Browsing by Author "Kuehn, Annette"
Now showing 1 - 9 of 9
Results Per Page
Sort Options
- Are Physicochemical Properties Shaping the Allergenic Potency of Animal Allergens?Publication . Costa, Joana; Villa, Caterina; Verhoeckx, Kitty; Cirkovic-Velickovic, Tanja; Schrama, Denise; Roncada, Paola; Rodrigues, Pedro M.; Piras, Cristian; Martin-Pedraza, Laura; Monaci, Linda; Molina, Elena; Mazzucchelli, Gabriel; Mafra, Isabel; Lupi, Roberta; Lozano-Ojalvo, Daniel; Larre, Colette; Klueber, Julia; Gelencser, Eva; Bueno-Diaz, Cristina; Diaz-Perales, Araceli; Benede, Sara; Bavaro, Simona Lucia; Kuehn, Annette; Hoffmann-Sommergruber, Karim; Holzhauser, ThomasKey determinants for the development of an allergic response to an otherwise 'harmless' food protein involve different factors like the predisposition of the individual, the timing, the dose, the route of exposure, the intrinsic properties of the allergen, the food matrix (e.g. lipids) and the allergen modification by food processing. Various physicochemical parameters can have an impact on the allergenicity of animal proteins. Following our previous review on how physicochemical parameters shape plant protein allergenicity, the same analysis was proceeded here for animal allergens. We found that each parameter can have variable effects, ranging on an axis from allergenicity enhancement to resolution, depending on its nature and the allergen. While glycosylation and phosphorylation are common, both are not universal traits of animal allergens. High molecular structures can favour allergenicity, but structural loss and uncovering hidden epitopes can also have a similar impact. We discovered that there are important knowledge gaps in regard to physicochemical parameters shaping protein allergenicity both from animal and plant origin, mainly because the comparability of the data is poor. Future biomolecular studies of exhaustive, standardised design together with strong validation part in the clinical context, together with data integration model systems will be needed to unravel causal relationships between physicochemical properties and the basis of protein allergenicity.
- Are physicochemical properties shaping the allergenic potency of plant allergens?Publication . Costa, Joana; Bavaro, Simona Lucia; Benede, Sara; Diaz-Perales, Araceli; Bueno-Diaz, Cristina; Gelencser, Eva; Klueber, Julia; Larre, Colette; Lozano-Ojalvo, Daniel; Lupi, Roberta; Mafra, Isabel; Mazzucchelli, Gabriel; Molina, Elena; Monaci, Linda; Martin-Pedraza, Laura; Piras, Cristian; Rodrigues, Pedro M.; Roncada, Paola; Schrama, Denise; Cirkovic-Velickovic, Tanja; Verhoeckx, Kitty; Villa, Caterina; Kuehn, Annette; Hoffmann-Sommergruber, Karin; Holzhauser, ThomasThis review searched for published evidence that could explain how different physicochemical properties impact on the allergenicity of food proteins and if their effects would follow specific patterns among distinct protein families. Owing to the amount and complexity of the collected information, this literature overview was divided in two articles, the current one dedicated to protein families of plant allergens and a second one focused on animal allergens. Our extensive analysis of the available literature revealed that physicochemical characteristics had consistent effects on protein allergenicity for allergens belonging to the same protein family. For example, protein aggregation contributes to increased allergenicity of 2S albumins, while for legumins and cereal prolamins, the same phenomenon leads to a reduction. Molecular stability, related to structural resistance to heat and proteolysis, was identified as the most common feature promoting plant protein allergenicity, although it fails to explain the potency of some unstable allergens (e.g. pollen-related food allergens). Furthermore, data on physicochemical characteristics translating into clinical effects are limited, mainly because most studies are focused on in vitro IgE binding. Clinical data assessing how these parameters affect the development and clinical manifestation of allergies is minimal, with only few reports evaluating the sensitising capacity of modified proteins (addressing different physicochemical properties) in murine allergy models. In vivo testing of modified pure proteins by SPT or DBPCFC is scarce. At this stage, a systematic approach to link the physicochemical properties with clinical plant allergenicity in real-life scenarios is still missing.
- EAACI molecular allergology user's guide 2.0Publication . Dramburg, Stephanie; Hilger, Christiane; Santos, Alexandra F.; de las Vecillas, Leticia; Aalberse, Rob C.; Acevedo, Nathalie; Aglas, Lorenz; Altmann, Friedrich; Arruda, Karla L.; Asero, Riccardo; Ballmer‐Weber, Barbara; Barber, Domingo; Beyer, Kirsten; Biedermann, Tilo; Bilo, Maria Beatrice; Blank, Simon; Bosshard, Philipp P.; Breiteneder, Heimo; Brough, Helen A.; Bublin, Merima; Campbell, Dianne; Caraballo, Luis; Caubet, Jean Christoph; Celi, Giorgio; Chapman, Martin D.; Chruszcz, Maksymilian; Custovic, Adnan; Czolk, Rebecca; Davies, Janet; Douladiris, Nikolaos; Eberlein, Bernadette; Ebisawa, Motohiro; Ehlers, Anna; Eigenmann, Philippe; Gadermaier, Gabriele; Giovannini, Mattia; Gomez, Francisca; Grohman, Rebecca; Guillet, Carole; Hafner, Christine; Hamilton, Robert G.; Hauser, Michael; Hawranek, Thomas; Hoffmann, Hans Jürgen; Holzhauser, Thomas; Iizuka, Tomona; Jacquet, Alain; Jakob, Thilo; Janssen‐Weets, Bente; Jappe, Uta; Jutel, Marek; Kalic, Tanja; Kamath, Sandip; Kespohl, Sabine; Kleine‐Tebbe, Jörg; Knol, Edward; Knulst, André; Konradsen, Jon R.; Korošec, Peter; Kuehn, Annette; Lack, Gideon; Le, Thuy‐My; Lopata, Andreas; Luengo, Olga; Mäkelä, Mika; Marra, Alessandro Maria; Mills, Clare; Morisset, Martine; Muraro, Antonella; Nowak‐Wegrzyn, Anna; Nugraha, Roni; Ollert, Markus; Palosuo, Kati; Pastorello, Elide Anna; Patil, Sarita Ulhas; Platts‐Mills, Thomas; Pomés, Anna; Poncet, Pascal; Potapova, Ekaterina; Poulsen, Lars K.; Radauer, Christian; Radulovic, Suzana; Raulf, Monika; Rougé, Pierre; Sastre, Joaquin; Sato, Sakura; Scala, Enrico; Schmid, Johannes M.; Schmid‐Grendelmeier, Peter; Schrama, Denise; Sénéchal, Hélène; Traidl‐Hoffmann, Claudia; Valverde‐Monge, Marcela; van Hage, Marianne; van Ree, Ronald; Verhoeckx, Kitty; Vieths, Stefan; Wickman, Magnus; Zakzuk, Josefina; Matricardi, Paolo M.; Hoffmann‐Sommergruber, KarinSince the discovery of immunoglobulin E (IgE) as a mediator of allergic diseases in 1967, our knowledge about the immunological mechanisms of IgE-mediated allergies has remarkably increased. In addition to understanding the immune response and clinical symptoms, allergy diagnosis and management depend strongly on the precise identification of the elicitors of the IgE-mediated allergic reaction. In the past four decades, innovations in bioscience and technology have facilitated the identification and production of well-defined, highly pure molecules for component-resolved diagnosis (CRD), allowing a personalized diagnosis and management of the allergic disease for individual patients. The first edition of the "EAACI Molecular Allergology User's Guide" (MAUG) in 2016 rapidly became a key reference for clinicians, scientists, and interested readers with a background in allergology, immunology, biology, and medicine. Nevertheless, the field of molecular allergology is moving fast, and after 6 years, a new EAACI Taskforce was established to provide an updated document. The Molecular Allergology User's Guide 2.0 summarizes state-of-the-art information on allergen molecules, their clinical relevance, and their application in diagnostic algorithms for clinical practice. It is designed for both, clinicians and scientists, guiding health care professionals through the overwhelming list of different allergen molecules available for testing. Further, it provides diagnostic algorithms on the clinical relevance of allergenic molecules and gives an overview of their biology, the basic mechanisms of test formats, and the application of tests to measure allergen exposure.
- Effect of creatine and EDTA supplemented diets on European seabass (Dicentrarchus labrax) allergenicity, fish muscle quality and omics fingerprintPublication . Schrama, Denise; Raposo de Magalhães, Cláudia; Cerqueira, Marco; Carrilho, Raquel; Farinha, Ana Paula; Costa, Ana Rosa da; Gonçalves, Amparo; Kuehn, Annette; Revets, Dominique; Planchon, Sébastien; Engrola, Sofia; Rodrigues, PedroThe relatively easy access to fish worldwide, alongside the increase of aquaculture production contributes to increased fish consumption which result in higher prevalence of respective allergies. Allergies to fish constitute a significant concern worldwide. beta-parvalbumin is the main elicitor for IgE-mediated reactions. Creatine, involved in the muscle energy metabolism, and ethylenediamine tetraacetic acid (EDTA), a calcium chelator, are potential molecules to modulate parvalbumin. The purpose of this study was to test creatine (2, 5 and 8%) and EDTA (1.5, 3 and 4.5%) supplementation in fish diets to modulate beta-parvalbumin expression and structure and its allergenicity in farmed European seabass (Dicentrarchus labrax) while assessing its effects on the end-product quality. Fish welfare and muscle quality parameters were evaluated by plasma metabolites, rigor mortis, muscle pH and sensory and texture analysis. Proteomics was used to assess alterations in muscle proteome profile and metabolic fingerprinting by Fourier transform infrared spectroscopy was used to assess the liver metabolic profile. In addition, IgE-reactivity to parvalbumin was analysed using fish allergic patient sera. Metabolic fingerprinting of liver tissue revealed no major alterations in infrared spectra with creatine supplementation, while with EDTA, only absorption bands characteristic of lipids were altered. Comparative proteomics showed up regulation of (tropo) myosin and phosphoglycerate mutase 2 with Creatine supplementation. In the case of EDTA proteomics showed up regulation of proteins involved in cellular and ion homeostasis. Allergenicity seems not to be modulated with creatine or EDTA supplementation as no decreased expression levels were found and IgE-binding reactivity showed no quantitative differences.
- Effect of EDTA enriched diets on farmed fish allergenicity and muscle quality; a proteomics approachPublication . Raposo de Magalhães, Cláudia; Schrama, Denise; Fonseca, Flavio; Kuehn, Annette; Morisset, Martine; Ferreira, Sara; Goncalves, Amparo; Rodrigues, PedroFish is one of the most common elicitors of food-allergic reactions worldwide. These reactions are triggered by the calcium-binding muscle protein beta-parvalbumin, which was shown to have reduced immunoglobulin E (IgE)-binding capacity upon calcium depletion. This work aimed to reduce gilthead seabream allergenicity using diets supplemented with a calcium chelator. Three experimental feeds were tested, differing in ethylenediaminetetraacetic acid (EDTA) supplementation, and its effects on muscle and parvalbumin's IgE-reactivity were analyzed. Chromatographic determination of EDTA showed no accumulation in the muscle and sensory results demonstrated that the lowest concentration did not affect fish quality as edible fish. Proteomics revealed one protein related to muscle contraction with significantly different relative abundance. Immunoblot assays performed with fish-allergic patients sera indicated a 50% reduction in IgE-reactivity upon EDTA presence. These preliminary results provide the basis for the further development of a non-GMO approach to modulate fish allergenicity and improve safety of aquaculture fish.
- Fish allergenicity modulation using tailored enriched diets—Where are we?Publication . Schrama, Denise; Czolk, Rebecca; Raposo de Magalhães, Cláudia; Kuehn, Annette; Rodrigues, PedroFood allergy is an abnormal immune response to specific proteins in a certain food. The chronicity, prevalence, and the potential fatality of food allergy, make it a serious socio-economic problem. Fish is considered the third most allergenic food in the world, affecting part of the world population with a higher incidence in children and adolescents. The main allergen in fish, responsible for the large majority of fish-allergic reactions in sensitized patients, is a small and stable calcium-binding muscle protein named beta-parvalbumin. Targeting the expression or/and the 3D conformation of this protein by adding specific molecules to fish diets has been the innovative strategy of some researchers in the fields of fish allergies and nutrition. This has shown promising results, namely when the apo-form of beta-parvalbumin is induced, leading in the case of gilthead seabream to a 50% reduction of IgE-reactivity in fish allergic patients.
- Fish allergy management: from component-resolved diagnosis to unmet diagnostic needsPublication . Klueber, Julia; Schrama, Denise; Rodrigues, Pedro; Dickel, Heinrich; Kuehn, AnnettePurpose of review Fish is a common elicitor of IgE-mediated food allergy. Fish includes a large variety of foods, in terms of species and food processing, with marked distinction in local diets around the globe. Fish-allergic patients present with phenotypic diversity and major differences in levels of clinical cross-reactivity, features that pose an important challenge for the clinical diagnosis and management. Recent findings Parvalbumin is the major fish allergen. However, a single molecule is not sufficient but several homologs, allergens different from parvalbumin and allergen extracts, are needed for IgE-based diagnosis. Summary Parvalbumin-specific IgE are markers for clinical cross-reactions. Added value is provided by IgE typing to parvalbumin homologs from distantly related fish. IgE cosensitization profiles (parvalbumin, enolase, aldolase) are referred as severity markers. The allergen panel seems to be not yet complete why fish extracts still play a crucial role inserum IgE analysis. Further clinical validation of a multiplex approach in molecular fish allergy diagnosis is needed for striving to avoid unnecessary food restrictions and in a further sense, improved patient care.
- Fish processing and digestion affect parvalbumins detectability in Gilthead Seabream and European seabassPublication . Schrama, Denise; Raposo de Magalhães, Cláudia; Cerqueira, Marco; Carrilho, Raquel; Revets, Dominique; Kuehn, Annette; Engrola, Sofia; Rodrigues, PedroConsumption of aquatic food, including fish, accounts for 17% of animal protein intake. However, fish consumption might also result in several side-effects such as sneezing, swelling and anaphylaxis in sensitized consumers. Fish allergy is an immune reaction to allergenic proteins in the fish muscle, for instance parvalbumin (PV), considered the major fish allergen. In this study, we characterize PV in two economically important fish species for southern European aquaculture, namely gilthead seabream and European seabass, to understand its stability during in vitro digestion and fish processing. This information is crucial for future studies on the allergenicity of processed fish products. PVs were extracted from fish muscles, identified by mass spectrometry (MS), and detected by sandwich enzyme-linked immunosorbent assay (ELISA) after simulated digestion and various food processing treatments. Secondary structures were determined by circular dichroism (CD) after purification by anion exchange and gel filtration chromatography. In both species, PVs presented as α-helical and β-sheet structures, at room temperature, were shown to unfold at boiling temperatures. In European seabass, PV detectability decreased during the simulated digestion and after 240 min (intestinal phase) no detection was observed, while steaming showed a decrease (p < 0.05) in PVs detectability in comparison to raw muscle samples, for both species. Additionally, freezing (−20 °C) for up to 12 months continued to reduce the detectability of PV in tested processing techniques. We concluded that PVs from both species are susceptible to digestion and processing techniques such as steaming and freezing. Our study obtained preliminary results for further research on the allergenic potential of PV after digestion and processing.
- Protein changes as robust signatures of fish chronic stress: a proteomics approach to fish welfare researchPublication . Raposo de Magalhães, Cláudia; Schrama, Denise; Farinha, Ana Paula; Revets, Dominique; Kuehn, Annette; Planchon, Sébastien; Rodrigues, Pedro; Cerqueira, MarcoBackground Aquaculture is a fast-growing industry and therefore welfare and environmental impact have become of utmost importance. Preventing stress associated to common aquaculture practices and optimizing the fish stress response by quantification of the stress level, are important steps towards the improvement of welfare standards. Stress is characterized by a cascade of physiological responses that, in-turn, induce further changes at the whole-animal level. These can either increase fitness or impair welfare. Nevertheless, monitorization of this dynamic process has, up until now, relied on indicators that are only a snapshot of the stress level experienced. Promising technological tools, such as proteomics, allow an unbiased approach for the discovery of potential biomarkers for stress monitoring. Within this scope, using Gilthead seabream (Sparus aurata) as a model, three chronic stress conditions, namely overcrowding, handling and hypoxia, were employed to evaluate the potential of the fish protein-based adaptations as reliable signatures of chronic stress, in contrast with the commonly used hormonal and metabolic indicators. Results A broad spectrum of biological variation regarding cortisol and glucose levels was observed, the values of which rose higher in net-handled fish. In this sense, a potential pattern of stressor-specificity was clear, as the level of response varied markedly between a persistent (crowding) and a repetitive stressor (handling). Gel-based proteomics analysis of the plasma proteome also revealed that net-handled fish had the highest number of differential proteins, compared to the other trials. Mass spectrometric analysis, followed by gene ontology enrichment and protein-protein interaction analyses, characterized those as humoral components of the innate immune system and key elements of the response to stimulus. Conclusions Overall, this study represents the first screening of more reliable signatures of physiological adaptation to chronic stress in fish, allowing the future development of novel biomarker models to monitor fish welfare.