RESUMO
The protein content and allergen composition was studied of cashews from 8 different origins (Benin, Brazil, Ghana, India, Ivory Coast, Mozambique, Tanzania, Vietnam), subjected to different in-shell heat treatments (steamed, fried, drum-roasted). On 2D electrophoresis, 9 isoforms of Ana o 1, 29 isoforms of Ana o 2 (11 of the acidic subunit, 18 of the basic subunit), and 8 isoforms of the large subunit of Ana o 3 were tentatively identified. Based on 1D and 2D electrophoresis, no difference in allergen content (Ana o 1, 2, 3) was detected between the cashews of different origins (P > 0.5), some small but significant differences were detected in allergen solubility between differently heated cashews. No major differences in N- and C-terminal microheterogeneity of Ana o 3 were detected between cashews of different origins. Between the different heat treatments, no difference was detected in glycation, pepsin digestibility, or IgE binding of the cashew proteins.
Assuntos
Alérgenos/imunologia , Anacardium/química , Manipulação de Alimentos , Nozes/química , Anacardium/imunologia , Antígenos de Plantas/imunologia , Benin , Brasil , Côte d'Ivoire , Gana , Humanos , Índia , Moçambique , Hipersensibilidade a Noz/imunologia , Nozes/imunologia , Proteínas de Plantas/imunologia , Tanzânia , VietnãRESUMO
Cashew nut allergy is the second most commonly reported tree nut allergy. Traditional allergen immunotherapy presents several clinical drawbacks that can be reduced by using nanoparticles-basedallergen-delivery systems, modulating the immune response towards a protective one. In this context, the goal of this work was to assess the potential of poly(anhydride) nanoparticles (NP) for cashew nut oral immunization. Cashew nut allergens-loaded nanoparticles (CNE-NP) were prepared by solvent displacement method. After nanoparticles characterization, oral immunomodulation ability was evaluated in BALB/c mice. Our results demonstrated that CNE-NP induced a higher Th1/Th2 ratio in comparison with animals immunized with free cashew nut proteins. Indeed, a decrease in splenic Th2 cytokines (IL-4, IL-5, and IL-13), and an enhancement of pro-Th1 (IL-12 and IFN-γ) and regulatory (IL-10) cytokines was observed. Furthermore, mice orally immunized with CNE-NP presented an increased expansion of CD4+ T regulatory cells, such as CD4+Foxp3+ and CD4+LAP+, in the mesenteric lymph nodes. In conclusion, oral immunization with a single dose of poly(anhydride) nanoparticles loaded with cashew nut proteins leaded to a pro-Th1 and Treg immune response. Furthermore, their immunomodulatory properties could be introduced as a new approach for management of cashew nut allergy.
Assuntos
Anacardium/imunologia , Anidridos/imunologia , Nanopartículas/efeitos adversos , Hipersensibilidade a Noz/imunologia , Nozes/imunologia , Linfócitos T Reguladores/imunologia , Células Th1/imunologia , Administração Oral , Alérgenos/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Citocinas/imunologia , Dessensibilização Imunológica/métodos , Linfonodos/imunologia , Camundongos , Camundongos Endogâmicos BALB CAssuntos
Albuminas 2S de Plantas/imunologia , Antígenos de Plantas/imunologia , Imunoglobulina E/imunologia , Hipersensibilidade a Noz/diagnóstico , Hipersensibilidade a Noz/imunologia , Proteínas Recombinantes , Humanos , Imunoglobulina E/sangue , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Lipids are required for mice sensitization to Ber e 1, Brazil nut major allergen. Here, we characterized different lipid fractions extracted from Brazil nuts and the lipid-binding ability of Ber e 1. Further, we determined their in vivo ability to induce Ber-specific anaphylactic antibodies and the role of invariant natural killer T (iNKT) cells in this process. METHODS: Wild-type (WT) and iNKT cell-deficient mice were sensitized with Ber e 1 and specific lipid fractions, and anaphylactic antibodies were measured by enzyme-linked immunosorbent assay (ELISA) and passive cutaneous anaphylaxis (PCA). The lipid-binding characteristic of Ber e 1 (Ber) was established by using fluorescent probes and (15) N-labeled NMR. In vitro production of IL-4 was determined in Ber/lipid C-stimulated mouse iNKT cells and human T-cell lines containing NKTs primed with CD1d+C1R transfectants by flow cytometry and ELISA, respectively. RESULTS: Only one specific lipid fraction (lipid C), containing neutral and common phospholipids, induced Ber anaphylactic antibodies in mice. Ber e 1 has a lipid-binding site, and our results indicated an interaction between Ber e 1 and lipid C. iNKT-deficient mice produced lower levels of anaphylactic antibodies than WT mice. In vitro, Ber/lipid C-stimulated murine iNKT cells produced IL-4 but not IFN-gamma. Human T-cell lines derived from nut-allergic patients produced IL-4 to Ber/lipid C in a CD1d- and dose-dependent manner. CONCLUSION: Lipid fraction C from Brazil nut presents an essential adjuvant activity to Ber e 1 sensitization, and iNKT cells play a critical role in the development of Brazil nut-allergic response.
Assuntos
Lipídeos/imunologia , Células T Matadoras Naturais/imunologia , Hipersensibilidade a Noz/imunologia , Albuminas 2S de Plantas/química , Albuminas 2S de Plantas/imunologia , Adolescente , Adulto , Alérgenos/imunologia , Animais , Anticorpos/sangue , Anticorpos/imunologia , Antígenos de Plantas/química , Antígenos de Plantas/imunologia , Sítios de Ligação , Feminino , Humanos , Interações Hidrofóbicas e Hidrofílicas , Ativação Linfocitária/imunologia , Masculino , Camundongos , Hipersensibilidade a Noz/metabolismo , Ligação Proteica , Células Th2/imunologia , Adulto JovemRESUMO
BACKGROUND: Lipids, particularly bacterial lipopolysaccharide, can impact on immune responses to proteins, with low doses enhancing type 2 responses. OBJECTIVE: We have examined the influence of natural plant lipid extracts on antibody responses provoked in mice by recombinant Ber e 1, the major allergen in Brazil nuts. METHODS: BALB/c strain mice were immunized (by intraperitoneal injection) with natural or recombinant Ber e l produced in Pichia pastoris and admixed with various lipid fractions isolated from Brazil nuts. Serum samples were analysed for specific IgE antibody by homologous passive cutaneous anaphylaxis assay and for IgG by enzyme-linked immunosorbant assay. RESULTS: Exposure to recombinant (lipid-free) Ber e 1 alone failed to induce detectable IgG or IgE antibody. Co-administration of the total lipid fraction (with reduced triglyceride levels), sterol-rich, or polar lipid fractions, resulted in marked adjuvant effects on IgG and IgE. However, the beta-sitosterol and glycolipid-rich fractions were associated with only low-level IgG antibody, and had little impact on IgE antibody production. Natural Ber e 1 containing endogenous lipids also provoked IgG and IgE antibody responses. Identical IgE and IgG antibody responses were detected regardless of whether natural or recombinant Ber e 1 was used as substrates for analyses. CONCLUSION: Endogenous Brazil nut lipids are required for the induction of optimal antibody responses to Ber e 1 in the BALB/c strain mouse. Appropriate antibody binding sites are present on both natural and recombinant forms of Ber e 1, suggesting that the impact of lipid is at the induction phase, rather than antibody recognition, and is possibly required for efficient antigen presentation.
Assuntos
Albuminas/imunologia , Alérgenos/imunologia , Lipídeos/imunologia , Hipersensibilidade a Noz/imunologia , Precursores de Proteínas/imunologia , Albuminas 2S de Plantas , Adjuvantes Imunológicos , Animais , Antígenos de Plantas , Bertholletia/imunologia , Feminino , Imunoglobulina E/biossíntese , Imunoglobulina G/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Plantas/imunologia , Proteínas Recombinantes/imunologiaRESUMO
The diagnosis and management of nut allergy can be difficult because of the possible severity of the clinical manifestations and the cross reactivity between different species. We report a case of anaphylaxis due to skin testing in a young adult with clinically ascertained walnut allergy. After an episode of anaphylaxis due to walnut ingestion, a routine diagnostic workup was carried out, involving skin prick test with commercial extracts, prick by prick with fresh food and CAP-RAST assay for different nuts. Immediately after pricking with fresh Brazil nut, a severe episode of anaphylaxis occurred, that required epinephrine and intravenous steroids. The subject had never eaten Brazil nut before. Therefore we hypothesize a cross reactivity effect, since this phenomenon is well known for tree nuts. Our case suggests that in vivo diagnosis, especially if fresh nuts are used, should be performed only if adequate equipment to treat anaphylaxis is available.