Person: Batanero Cremades, Eva
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First Name
Eva
Last Name
Batanero Cremades
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Bioquímica y Biología Molecular
Area
Bioquímica y Biología Molecular
Identifiers
8 results
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Now showing 1 - 8 of 8
Item Characterization of Relevant Biomarkers for the Diagnosis of Food Allergies: An Overview of the 2S Albumin Family(Foods, 2021) Bueno Díaz, Cristina; Martín-Pedraza, Laura; Parrón Ballesteros, Jorge; Cuesta-Herranz, Javier; Cabanillas, Beatriz; Pastor Vargas, Carlos; Batanero Cremades, Eva; Villalba Díaz, María Teresa2S albumins are relevant and often major allergens from several tree nuts and seeds, affecting mainly children and young people. The present study aims to assess how the structural features of 2S albumins could affect their immunogenic capacity, which is essential to comprehend the role of these proteins in food allergy. For this purpose, twelve 2S albumins were isolated from their respective extracts by chromatographic methods and identified by MALDI-TOF massspectrometry. Their molecular and structural characterization was conducted by electrophoretic, spectroscopic and in silico methods, showing that these are small proteins that comprise a wide range of isoelectric points, displaying a general high structure stability to thermal treatment. Despite low amino acid sequence identity, these proteins share structural features, pointing conformational epitopes to explain cross-reactivity between them. Immunoblotting with allergic patients’ sera revealed those possible correlations between evolutionarily distant 2S albumins from different sources. The availability of a well-characterized panel of 2S albumins from plant-derived sources allowed establishing correlations between their structural features and their allergenic potential, including their role in cross-reactivity processesItem 2S albumins and 11S globulins, two storage proteins involved in pumpkin seeds allergy(Allergy: European Journal of Allergy and Clinical Immunology, 2021) Bueno Díaz, Cristina; Martín-Pedraza, Laura; León, Laura; Haroun-Díaz, Elisa; Pastor Vargas, Carlos; Muñoz-García, Esther; De las Heras, Manuel; Batanero Cremades, Eva; Cuesta-Herranz, Javier; Villalba Díaz, María TeresaMembers from Cucurbitaceae family have been reported to induce food allergy.1, 7 Pumpkin (Cucurbita maxima) pulp has been mostly the allergenic source but few studies are focused on the allergenic potential of its seeds.2 Pumpkin seed may be consumed as snacks or as components in other food products, becoming hidden allergens, eliciting infrequent but severe cases of allergy with life-threatening reactions.3 The nature of those allergens has not been investigated in detail so far. This study aimed to identify two allergens, from pumpkin seeds a 2S albumin and an 11S globulin, involved in severe allergic reactions of four patients allergic to these seeds and evaluate the cross-reactivity with other seeds and nuts containing homologous proteins. General characteristics of selected individuals, extracted from their clinical histories, are shown in Table S1. All patients described immediate allergic reactions with severe and systemic symptoms as anaphylaxis, showing a positive specific IgE (ImmunoCAP, Thermo Fisher) and Skin prick testing (SPT) to pumpkin seeds extractItem Characterization of Profilin and Polcalcin Panallergens From Ash Pollen(Journal of Investigational Allergology and Clinical Immunology, 2014) Mas García, Salvador; Garrido Arandia, María; Batanero Cremades, Eva; Purohit, A.; Pauli, G.; Rodríguez García, Rosalía; Barderas Manchado, Rodrigo; Villalba Díaz, María TeresaBackground: Ash (Fraxinus excelsior) is an important source of allergenic pollen in temperate areas of Europe. Profilin and polcalcin are 2 important panallergens involved in cross-reactivity between different sources. Objective: To clone and produce Fra e 2 (profilin) and Fra e 3 (polcalcin) as recombinant proteins and evaluate their immunological properties using the natural forms obtained from ash pollen. Methods: Total RNA from ash pollen was used as a template to obtain the specific complementary DNA (cDNA) sequences of the 2 panallergens. The cDNA-encoding sequences were cloned into the pET11b expression vector and used to transform BL21 (DE3) Escherichia coli cells. Proteins were expressed, purified by chromatography, and characterized structurally by circular dichroism, mass spectrometry, and immunologically by western blot and ELISA using profilin and polcalcin polyclonal antibodies and human sera from ash pollen–sensitized patients. Results: Profilin and polcalcin amino acid sequences from ash pollen showed a high degree of identity with homologous allergens from different sources. The cDNA-encoding allergen sequences were expressed as nonfusion recombinant proteins and purified to homogeneity. Secondary structure values were similar to those obtained from other members of these families. Allergenic Properties of the recombinant allergens were observed to be equivalent to those of the natural counterparts of F excelsior pollen.Conclusions: Fra e 2 and Fra e 3 recombinant allergens might be used in clinical diagnosis to determine profilin- and polcalcin-specific IgE levels present in the sera of ash pollen–sensitized patients, thus facilitating the finding of the sensitizing source in areas with complex sensitization profiles.Item A Deletion Variant of the Aspergillus fumigatus Ribotoxin Asp f 1 Induces an Attenuated Airway Inflammatory Response in a Mouse Model of Sensitization(Journal of Investigational Allergology & Clinical Immunology, 2010) Álvarez García, Elisa; Batanero Cremades, Eva; García Fernández, Rosa Ana; Villalba Díaz, María Teresa; Gavilanes, José G.; Martínez Del Pozo, Álvaroα-Sarcin is a natural variant of Asp f 1 produced by the nonpathogenic fungus Aspergillus giganteus. Both proteins show a sequence identity of 87% and almost identical 3-dimensional structures. α-Sarcin Δ(7-22) is a deletion mutant that displays reduced immunoglobulin (Ig) E reactivity and is much less cytotoxic than wild-type proteins against human transformed cells. Objective: A murine model of sensitization to Asp f 1 was established to test the response elicited by this α-sarcin Δ(7-22) deletion mutant. Methods: BALB/c mice were treated intraperitoneally with different mixtures of recombinant wild-type Asp f 1 and/or a suspension of a commercially available A fumigatus standard extract. Mice were then intranasally challenged with Asp f 1 or α-sarcin Δ(7-22). Sera were collected for subsequent measurement of Ig levels and histological analysis of the nostrils and lungs. Results: Sensitization to Asp f 1 was successful only when the purifi ed protein was fi rst administered together with the A fumigatus suspension. The model was characterized by elevated levels of total IgE in serum and histological lesions in the lungs and nostrils. These symptoms were less severe when the deletion variant was the protein administered, thus confi rming in vivo its lower toxic character. Conclusions: An easily reproducible mouse model of A fumigatus Asp f 1 sensitization was established. This model revealed α-sarcin Δ (7-22) to be a potential candidate for immunotherapy.Item Pollensomes as natural vehicles for pollen allergens(Journal of Inmunology, 2015) Prado, Noela; De Linares, C.; Sanz, M.L.; Gamboa, P.; Villalba Díaz, María Teresa; Rodríguez García, Rosalía; Batanero Cremades, EvaOlive (Olea europaea) pollen constitutes one of the most important allergen sources in the Mediterranean countries and some areas of the United States, South Africa, and Australia. Recently, we provided evidence that olive pollen releases nanovesicles of respirable size, named generically pollensomes, during in vitro germination. Olive pollensomes contain allergens, such as Ole e 1, Ole e 11, and Ole e 12, suggesting a possible role in allergy. The aim of this study was to assess the contribution of pollensomes to the allergic reaction. We show that pollensomes exhibit allergenic activity in terms of patients' IgE-binding capacity, human basophil activation, and positive skin reaction in sensitized patients. Furthermore, allergen-containing pollensomes have been isolated from three clinically relevant nonphylogenetically related species: birch (Betula verrucosa), pine (Pinus sylvestris), and ryegrass (Lolium perenne). Most interesting, pollensomes were isolated from aerobiological samples collected with an eight-stage cascade impactor collector, indicating that pollensomes secretion is a naturally occurring phenomenon. Our findings indicate that pollensomes may represent widespread vehicles for pollen allergens, with potential implications in the allergic reaction.Item Allium porrum Extract Decreases Effector Cell Degranulation and Modulates Airway Epithelial Cell Function(Nutrients, 2019) Benedé Pérez, Sara; Gradillas, Ana; Villalba Díaz, María Teresa; Batanero Cremades, EvaAllium genus plants, such as leek (Allium porrum), are rich sources of anti-inflammatory and anti-oxidant secondary metabolites; this is of interest because it demonstrates their suitability as pharmacological alternatives for inflammatory processes, including allergy treatment. The composition of methanolic leek extract (LE) was analyzed by GC–MS and LC–IT/MS, and the total phenolic content and antioxidant capacity were quantified by colorimetric methods. Its pharmacological potential was analyzed in human bronchial epithelial Calu-3 cells, human mast cells LAD2, and humanized rat basophiles RBL-2H3. LE exhibited a cytotoxic effect on Calu-3 cells and HumRBL-2H3 cells only at high concentrations and in a dose-dependent manner. Moreover, LE decreased the degranulation of LAD2 and HumRBL-2H3 cells. LE treatment also significantly prevented alterations in transepithelial electrical resistance values and mRNA levels of glutathione-S-transferase (GST), c-Jun, and NFκB after treatment with H2O2 in ALI-cultured Calu-3 cells. Finally, ALI-cultured Calu-3 cells treated with LE showed lower permeability to Ole e 1 compared to untreated cells. A reduction in IL-6 secretion in ALI-cultured Calu-3 cells treated with LE was also observed. In summary, the results obtained in this work suggest that A. porrum extract may have potential anti-allergic effects due to its antioxidant and anti-inflammatory properties. This study provides several important insights into how LE can protect against allergy.Item Ash pollen immunoproteomics: Identification, immunologic characterization, and sequencing of 6 new allergens(The Journal of Allergy and Clinical Inmunology, 2014) Mas García, Salvador; Torres Pardo, María De La Almudena; Garrido-Arandia, María; Salamanca, Guillermo; Castro, Lourdes; Barral, Patricia; Purohit, Ashok; Pauli, Gabrielle; Rodríguez García, Rosalía; Batanero Cremades, Eva; Barderas Manchado, Rodrigo; Villalba Díaz, María TeresaImmunoproteomics, IgE-inhibition assays and cDNA-cloning reveals that ash and olive allergenic protein profiles are mostly equivalent, thus explaining their high cross reactivity. Our data suggest simplifying diagnosis of patients by using indistinctly ash or olive pollen.Item Beyond allergic progression: From molecules to microbes as barrier modulators in the gut-lung axis functionality(Frontiers in Allergy, 2023) Parrón Ballesteros, Jorge; Rubén García Gordo; López Rodríguez, Juan Carlos; Olmo López, Nieves; Villalba Díaz, María Teresa; Batanero Cremades, Eva; Turnay Abad, Francisco JavierThe “epithelial barrier hypothesis” states that a barrier dysfunction can result in allergy development due to tolerance breakdown. This barrier alteration may come from the direct contact of epithelial and immune cells with the allergens, and indirectly, through deleterious effects caused by environmental changes triggered by industrialization, pollution, and changes in the lifestyle. Apart from their protective role, epithelial cells can respond to external factors secreting IL-25 IL-33, and TSLP, provoking the activation of ILC2 cells and a Th2-biased response. Several environmental agents that influence epithelial barrier function, such as allergenic proteases, food additives or certain xenobiotics are reviewed in this paper. In addition, dietary factors that influence the allergenic response in a positive or negative way will be also described here. Finally, we discuss how the gut microbiota, its composition, and microbe-derived metabolites, such as short-chain fatty acids, alter not only the gut but also the integrity of distant epithelial barriers, focusing this review on the gut-lung axis.