RESUMEN
BACKGROUND: Ragweed (Ambrosia elatior) has become invasive in Europe, causing significant respiratory issues. Subcutaneous allergen immunotherapy (SCIT) has long been used to manage pollen allergies, but sublingual immunotherapy (SLIT) has gained interest. OBJECTIVE: This study aimed to evaluate the clinical benefits of ragweed SLIT under real-world in a cohort of Hungarian patients allergic to ragweed pollen. METHODS: We retrospectively reviewed the clinical records of 57 patients during the 2015 and 2016 ragweed pollen seasons. Patients were divided into two groups: Group 1 (n = 29), who had not received immunotherapy, and Group 2 (n = 28), who had previously undergone immunotherapy with another sublingual preparation. All patients were treated with Oraltek® ragweed for 4-6 months, initiating 2-4 months before the pollen season and rest of the period was 2 months of the 2016 pollen season. Symptom score (SS), medication score (MS), and combined symptom and medication score (CSMS) were evaluated intra- and intergroup. RESULTS: Pollen counts were consistent between 2015 and 2016. All patients showed significant improvement in SS, MS, and CSMS, with a large effect size (>0.8). Group 2 had significantly lower SS and CSMS in 2015 because of prior immunotherapy. By 2016, both groups exhibited marked improvements, with Group 1 showing a 75% improvement in CSMS. No local or systemic reactions were recorded, indicating a high safety profile. CONCLUSIONS: Ragweed SLIT significantly improved symptoms and reduced use of medication in patients allergic to ragweed pollen. The treatment was effective even in patients with previous immunotherapy, with a high benefit-risk ratio demonstrated by the absence of adverse reactions. These findings support the use of Oraltek SLIT for managing ragweed pollen allergy.
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Alérgenos , Ambrosia , Antígenos de Plantas , Rinitis Alérgica Estacional , Inmunoterapia Sublingual , Humanos , Inmunoterapia Sublingual/métodos , Masculino , Femenino , Estudios Retrospectivos , Rinitis Alérgica Estacional/terapia , Rinitis Alérgica Estacional/inmunología , Adulto , Ambrosia/inmunología , Alérgenos/inmunología , Alérgenos/administración & dosificación , Hungría , Antígenos de Plantas/inmunología , Antígenos de Plantas/administración & dosificación , Persona de Mediana Edad , Adulto Joven , Extractos Vegetales/administración & dosificación , Resultado del Tratamiento , Adolescente , Polen/inmunologíaAsunto(s)
Anafilaxia , Citrus sinensis , Hipersensibilidad a los Alimentos , Giberelinas , Humanos , Anafilaxia/diagnóstico , Anafilaxia/etiología , Anafilaxia/inmunología , Hipersensibilidad a los Alimentos/inmunología , Hipersensibilidad a los Alimentos/diagnóstico , Citrus sinensis/inmunología , Citrus sinensis/efectos adversos , Giberelinas/efectos adversos , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangre , Proteínas de Plantas/inmunología , Proteínas de Plantas/efectos adversos , Femenino , Masculino , Alérgenos/inmunología , Alérgenos/efectos adversos , Antígenos de Plantas/inmunología , Pruebas CutáneasRESUMEN
BACKGROUND: Cross-reactivity between nonspecific lipid transfer proteins could cause anaphylaxis, further influencing food avoidance and nutrient deficiencies. The one affecting olive pollen (Ole e 7) and peach (Pru p 3) may underlie a variety of pollen-food syndromes, though a deep molecular analysis is necessary. METHODS: Three Ole e 7-monosensitised patients (MON_OLE), three Pru p 3-monosensitised patients (MON_PRU) and three bisensitised patients (BI) were selected. For epitope mapping, both digested proteins were incubated with patient sera, and the captured IgE-bound peptides were characterised by LC-MS. RESULTS: The analysis revealed two Ole e 7 epitopes and the three Pru p 3 epitopes previously described. Interestingly, the "KSALALVGNKV" Ole e 7 peptide was recognised by MON_OLE, BI and MON_PRU patients. Conversely, all patients recognised the "ISASTNCATVK" Pru p 3 peptide. Although complete sequence alignment between both proteins revealed 32.6% identity, local alignment considering seven residue fragments showed 50 and 57% identity when comparing "ISASTNCATVK" with Ole e 7 and "KSALALVGNKV" with Pru p 3. CONCLUSIONS: This study mapped sIgE-Ole e 7-binding epitopes, paving the way for more precise diagnostic tools. Assuming non-significant sequence similarity, structural homology and shared key residues may underlie the potential cross-reactivity between Ole e 7 and Pru p 3 nsLTPs.
Asunto(s)
Antígenos de Plantas , Reacciones Cruzadas , Hipersensibilidad a los Alimentos , Inmunoglobulina E , Olea , Proteínas de Plantas , Polen , Prunus persica , Humanos , Antígenos de Plantas/inmunología , Hipersensibilidad a los Alimentos/inmunología , Polen/inmunología , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangre , Olea/inmunología , Proteínas de Plantas/inmunología , Femenino , Masculino , Prunus persica/inmunología , Mapeo Epitopo , Adulto , Rinitis Alérgica Estacional/inmunología , Secuencia de Aminoácidos , Epítopos/inmunología , Alérgenos/inmunología , Persona de Mediana Edad , Proteínas Portadoras/inmunologíaRESUMEN
The topic of ragweed pollen (RW) versus house dust mites (HDMs) has often been deliberated, but the increasing incidence of co-sensitization between them has been scarcely addressed. Utilizing Sprague Dawley rats, we explored the effects of co-sensitization with the combination of HDMs and RW pollen extracts in correlation with high-fructose diet (HFrD) by in vitro tracheal reactivity analysis in isolated organ bath and biological explorations. Our findings unveiled interrelated connections between allergic asthma, dyslipidemia, and HFrD-induced obesity, shedding light on their compounding role through inflammation. The increased CRP values and airway hyperresponsiveness to the methacholine challenge suggest a synergistic effect of obesity on amplifying the existing inflammation induced by asthma. One of the major outcomes is that the co-sensitization to HDMs and RW pollen led to the development of a severe allergic asthma phenotype in rats, especially in those with HFrD. Therefore, the co-sensitization to these allergens as well as the HFrD may play a crucial role in the modulation of systemic inflammation, obesity, and airway reactivity.
Asunto(s)
Biomarcadores , Fructosa , Pyroglyphidae , Ratas Sprague-Dawley , Animales , Ratas , Biomarcadores/sangre , Pyroglyphidae/inmunología , Asma/inmunología , Asma/sangre , Asma/etiología , Masculino , Alérgenos/inmunología , Obesidad/inmunología , Obesidad/sangre , Antígenos de Plantas/inmunología , Polen/inmunología , Extractos VegetalesRESUMEN
IgE-mediated wheat allergy can take on various forms, including childhood food allergy to wheat, wheat-dependent exercise-induced anaphylaxis in young adults, baker's respiratory allergy/asthma in workers exposed to wheat flour inhalation, and contact urticaria that is caused by hydrolyzed wheat proteins in some cosmetics, and that is sometimes associated with a food allergy. Singleplex and multiplex immunoassays detect specific IgE antibodies to wheat allergenic molecular biomarkers such as omega-5 gliadin Tri a 19, lipid transfer protein Tri a 14, and alpha-amylase inhibitors. The fluorescence enzyme immunoassay with capsulated cellulose polymer solid-phase coupled allergens is a commonly used singleplex assay. Multiplex methods include the ELISA-based macroarray immunoassay using nano-bead technology and a microarray immunoassay on polymer-coated slides. Another promising diagnostic tool is the basophil activation test performed with omega-5 gliadin and other wheat protein types. Detailed comprehension of the structural and immunological features of the numerous wheat allergens significant in clinical settings is imperative for advancing diagnostic biomarkers for IgE-mediated wheat allergies.
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Alérgenos , Biomarcadores , Gliadina , Inmunoglobulina E , Hipersensibilidad al Trigo , Hipersensibilidad al Trigo/diagnóstico , Hipersensibilidad al Trigo/inmunología , Humanos , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangre , Alérgenos/inmunología , Gliadina/inmunología , Triticum/inmunología , Antígenos de Plantas/inmunología , Inmunoensayo/métodosRESUMEN
Peanut allergy has garnered worldwide attention due to its high incidence rate and severe symptoms, stimulating the demand for the ultrasensitive detection method of peanut allergen. Herein, we successfully developed a novel electrochemical aptasensor for ultrasensitive detection Ara h1, a major allergenic protein present in peanuts. A conductive nickel atoms Anchored Hydrogen-Bonded Organic Frameworks (PFC-73-Ni) were utilized as excellent electrocatalysts toward hydroquinone (HQ) oxidation to generate a readable current signal. The developed electrochemical aptasensor offers wide linear range (1-120 nM) and low detection limit (0.26 nM) for Ara h1. This method demonstrated a recovery rate ranging from 95.00% to 107.42% in standard addition detection of non-peanut food samples. Additionally, the developed electrochemical method was validated with actual samples and demonstrated good consistency with the results obtained from a commercial ELISA kit. This indicates that the established Ara h1 detection method is a promising tool for peanut allergy prevention.
Asunto(s)
Antígenos de Plantas , Arachis , Técnicas Electroquímicas , Antígenos de Plantas/análisis , Antígenos de Plantas/inmunología , Antígenos de Plantas/química , Arachis/química , Arachis/inmunología , Enlace de Hidrógeno , Glicoproteínas/química , Glicoproteínas/análisis , Límite de Detección , Estructuras Metalorgánicas/química , Proteínas de Plantas/química , Proteínas de Plantas/inmunología , Proteínas de Plantas/análisis , Técnicas Biosensibles/instrumentación , Alérgenos/análisis , Alérgenos/química , Alérgenos/inmunología , Porosidad , Aptámeros de Nucleótidos/química , Proteínas de la MembranaRESUMEN
Due to the benefits of tomato as an antioxidant and vitamin source, allergy to this vegetable food is a clinically concerning problem. Sola l 7, a class I lipid transfer protein found in tomato seeds, has been identified as an allergen linked to severe anaphylaxis. However, the role of lipid binding in Sola l 7-induced allergy remains unclear. Here, the three-dimensional structure of recombinant Sola l 7 (rSola l 7) has been elucidated using nuclear magnetic resonance spectroscopy (NMR). Its interaction with free fatty acids has been deeply studied; fluorescence emission spectroscopy revealed that different long-chain fatty acids interact with the protein, affecting the only tyrosine residue present in Sola l 7. On the contrary, no changes in the overall secondary structure were observed after the analysis of the circular dichroism spectra in the presence of fatty acids. Unsaturated oleic and linoleic fatty acids presented higher affinity and promoted more significant changes than saturated or short-chain fatty acids. 1H-15N HSQC NMR spectra allowed to determine the regions of the protein that were modified when rSola l 7 interacts with the fatty acids, suggesting epitope modification after the interaction. For corroboration, IgG and IgE binding to rSola l 7 were assessed in the presence of free fatty acids, revealing that both IgE and IgG binding were significantly lower than in their absence, suggesting a potential protective role of unsaturated fatty acids in tomato allergy.
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Proteínas Portadoras , Hipersensibilidad a los Alimentos , Proteínas de Plantas , Semillas , Solanum lycopersicum , Solanum lycopersicum/química , Hipersensibilidad a los Alimentos/inmunología , Proteínas de Plantas/química , Proteínas de Plantas/inmunología , Proteínas Portadoras/química , Humanos , Semillas/química , Inmunoglobulina E/inmunología , Inmunoglobulina E/química , Inmunoglobulina E/metabolismo , Ácidos Grasos/química , Antígenos de Plantas/química , Antígenos de Plantas/inmunología , Alérgenos/química , Alérgenos/inmunología , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Inmunoglobulina G/química , Resonancia Magnética Nuclear BiomolecularRESUMEN
PURPOSE OF REVIEW: Plant-derived foods are one of the most common causative sources of food allergy in China, with a significant relationship to pollinosis. This review aims to provide a comprehensive overview of this food-pollen allergy syndrome and its molecular allergen diagnosis to better understand the cross-reactive basis. RECENT FINDINGS: Food-pollen cross-reactivity has been mainly reported in Northern China, Artemisia pollen is the major related inhalant source, followed by tree pollen (Betula), while grass pollen plays a minor role. Pollen allergy is relatively low in Southern China, with allergies to grass pollen being more important than weed and tree pollens. Rosaceae fruits and legume seeds stand out as major related allergenic foods. Non-specific lipid transfer protein (nsLTP) has been found to be the most clinically relevant cross-reacting allergenic component, able to induce severe reactions. PR-10, profilin, defensin, chitinase, and gibberellin-regulated proteins are other important cross-reactive allergen molecules. Artemisia pollen can induce allergenic cross-reactions with a wide range of plant-derived foods in China, and spring tree pollens (Betula) are also important. nsLTP found in both pollen and plant-derived food is considered the most significant allergen in food pollen cross-reactivity. Component-resolved diagnosis with potential allergenic proteins is recommended to improve diagnostic accuracy and predict the potential risk of causing allergic symptoms.
Asunto(s)
Alérgenos , Reacciones Cruzadas , Hipersensibilidad a los Alimentos , Polen , Humanos , Reacciones Cruzadas/inmunología , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/inmunología , China , Alérgenos/inmunología , Polen/inmunología , Rinitis Alérgica Estacional/inmunología , Rinitis Alérgica Estacional/diagnóstico , Antígenos de Plantas/inmunología , Artemisia/inmunología , Proteínas de Plantas/inmunologíaRESUMEN
This study investigated the effects of genetic diversity in the allergenicity of peanut and assessed the allergenic capacity of six Arachis hypogaea accessions using a Balb/c mouse model. It also explored potential cross-reactivities between Ara h 3 (peanut allergen) and Gly m (soybean allergen) using computational tools. Female Balb/c mice were injected with peanut protein extracts and alum. Serum-specific antibodies (IgE, IgGt, IgG1, IgG2a) were measured using ELISA, and allergic protein profiles were examined via western blot. Structural homology, B cell epitopes, and molecular interactions between Ara h 3 and Gly m with human IgE were also investigated. The mice developed high sIgE and sIgG1 responses, with antibodies recognizing 19 bands on western blot. Notably, Saharan accessions showed unique features such as no bands on western blot profiles, reduced anaphylactic symptoms, lower IgE titers, and less intestinal tissue damage. Molecular docking results suggest significant cross-allergenicity, supported by allergenicity predictions and structural homology analysis. This comprehensive analysis provides insights into shared epitopes, potential competition for binding sites, and molecular dynamics of cross-reactive responses, enhancing understanding of food allergen interactions. The study recommends using Algerian Sahara peanut accessions in breeding, genomics studies, and industry for safer peanut options for individuals with allergies. SIGNIFICANCE: The significance of this study lies in its contribution to addressing a major public health issue: peanut allergy, which represents a significant cause of anaphylaxis affecting numerous individuals and families worldwide. By exploring the genetic diversity of peanut proteins and identifying hypoallergenic accessions through experimental and computational approaches, this research offers valuable insights for mitigating allergic reactions. The findings highlight that certain accessions from the Saharan region exhibit reduced allergenicity, resulting in attenuated anaphylactic symptoms, lower IgE levels, and reduced intestinal damage in murine models. Furthermore, the study's in silico analysis sheds light on the issue of cross-reactivity between peanut and soybean allergens, providing crucial information for understanding allergen interactions at the molecular level. Overall, this research contributes to advancing knowledge in the field of food allergen research and has practical implications for improving the quality of life for individuals allergic to peanuts, particularly through the selection of safer peanut varieties and their cultivation.
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Antígenos de Plantas , Arachis , Variación Genética , Hipersensibilidad al Cacahuete , Proteínas de Plantas , Animales , Femenino , Humanos , Ratones , Alérgenos/inmunología , Antígenos de Plantas/inmunología , Antígenos de Plantas/genética , Arachis/inmunología , Arachis/genética , Simulación por Computador , Reacciones Cruzadas/inmunología , Modelos Animales de Enfermedad , Glicoproteínas/inmunología , Glicoproteínas/genética , Inmunoglobulina E/inmunología , Ratones Endogámicos BALB C , Simulación del Acoplamiento Molecular , Hipersensibilidad al Cacahuete/inmunología , Proteínas de Plantas/inmunología , Proteínas de Plantas/genéticaRESUMEN
PURPOSE OF REVIEW: To provide an update on the diagnosis of non-specific Lipid Transfer Protein (nsLTP) allergy. RECENT FINDINGS: More publications report the presence of nsLTP allergy in Northern European countries and nsLTP sensitisation in children. Individuals are more likely to have severe reactions if there is recognition of increasing numbers of LTP components. Diagnosis is problematic; not all those with nsLTP allergy will have a positive test to a peach extract containing Pru p 3, the peach nsLTP. Sensitisation to nsLTP is being reported in more countries, including to the nsLTP in Cannabis Sativa in North America. Meals containing multiple nsLTP foods are more likely to be involved in co-factor reactions. Component-resolved diagnostics are superior to skin prick tests, to determine sensitisation to the individual nsLTP allergens causing symptoms and, in the future, the Basophil Activation test may best discriminate between sensitization and clinical allergy.
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Antígenos de Plantas , Proteínas Portadoras , Hipersensibilidad a los Alimentos , Pruebas Cutáneas , Humanos , Proteínas Portadoras/inmunología , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/inmunología , Antígenos de Plantas/inmunología , Alérgenos/inmunología , Proteínas de Plantas/inmunología , Niño , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangreRESUMEN
BACKGROUND: Allergy to peanuts and tree nuts is a common cause of food allergy in Spain, with lipid transfer proteins (LTP) being the most frequently recognized panallergen. LTP sensitization often leads to multiple food group sensitivities, resulting in overly restrictive diets that hinder patient's quality of life. This study aimed to assess the tolerance of peanuts and tree nuts (hazelnuts and walnuts) in children sensitized to LTP, potentially mitigating the need for such diets. METHODS: This prospective study enrolled individuals diagnosed with allergy to peanuts, hazelnuts, or walnuts. Data were collected from medical records, including demographics and clinical history. Allergological assessment comprised skin prick tests using commercial extracts and the nuts in question, alongside measurements of total and specific IgE to nuts and their primary molecular components. Participants showing positive LTP sensitization without sensitization to seed storage proteins underwent open oral nut challenges. RESULTS: A total of 75 individuals labeled as allergic to peanuts, 44 to hazelnuts, and 51 to walnuts were included. All of them underwent an open oral provocation test with the incriminated nut, showing a high tolerance rate. Peanut was tolerated by 98.6% of patients, 97.72% tolerated hazelnut, and 84.3% tolerated walnut. CONCLUSION: The findings suggest that the majority of patients allergic to peanuts, hazelnuts, or walnuts, due to LTP sensitization and lacking IgE reactivity to seed storage proteins, can tolerate these nuts. This supports the need for personalized nut tolerance assessments to avoid unnecessary dietary restrictions.
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Arachis , Proteínas Portadoras , Tolerancia Inmunológica , Inmunoglobulina E , Hipersensibilidad a la Nuez , Pruebas Cutáneas , Humanos , Masculino , Femenino , Proteínas Portadoras/inmunología , Niño , España , Estudios Prospectivos , Preescolar , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Hipersensibilidad a la Nuez/inmunología , Hipersensibilidad a la Nuez/diagnóstico , Arachis/inmunología , Hipersensibilidad al Cacahuete/inmunología , Hipersensibilidad al Cacahuete/diagnóstico , Alérgenos/inmunología , Juglans/inmunología , Nueces/inmunología , Adolescente , Corylus/inmunología , Hipersensibilidad a Nueces y Cacahuetes/inmunología , Antígenos de Plantas/inmunologíaRESUMEN
(1) Peanut allergy is associated with high risk of anaphylaxis which could be prevented by oral immunotherapy. Patients eligible for immunotherapy are selected on the basis of a food challenge, although currently the assessment of antibodies against main peanut molecules (Ara h 1, 2, 3 and 6) is thought to be another option. (2) The current study assessed the relationship between the mentioned antibodies, challenge outcomes, skin tests and some other parameters in peanut-sensitized children. It involved 74 children, divided into two groups, based on their response to a food challenge. (3) Both groups differed in results of skin tests, levels of component-specific antibodies and peanut exposure history. The antibody levels were then used to calculate thresholds for prediction of challenge results or symptom severity. While the antibody-based challenge prediction revealed statistical significance, it failed in cases of severe symptoms. Furthermore, no significant correlation was observed between antibody levels, symptom-eliciting doses and the risk of severe anaphylaxis. Although in some patients it could result from interference with IgG4, the latter would not be a universal explanation of this phenomenon. (4) Despite some limitations, antibody-based screening may be an alternative to the food challenge, although its clinical relevance still requires further studies.
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Arachis , Hipersensibilidad al Cacahuete , Humanos , Hipersensibilidad al Cacahuete/diagnóstico , Hipersensibilidad al Cacahuete/inmunología , Niño , Femenino , Masculino , Preescolar , Arachis/inmunología , Arachis/efectos adversos , Pruebas Cutáneas/métodos , Anafilaxia/diagnóstico , Anafilaxia/inmunología , Alérgenos/inmunología , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangre , Prueba de Estudio Conceptual , Adolescente , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Antígenos de Plantas/inmunologíaRESUMEN
BACKGROUND: Allergy to lipid transfer proteins (LPT) is common in Mediterranean Europe, and it causes severe reactions in patients and affects multiple foods, impairing the quality of life. OBJECTIVE: This study aimed to describe the clinical and sensitization profile of patients with LTP syndrome and to determine a clinical pattern of severity. Molecular diagnosis is shown in a broad population through microarrays. MATERIAL AND METHODS: This study was performed at the LTP Allergy Consultation of the Reina Sofia Hospital in Murcia, Spain. We analyzed the patients' characteristics, reactions, cofactors, food implicated, quality of life, skin prick test to food and aeroallergens, and serologic parameters, such as total immunoglobulin E, peach LTP (Pru p 3 IgE) and immunoglobulin G4, and microarray Immuno Solid-phase Allergen Chip (ISAC). We related the severity of the reactions with other variables. RESULTS: We presented a series of 236 patients diagnosed with LTP allergy, 54.66% suffering from anaphylaxis, 36.02% from urticaria angioedema, and 9.32% from oral allergy syndrome. The most frequently implicated food was peach, producing symptoms in 70% of patients, followed by walnut in 55%, peanut in 45%, hazelnut in 44%, and apple in 38% patients. Regarding the food that provoked anaphylaxis, walnut was the most frequent instigator, along with peach, peanut, hazelnut, almond, sunflower seed, and apple. According to the severity of LPT reaction, we did not discover significant differences in gender, age, food group involved, and serologic parameters. We found differences in the presence of cofactors, with 48.84% of cofactors in patients with anaphylaxis, compared to 27.1% in patients without anaphylaxis and in family allergy background (P < 0.0001). CONCLUSION: In our series of patients, 54% presented anaphylaxis, and the foods that most frequently produced symptoms were peaches, apples, and nuts. Cofactors and family allergy backgrounds were associated with the severity of LPT reaction.
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Alérgenos , Antígenos de Plantas , Hipersensibilidad a los Alimentos , Inmunoglobulina E , Pruebas Cutáneas , Humanos , Masculino , Femenino , Hipersensibilidad a los Alimentos/inmunología , Hipersensibilidad a los Alimentos/diagnóstico , Hipersensibilidad a los Alimentos/epidemiología , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Adulto , Persona de Mediana Edad , Antígenos de Plantas/inmunología , Alérgenos/inmunología , España/epidemiología , Adolescente , Proteínas de Plantas/inmunología , Adulto Joven , Proteínas Portadoras/inmunología , Niño , Inmunoglobulina G/sangre , Inmunoglobulina G/inmunología , Anciano , Calidad de Vida , Anafilaxia/inmunología , Anafilaxia/diagnóstico , Anafilaxia/etiología , PreescolarRESUMEN
Walnut and hazelnut coallergy is a frequent manifestation in clinical practice whose molecular basis remains unclear. For this purpose, walnut-hazelnut cross-reactivity was evaluated in 20 patients allergic to one or both tree nuts and sensitized to their 2S albumins. Immunoblotting assays showed that 85% of patients recognized Jug r 1, walnut 2S albumin, which was associated with the development of severe symptoms; 50% of them corecognized hazelnut 2S albumin, Cor a 14. Both allergens were isolated using chromatographic techniques. Inhibition ELISAs revealed that Jug r 1 strongly inhibited the binding of Cor a 14-specific IgE, but Cor a 14 only partially inhibited Jug r 1-specific IgE binding. Our results showed that patients sensitized to walnut/hazelnut 2S albumins were not a homogeneous population. There were patients sensitized to specific epitopes of walnut 2S albumins and patients sensitized to cross-reactive epitopes between walnut and hazelnut, with Jug r 1 being the primary sensitizer.
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Antígenos de Plantas , Corylus , Reacciones Cruzadas , Inmunoglobulina E , Juglans , Hipersensibilidad a la Nuez , Nueces , Juglans/química , Juglans/inmunología , Humanos , Corylus/química , Corylus/inmunología , Femenino , Masculino , Hipersensibilidad a la Nuez/inmunología , Adulto , Persona de Mediana Edad , Inmunoglobulina E/inmunología , Nueces/química , Nueces/inmunología , Antígenos de Plantas/inmunología , Antígenos de Plantas/química , Albuminas 2S de Plantas/inmunología , Albuminas 2S de Plantas/química , Adulto Joven , Alérgenos/inmunología , Alérgenos/química , Adolescente , Proteínas de Plantas/inmunología , Proteínas de Plantas/química , Niño , AncianoRESUMEN
Allergen detection methods support food labeling and quality assessment at the allergen component level of allergen preparations used for allergy diagnosis and immunotherapy (AIT). Commonly applied enzyme-linked immunosorbent assay (ELISA) requires animal antibodies but potentially shows batch variations. We developed synthetic aptamers as alternative binders in allergen detection to meet the replacement, reduction, and refinement (3R) principle on animal protection in science. ssDNA aptamers were specifically selected against the major peanut allergen Ara h 1 and identified by next-generation sequencing. Application in various detection systems (ELISA-like assays, western blot, and surface plasmon resonance) was demonstrated. The ELISA-like assay comprised a sensitivity of 10 ng/mL Ara h 1, comparable to published antibody-based ELISA, and allowed Ara h 1 detection in various peanut flours, similar to those used in peanut AIT as well as in processed food. This ELISA-like aptamer-based assay proofs antibody-free allergen detection for food labeling or quality assessment of diagnostic and therapeutic allergen products.
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Alérgenos , Antígenos de Plantas , Aptámeros de Nucleótidos , Arachis , Ensayo de Inmunoadsorción Enzimática , Proteínas de Plantas , Aptámeros de Nucleótidos/química , Aptámeros de Nucleótidos/inmunología , Arachis/química , Arachis/inmunología , Antígenos de Plantas/inmunología , Antígenos de Plantas/análisis , Antígenos de Plantas/genética , Proteínas de Plantas/inmunología , Proteínas de Plantas/genética , Alérgenos/inmunología , Alérgenos/análisis , Hipersensibilidad al Cacahuete/inmunología , Glicoproteínas/inmunología , Glicoproteínas/química , Proteínas de la Membrana/inmunología , Proteínas de la Membrana/genética , Humanos , Técnica SELEX de Producción de Aptámeros/métodosRESUMEN
Ragweed pollen allergy is the most common seasonal allergy in western Romania. Prolonged exposure to ragweed pollen may induce sensitization to pan-allergens such as calcium-binding proteins (polcalcins) and progression to more severe symptoms. We aimed to detect IgE sensitization to recombinant Amb a 9 and Amb a 10 in a Romanian population, to assess their potential clinical relevance and cross-reactivity, as well as to investigate the relation with clinical symptoms. rAmb a 9 and rAmb a 10 produced in Escherichia coli were used to detect specific IgE in sera from 87 clinically characterized ragweed-allergic patients in ELISA, for basophil activation experiments and rabbit immunization. Rabbit rAmb a 9- and rAmb a 10-specific sera were used to detect possible cross-reactivity with rArt v 5 and reactivity towards ragweed and mugwort pollen extracts. The results showed an IgE reactivity of 25% to rAmb a 9 and 35% to rAmb a 10. rAmb a 10 induced basophil degranulation in three out of four patients tested. Moreover, polcalcin-negative patients reported significantly more skin symptoms, whereas polcalcin-positive patients tended to report more respiratory symptoms. Furthermore, both rabbit antisera showed low reactivity towards extracts and showed high reactivity to rArt v 5, suggesting strong cross-reactivity. Our study indicated that recombinant ragweed polcalcins might be considered for molecular diagnosis.
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Proteínas de Unión al Calcio , Reacciones Cruzadas , Inmunoglobulina E , Rinitis Alérgica Estacional , Adulto , Femenino , Humanos , Masculino , Alérgenos/inmunología , Ambrosia/inmunología , Antígenos de Plantas/inmunología , Proteínas de Unión al Calcio/inmunología , Reacciones Cruzadas/inmunología , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Extractos Vegetales , Rinitis Alérgica Estacional/inmunología , Rinitis Alérgica Estacional/sangre , RumaníaRESUMEN
Pollen from Salsola kali, i.e., saltwort, Russian thistle, is a major allergen source in the coastal regions of southern Europe, in Turkey, Central Asia, and Iran. S. kali-allergic patients mainly suffer from hay-fever (i.e., rhinitis and conjunctivitis), asthma, and allergic skin symptoms. The aim of this study was to investigate the importance of individual S. kali allergen molecules. Sal k 1, Sal k 2, Sal k 3, Sal k 4, Sal k 5, and Sal k 6 were expressed in Escherichia coli as recombinant proteins containing a C-terminal hexahistidine tag and purified by nickel affinity chromatography. The purity of the recombinant allergens was analyzed by SDS-PAGE. Their molecular weight was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and their fold and secondary structure were studied by circular dichroism (CD) spectroscopy. Sera from clinically well-characterized S. kali-allergic patients were used for IgE reactivity and basophil activation experiments. S. kali allergen-specific IgE levels and IgE levels specific for the highly IgE cross-reactive profilin and the calcium-binding allergen from timothy grass pollen, Phl p 12 and Phl p 7, respectively, were measured by ImmunoCAP. The allergenic activity of natural S. kali pollen allergens was studied in basophil activation experiments. Recombinant S. kali allergens were folded when studied by CD analysis. The sum of recombinant allergen-specific IgE levels and allergen-extract-specific IgE levels was highly correlated. Sal k 1 and profilin, reactive with IgE from 64% and 49% of patients, respectively, were the most important allergens, whereas the other S. kali allergens were less frequently recognized. Specific IgE levels were highest for profilin. Of note, 37% of patients who were negative for Sal k 1 showed IgE reactivity to Phl p 12, emphasizing the importance of the ubiquitous cytoskeletal actin-binding protein, profilin, for the diagnosis of IgE sensitization in S. kali-allergic patients. rPhl p 12 and rSal k 4 showed equivalent IgE reactivity, and the clinical importance of profilin was underlined by the fact that profilin-monosensitized patients suffered from symptoms of respiratory allergy to saltwort. Accordingly, profilin should be included in the panel of allergen molecules for diagnosis and in molecular allergy vaccines for the treatment and prevention of S. kali allergy.
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Alérgenos , Reacciones Cruzadas , Inmunoglobulina E , Polen , Profilinas , Salsola , Humanos , Profilinas/inmunología , Profilinas/química , Inmunoglobulina E/inmunología , Alérgenos/inmunología , Alérgenos/genética , Salsola/inmunología , Femenino , Polen/inmunología , Masculino , Reacciones Cruzadas/inmunología , Adulto , Proteínas Recombinantes/inmunología , Rinitis Alérgica Estacional/inmunología , Persona de Mediana Edad , Basófilos/inmunología , Basófilos/metabolismo , Antígenos de Plantas/inmunología , Antígenos de Plantas/genética , Adulto Joven , Adolescente , Proteínas de Plantas/inmunología , Proteínas de Plantas/genéticaRESUMEN
BACKGROUND AND OBJECTIVE: Sensitization to Blomia tropicalis is associated with asthma in various tropical and subtropical countries; however, information about the specific molecular components associated with this disease is scarce. Using molecular diagnosis, we sought to identify B tropicalis allergens associated with asthma in Colombia. METHODS: Specific IgE (sIgE) to 8 B tropicalis recombinant allergens (Blo t 2, 5, 7, 8, 10, 12, 13, and 21) was determined using an in-house ELISA system in asthma patients (n=272) and controls (n=298) recruited in a national prevalence study performed in several Colombian cities (Barranquilla, Bogotá, Medellín, Cali, and San Andrés). The study sample included children and adults (mean [SD] age, 28 [17] years). Cross-reactivity between Blo t 5 and Blo t 21 was evaluated using ELISA-inhibition. RESULTS: Specific IgE (sIgE) to 8 B tropicalis recombinant allergens (Blo t 2, 5, 7, 8, 10, 12, 13, and 21) was determined using an in-house ELISA system in asthma patients (n=272) and controls (n=298) recruited in a national prevalence study performed in several Colombian cities (Barranquilla, Bogotá, Medellín, Cali, and San Andrés). The study sample included children and adults (mean [SD] age, 28 [17] years). Cross-reactivity between Blo t 5 and Blo t 21 was evaluated using ELISA-inhibition. CONCLUSION: Although Blo t 5 and Blo t 21 are considered common sensitizers, this is the first report of their association with asthma. Both components should be included in molecular panels for diagnosis of allergy in the tropics.
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Alérgenos , Asma , Inmunoglobulina E , Humanos , Asma/inmunología , Asma/diagnóstico , Asma/epidemiología , Inmunoglobulina E/inmunología , Inmunoglobulina E/sangre , Adulto , Masculino , Femenino , Estudios de Casos y Controles , Niño , Adolescente , Colombia/epidemiología , Alérgenos/inmunología , Adulto Joven , Persona de Mediana Edad , Antígenos de Plantas/inmunología , Reacciones Cruzadas , Clima Tropical , Prevalencia , PreescolarRESUMEN
Food allergies mediated by specific IgE (sIgE) have a significant socioeconomic impact on society. Evaluating the IgE cross-reactivity between allergens from different allergen sources can enable the better management of these potentially life-threatening adverse reactions to food proteins and enhance food safety. A novel banana fruit allergen, S-adenosyl-L-homocysteine hydrolase (SAHH), has been recently identified and its recombinant homolog was heterologously overproduced in E. coli. In this study, we performed a search in the NCBI (National Center for Biotechnology Information) for SAHH homologs in ryegrass, latex, and kiwifruit, all of which are commonly associated with pollen-latex-fruit syndrome. In addition, Western immunoblot analysis was utilized to identify the cross-reactive IgE to banana SAHH in the sera of patients with a latex allergy, kiwifruit allergy, and ryegrass allergy. ClustalOmega analysis showed more than 92% amino acid sequence identity among the banana SAHH homologs in ryegrass, latex, and kiwifruit. In addition to five B-cell epitopes, in silico analysis predicted eleven T-cell epitopes in banana SAHH, seventeen in kiwifruit SAHH, twelve in ryegrass SAHH, and eight in latex SAHH, which were related to the seven-allele HLA reference set (HLA-DRB1*03:01, HLA-DRB1*07:01, HLA-DRB1*15:01, HLA-DRB3*01:01, HLA-DRB3*02:02, HLA-DRB4*01:01, HLA-DRB5*01:01). Four T-cell epitopes were identical in banana and kiwifruit SAHH (positions 328, 278, 142, 341), as well as banana and ryegrass SAHH (positions 278, 142, 96, and 341). All four SAHHs shared two T-cell epitopes (positions 278 and 341). In line with the high amino acid sequence identity and B-cell epitope homology among the analyzed proteins, the cross-reactive IgE to banana SAHH was detected in three of three latex-allergic patients, five of six ryegrass-allergic patients, and two of three kiwifruit-allergic patients. Although banana SAHH has only been studied in a small group of allergic individuals, it is a novel cross-reactive food allergen that should be considered when testing for pollen-latex-fruit syndrome.
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Actinidia , Alérgenos , Reacciones Cruzadas , Hipersensibilidad a los Alimentos , Inmunoglobulina E , Látex , Musa , Humanos , Reacciones Cruzadas/inmunología , Hipersensibilidad a los Alimentos/inmunología , Alérgenos/inmunología , Alérgenos/genética , Musa/inmunología , Musa/genética , Inmunoglobulina E/inmunología , Actinidia/inmunología , Femenino , Látex/inmunología , Masculino , Proteínas de Plantas/inmunología , Proteínas de Plantas/genética , Adulto , Antígenos de Plantas/inmunología , Antígenos de Plantas/genética , Secuencia de Aminoácidos , Epítopos de Linfocito T/inmunología , Persona de Mediana Edad , Adolescente , Niño , Adulto JovenRESUMEN
Pollen from common ragweed is an important allergen source worldwide and especially in western and southern Romania. More than 100 million patients suffer from symptoms of respiratory allergy (e.g., rhinitis, asthma) to ragweed pollen. Among the eleven characterized allergens, Amb a 6 is a non-specific lipid transfer protein (nsLTP). nsLTPs are structurally stable proteins in pollen and food from different unrelated plants capable of inducing severe reactions. The goal of this study was to produce Amb a 6 as a recombinant and structurally folded protein (rAmb a 6) and to characterize its physicochemical and immunological features. rAmb a 6 was expressed in Spodoptera frugiperda Sf9 cells as a secreted protein and characterized by mass spectrometry and circular dichroism (CD) spectroscopy regarding molecular mass and fold, respectively. The IgE-binding frequency towards the purified protein was evaluated using sera from 150 clinically well-characterized ragweed-allergic patients. The allergenic activities of rAmb a 6 and the nsLTP from the weed Parietaria judaica (Par j 2) were evaluated in basophil activation assays. rAmb a 6-specific IgE reactivity was associated with clinical features. Pure rAmb a 6 was obtained by insect cell expression. Its deduced molecular weight corresponded to that determined by mass spectrometry (i.e., 10,963 Da). rAmb a 6 formed oligomers as determined by SDS-PAGE under non-reducing conditions. According to multiple sequence comparisons, Amb a 6 was a distinct nsLTP with less than 40% sequence identity to currently known plant nsLTP allergens, except for nsLTP from Helianthus (i.e., 52%). rAmb a 6 is an important ragweed allergen recognized by 30% of ragweed pollen allergic patients. For certain patients, rAmb a 6-specific IgE levels were higher than those specific for the major ragweed allergen Amb a 1 and analysis also showed a higher allergenic activity in the basophil activation test. rAmb a 6-positive patients suffered mainly from respiratory symptoms. The assumption that Amb a 6 is a source-specific ragweed allergen is supported by the finding that none of the patients showing rAmb a 6-induced basophil activation reacted with Par j 2 and only one rAmb a 6-sensitized patient had a history of plant food allergy. Immunization of rabbits with rAmb a 6 induced IgG antibodies which strongly inhibited IgE binding to rAmb a 6. Our results demonstrate that Amb a 6 is an important source-specific ragweed pollen allergen that should be considered for diagnosis and allergen-specific immunotherapy of ragweed pollen allergy.