensitization patterns, relationships with other allergens, clinical symptoms, and variation with age are needed. We studied sensitization to Ara h 2, Ara h 9, and Pru p 3 in a peanut allergic children/adolescents and the relationship with peach and pollen. METHODS: Peanut allergic patients aged between 1 and 20 years old were classified into two groups: A) allergic to peanut only and B) allergic to peach and peanut. The IgE response was measured to Ara h 2, Ara h 9, and Pru p 3. RESULTS: Of 964 subjects evaluated, 28% were allergic to peanut. From this group, 68% were also sensitized to pollen. Urticaria was the most frequent entity followed by anaphylaxis and OAS. Fifty-eight percent had Ara h 2- and/or Ara h 9-specific IgE. More than half reported symptoms with peanut alone (Group A) and 35% to peanut and peach (Group B). We observed significant differences in sex, age, onset of symptoms, and sensitization to Artemisia between groups. IgE response to Ara h 2 was more frequent in Group A, and Ara h 9 and Pru p 3 in Group B. We observed a decrease in sensitization to Ara h 2 and an increase to Ara h 9 and Pru p 3 with increasing age. CONCLUSION: Peanut allergy is frequent in subjects with allergy to plant foods, with Ara h 2 and Ara h 9 being two important allergens. In younger patients, Ara h 2 predominates over Ara h 9. The reverse was observed in older patients.


Assuntos
Albuminas 2S de Plantas/imunologia , Antígenos de Plantas/imunologia , Glicoproteínas/imunologia , Imunoglobulina E/imunologia , Hipersensibilidade a Amendoim/imunologia , Proteínas de Plantas/imunologia , Adolescente , Fatores Etários , Anafilaxia/diagnóstico , Anafilaxia/imunologia , Biomarcadores/sangue , Criança , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Imunoglobulina E/sangue , Lactente , Testes Intradérmicos , Masculino , Hipersensibilidade a Amendoim/sangue , Hipersensibilidade a Amendoim/diagnóstico , Valor Preditivo dos Testes , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/imunologia , Testes Sorológicos , Espanha , Fatores de Tempo , Urticária/diagnóstico , Urticária/imunologia , Adulto Jovem
11.
Artigo em Inglês | MEDLINE | ID: mdl-26310039

RESUMO

Allergens come into contact with the immune system as components of a very diverse mixture. The most common sources are pollen grains, food, and waste. These sources contain a variety of immunomodulatory components that play a key role in the induction of allergic sensitization. The way allergen molecules bind to the cells of the immune system can determine the immune response. In order to better understand how allergic sensitization is triggered, we review the molecular mechanisms involved in the development of allergy and the role of immunomodulators in allergen recognition by innate cells.


Assuntos
Hipersensibilidade/imunologia , Imunidade Inata/fisiologia , Antígenos de Dermatophagoides/imunologia , Antígenos de Plantas/imunologia , Proteínas de Artrópodes/imunologia , Cisteína Endopeptidases/imunologia , Epigênese Genética , Humanos
12.
Artigo em Inglês | MEDLINE | ID: mdl-25898694

RESUMO

BACKGROUND AND OBJECTIVE: The allergenic potential of proteins can be altered under various physicochemical conditions. Glutathione (GSH) is a reducing agent that is used as an antioxidant in food products. We aimed to characterize the natural folding of peach proteins and test the allergenicity of reduced and natural Pru p 3, the major peach allergen. METHODS: Pru p 3 was purified from peach, and its conformation was analyzed by means of circular dichroism. Using a thiol fluorescent probe, reduced proteins were detected in fresh peach. GSH-reduced Pru p 3 was tested in vitro for T-cell proliferation and in vivo using skin prick testing. RESULTS: GSH-reduced Pru p 3 produced variable skin prick reactions in peach-allergic patients. The proliferative response of peripheral blood mononuclear cells from allergic patients to reduced Pru p 3 tended to be less intense, whereas secretion of the cytokines IFN-γ, IL-5, and IL-10 was comparable. In a pool of sera from peach-allergic patients, reduction hardly impaired IgE-binding. Moreover, the stability of reduced Pru p 3 to gastrointestinal digestion was similar to that of the natural form. CONCLUSIONS: GSH can at least transiently reduce Pru p 3. We found that the effect of reduction on the allergenicity of Pru p 3 varied. Therefore, as an additive, GSH does not seem to eliminate the risk of reactions for peach-allergic patients.


Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Proteínas de Transporte/imunologia , Glutationa/metabolismo , Proteínas de Plantas/imunologia , Prunus/imunologia , Adulto , Alérgenos/química , Alérgenos/metabolismo , Antígenos de Plantas/química , Antígenos de Plantas/metabolismo , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Proliferação de Células , Dicroísmo Circular , Citocinas/imunologia , Feminino , Hipersensibilidade Alimentar/imunologia , Humanos , Leucócitos Mononucleares/imunologia , Masculino , Pessoa de Meia-Idade , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Conformação Proteica , Prunus/metabolismo , Testes Cutâneos
13.
Artigo em Inglês | MEDLINE | ID: mdl-26727764

RESUMO

BACKGROUND: Baker's asthma (BA) is the most prevalent occupational respiratory disease in developed countries. It is caused by inhalation of wheat dust in the working environment and affects 1%-10% of workers in the baking industry. Diagnosis of BA is based on bronchial challenge with wheat, a technique that carries a high risk for patients. The wheat lipid transfer protein Tri a 14 is a major allergen in BA. OBJECTIVE: The aim of our study was to characterize Tri a 14 as a marker of BA in order to prevent patients from having to undergo bronchial challenge with wheat. METHODS: The study population comprised 55 patients selected at the Rio Hortega Hospital, Valladolid, Spain. Patients with BA were diagnosed using a skin prick test (SPT) with wheat and Tri a 14 and bronchial challenge test (BCT) with wheat. Patients with food allergy had a clear clinical history of allergy to peach confirmed by positive SPT to peach extract and Pru p 3. RESULTS: All patients in the BA group had a positive SPT result with wheat (100%), and most had positive results with Tri a 14 (95%). A positive BCT result with Tri a 14 was also observed in 22 of 27 of the patients with BA (82%). The response to Tri a 14 was specifically associated with BA. CONCLUSION: Tri a 14 is a good marker of BA and can be used in SPT and BCT as an alternative diagnostic method, thus avoiding bronchial challenge with wheat and reducing the risk associated with this technique.


Assuntos
Antígenos de Plantas/imunologia , Asma Ocupacional/diagnóstico , Proteínas de Transporte/imunologia , Hipersensibilidade a Trigo/diagnóstico , Adolescente , Adulto , Asma Ocupacional/imunologia , Testes de Provocação Brônquica , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Masculino , Pessoa de Meia-Idade , Testes Cutâneos , Hipersensibilidade a Trigo/imunologia , Adulto Jovem
14.
Allergy ; 69(7): 964-8, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24816395

RESUMO

Ara h 1, Ara h 2, and Ara h 3 are important sensitizers in peanut allergy. Ara h 9 has also been shown to be relevant in the Mediterranean area. We evaluated the basophil response to peanut allergens and Pru p 3 in Mediterranean patients: Group 1, peanut and peach allergy; Group 2, peanut allergy and tolerance to peach; Group 3, peach allergy and tolerance to peanut; Group 4, nonallergic subjects that tolerate both peanut and peach. Compared to controls (Group 4), there was an increased basophil activation with Ara h 2 (P = 0.031) and Pru p 3 (P = 0.009) in Group 1 and with Ara h 1 (P = 0.016), Ara h 2 (P = 0.001), and Ara h 9 (P = 0.016) in Group 2. Importantly, only Ara h 2 showed an increased activation (P = 0.009) in Group 2 compared to Group 3. Ara h 2 is the best discriminating allergen for peanut allergy diagnosis in a Mediterranean population showing two patterns: patients also allergic to peach, responding to Ara h 2 and Pru p 3, and patients allergic only to peanut, responding to Ara h 1, Ara h 2, and Ara h 9.


Assuntos
Albuminas 2S de Plantas/imunologia , Antígenos de Plantas/imunologia , Basófilos/imunologia , Glicoproteínas/imunologia , Hipersensibilidade a Amendoim/imunologia , Proteínas de Plantas/imunologia , Adulto , Alérgenos/imunologia , Teste de Degranulação de Basófilos , Reações Cruzadas/imunologia , Método Duplo-Cego , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Prunus/imunologia , Adulto Jovem
15.
Artigo em Inglês | MEDLINE | ID: mdl-25219106

RESUMO

BACKGROUND: IgE-mediated wheat allergy affects around 0.5% of the population, and current management is based on avoidance. We propose an active intervention to promote tolerance in wheat-allergic children. OBJECTIVES: To investigate the efficacy and safety of an oral immunotherapy (OIT) protocol with wheat to treat IgE-mediated wheat-allergic children. METHODS: Six wheat allergic patients assessed in a double-blind, placebo-controlled food challenge (DBPCFC) underwent wheat OIT with an up-dosing phase until 100 g of wheat was tolerated, followed by a 6-month maintenance phase. Tolerance to rye and oat was evaluated, as were specific IgE (sIgE) to wheat and other cereals and sIgE, slgG4, and sIgG1 to a panel of wheat proteins (alpha-amylase and trypsin inhibitors, wheat lipid transfer proteins, gliadins, and glutenins). RESULTS: Threshold doses in the wheat DBPCFC ranged from 6.6 g to 96.6 g. Five out of 6 (83%) patients successfully finished the up-dosing phase in 3 to 24 days; after a 6-month maintenance phase, all the patients maintained good tolerance of 100 g of wheat daily. Only 6.25% of doses in the up-dosing phase elicited mild adverse reactions. All 5 patients who successfully finished the up-dosing phase tolerated rye after OIT, and all but 1 tolerated oat as well. The median baseline wheat sIgE was 47.5 kU(A)/L, increasing to 84.55 kU(A)/L after up-dosing and decreasing to 28.75 kU(A)/L after 6 months of follow-up. None of the patients showed sIgE to 5-omega-gliadin, but alpha-amylase inhibitors were recognized by all patients. Specific IgG4 and sIgG1 increased in all patients. CONCLUSIONS: Our wheat OIT protocol was safe, efficient, and rapid. In our population, alpha-amylase was the major allergen.


Assuntos
Dessensibilização Imunológica/métodos , Imunoglobulina E/imunologia , Hipersensibilidade a Trigo/terapia , Administração Oral , Criança , Pré-Escolar , Método Duplo-Cego , Feminino , Humanos , Masculino , Hipersensibilidade a Trigo/imunologia
16.
Clin Exp Allergy ; 43(12): 1374-83, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24261947

RESUMO

BACKGROUND: Since intestinal absorption of food protein can trigger an allergic reaction, the effect of plant food allergen on intestinal epithelial cell permeability and its ability to cross the epithelial monolayer was evaluated. OBJECTIVE: To study the interaction of Pru p 3 with intestinal epithelium, its natural entrance, analyzing transport kinetics and cellular responses that trigger. METHODS: This was achieved using Pru p 3, the peach LTP, as a model. Enterocytic monolayers were established by culturing Caco 2 cells, as a model of enterocytes, on permeable supports that separate the apical and basal compartments. Pru p 3 was added to the apical compartment, the transepithelial resistance (TEER) was measured, and the transport was quantified. RESULTS: The peach allergen that crossed the cell monolayer was detected in the cell fraction and in the basal medium by immunodetection with specific antibodies and the quantity was measured by ELISA assay. Pru p 3 was able to cross the monolayer without disturbing the integrity of the tight junctions. This transport was significantly higher than that of a non-allergenic peach LTP, LTP1, and occurred via lipid raft pathway. The incubation of Caco 2 cells with Pru p 3 and LTP1 produced the expression of epithelial-specific cytokines TSLP, IL33 and IL25. CONCLUSION: These results suggest that Pru p 3 was able to cross the cell monolayer by the transcellular route and then induce the production of Th2 cytokines. The results of the present study represent a step towards clarifying the importance of Pru p 3 as a sensitizer. CLINICAL RELEVANCE: The capacity of food allergens to cross the intestinal monolayer could explain their high allergenic capacity and its fast diffusion through the body associating to severe symptoms.


Assuntos
Antígenos de Plantas/imunologia , Antígenos de Plantas/metabolismo , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/metabolismo , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Proteínas de Plantas/imunologia , Proteínas de Plantas/metabolismo , Transporte Biológico , Linhagem Celular , Citocinas/biossíntese , Hipersensibilidade Alimentar/genética , Humanos , Imunoglobulina E/imunologia , Imunoglobulina E/metabolismo , Cinética , Ligação Proteica/imunologia , Transporte Proteico , Proteólise , Células Th2/imunologia , Células Th2/metabolismo , Vesículas Transportadoras/metabolismo
17.
Clin Dev Immunol ; 2013: 385615, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24324505

RESUMO

Nowadays, treatment of food allergy only considered the avoidance of the specific food. However, the possibility of cross-reactivity makes this practice not very effective. Immunotherapy may exhibit as a good alternative to food allergy treatment. The use of hypoallergenic molecules with reduced IgE binding capacity but with ability to stimulate the immune system is a promising tool which could be developed for immunotherapy. In this study, three mutants of Pru p 3, the principal allergen of peach, were produced based on the described mimotope and T cell epitopes, by changing the specific residues to alanine, named as Pru p 3.01, Pru p 3.02, and Pru p 3.03. Pru p 3.01 showed very similar allergenic activity as the wild type by in vitro assays. However, Pru p 3.02 and Pru p 3.03 presented reduced IgE binding with respect to the native form, by in vitro, ex vivo, and in vivo assays. In addition, Pru p 3.03 had affected the IgG4 binding capacity and presented a random circular dichroism, which was reflected in the nonrecognition by specific antibodies anti-Pru p 3. Nevertheless, both Pru p 3.02 and Pru p 3.03 maintained the binding to IgG1 and their ability to activate T lymphocytes. Thus, Pru p 3.02 and Pru p 3.03 could be good candidates for potential immunotherapy in peach-allergic patients.


Assuntos
Antígenos de Plantas/imunologia , Proteínas Mutantes/imunologia , Proteínas de Plantas/imunologia , Vacinas/imunologia , Adolescente , Adulto , Sequência de Aminoácidos , Anticorpos Bloqueadores/imunologia , Especificidade de Anticorpos/imunologia , Antígenos de Plantas/química , Antígenos de Plantas/genética , Feminino , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/terapia , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoterapia , Ativação Linfocitária/imunologia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Proteínas de Plantas/química , Proteínas de Plantas/genética , Ligação Proteica/imunologia , Linfócitos T/imunologia , Adulto Jovem
18.
Int Arch Allergy Immunol ; 159(4): 346-54, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22832505

RESUMO

BACKGROUND: Patients with pollinosis develop symptoms after intake of plant food more often than the general population. In order to study the prevalence of the presentation of allergic symptoms to plant foods in pollinosis, we selected a representative sample of the population from our Mediterranean area. METHODS: All patients completed a questionnaire, provided a blood sample and underwent a battery of skin and other complementary tests (prick-prick, oral challenge test) when necessary. The pollen counts were obtained from the Elche pollen station. In addition, sera from a subgroup of patients were checked with an allergen molecule panel on an Advia Centaur XP platform. RESULTS: Of the final sample (n = 233), 39.9% of the patients with pollinosis were sensitized and 30.9% had clinical allergy to at least one of the plant foods studied. Regression analysis showed that age and sensitization to the extracts of Platanus acerifolia and Artemisia vulgaris were the most important variables for discriminating between groups. Patients with pollinosis at a risk of allergy to plant foods had significantly higher Pru p 3 values [odds ratio (OR) 3.3; 95% confidence interval (95% CI) 2.3-4.8], and the value increased according to the number of plant food sensitizations. CONCLUSION: Plant food allergy is more common in patients with pollinosis than in the general population. The use of the London plain tree (P. acerifolia) and mugwort (A. vulgaris) in the skin tests may help identify such patients in our Mediterranean area, but determination of rPru p 3 could also be very useful in patients suspected of having plant food allergy.


Assuntos
Artemisia/imunologia , Hipersensibilidade Alimentar/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Árvores/imunologia , População Branca , Adolescente , Adulto , Idoso , Alérgenos/imunologia , Artemisia/química , Comorbidade , Feminino , Hipersensibilidade Alimentar/complicações , Hipersensibilidade Alimentar/etnologia , Humanos , Masculino , Região do Mediterrâneo/epidemiologia , Pessoa de Meia-Idade , Prevalência , Análise de Regressão , Rinite Alérgica Sazonal/complicações , Rinite Alérgica Sazonal/etnologia , Testes Cutâneos , Árvores/química
20.
J Investig Allergol Clin Immunol ; 21(2): 81-92; quiz 94, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21462797

RESUMO

Baker's asthma is a frequent occupational allergic disease caused mainly by inhalation of cereal flour, particularly wheat flour. This review deals with the current diagnosis and immunomodulatory treatments, as well as the role of wheat allergens as molecular tools to enhance management and knowledge of this disease. The review also discusses the current status of several salt-soluble proteins (albumins and globulins)--cereal alpha-amylase/trypsin inhibitors, peroxidase, thioredoxin, nonspecific lipid transfer protein, serine proteinase inhibitor, and thaumatin-like protein-as well as salt-insoluble storage proteins (prolamins, namely, gliadins and glutenins) as allergens associated with baker's asthma. Finally, current limitations to using these proteins as molecular tools for diagnosis and immunotherapy are highlighted.


Assuntos
Alérgenos/imunologia , Asma/diagnóstico , Asma/imunologia , Doenças Profissionais/imunologia , Triticum/imunologia , Asma/terapia , Humanos , Imunoterapia , Triticum/química , Hipersensibilidade a Trigo/imunologia , Hipersensibilidade a Trigo/terapia
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