Batanero Cremades, Eva

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Last Name
Batanero Cremades
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Bioquímica y Biología Molecular
Bioquímica y Biología Molecular
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Now showing 1 - 9 of 9
  • Publication
    Characterization of Profilin and Polcalcin Panallergens From Ash Pollen
    (Esmon Publicidad, 2014) Mas García, Salvador; Garrido Arandia, María; Batanero Cremades, Eva; Purohit, A.; Pauli, G.; Rodríguez García, Rosalía; Barderas Manchado, Rodrigo; Villalba Díaz, Mayte
    Background: 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.
  • Publication
    Pollensomes as natural vehicles for pollen allergens
    (American Association of Immunologists, 2015-07-15) Prado, Noela; De Linares, C.; Sanz, M.L.; Gamboa, P.; Villalba Díaz, Mayte; Rodríguez García, Rosalía; Batanero Cremades, Eva
    Olive (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.
  • Publication
    A Deletion Variant of the Aspergillus fumigatus Ribotoxin Asp f 1 Induces an Attenuated Airway Inflammatory Response in a Mouse Model of Sensitization
    (Esmon Publicidad, 2010-03) Álvarez García, Elisa; Batanero Cremades, Eva; García Fernández, Rosa; Villalba Díaz, Mayte; Gavilanes, José G.; Martínez del Pozo, Álvaro
    α-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.
  • Publication
    Química Inspirada por la Naturaleza: Lecciones en el Museo Nacional de Ciencias Naturales
    (2017) Batanero Cremades, Eva; Yélamos López, Belén; Fernández Fernández, Mª Inmaculada; López García-Gallo, Pilar; López Rodríguez, Juan Carlos; San Segundo Acosta, Pablo
    Uno de los principales problemas con los que se enfrentan los profesores de Biología es el escaso interés que despierta esta asignatura en los estudiantes universitarios de algunas disciplinas -como química, física, arquitectura o ingeniería- al considerarla una asignatura difícil de aprender y de poca utilidad. Así, motivación, comprensión e interdisciplinaridad son tres retos a los que se enfrenta el profesorado de la asignatura de Biología del Grado en Química. El objetivo general del proyecto ha sido contribuir a la mejora de la enseñanza/aprendizaje de la asignatura de Biología. Para ello el profesorado ha implementado en el aula, y de forma combinada, dos metodologías: La "Biomímesis" y "La enseñanza/aprendizaje basada en Proyectos", que han demostrado ser herramientas muy útiles en el aula, para despertar la motivación y el interés de los estudiantes por el aprendizaje de Biología. Por un lado, al implicarles en su propio proceso de aprendizaje, los estudiantes han tenido un papel protagonista al ser los responsables del desarrollo de proyectos interesantes y retadores, elegidos por ellos mismos. Por otro lado, les ha permitido ver la aplicación o transferencia sostenible de lo aprendido en el aula a la sociedad y el medio ambiente. A estas dos metodologías didácticas se ha unido una tercera: Los Museos de Ciencia Naturales como espacios de enseñanza/aprendizaje de ciencia por investigación. Un total de 2 grupos de teoría han participado en esta experiencia docente. Los trabajos realizados han sido expuestos a otros estudiantes (en el aula) y al público general (en el Museo Nacional de Ciencias Naturales).
  • Publication
    Ash pollen immunoproteomics: Identification, immunologic characterization, and sequencing of 6 new allergens
    (Elsevier, 2014) Mas García, Salvador; Torres Pardo, María; Garrido-Arandia, María; Salamanca, Guillermo; Castro, Lourdes; Barral, Patricia; Purohit, Ashok; Pauli, Gabrielle; Rodríguez García, Rosalía; Batanero Cremades, Eva; Barderas Manchado, Rodrigo; Villalba Díaz, Mayte
    Immunoproteomics, 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.
  • Publication
    Allium porrum Extract Decreases Effector Cell Degranulation and Modulates Airway Epithelial Cell Function
    (MDPI, 2019-06-08) Benedé, Sara; Gradillas, Ana; Villalba Díaz, Mayte; Batanero Cremades, Eva
    Allium 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.
  • Publication
    Química Inspirada por la Naturaleza: Lecciones en el Museo Nacional de Ciencias Naturales II
    (2018-06-04) Batanero Cremades, Eva; Yélamos López, Belén; Fernández Fernández, María Inmaculada; López García-Gallo, Pilar; López-Rodríguez, Juan Carlos; San Segundo Acosta, Pablo; Rivera de Torre, Esperanza; Liñan Vargas, Katia Estefanía
    El proyecto Innova-Docencia 70 es una continuación del proyecto Innova-Docencia 30 que se desarrolló durante el curso académico 2016-2017 y su objetivo principal es implementar una nueva estrategia de enseñanza/aprendizaje que motive a los estudiantes hacia el estudio de Biología, favorezca la comprensión de los contenidos y les permita conectar los nuevos conocimientos adquiridos con el mundo real que les rodea. Motivación, comprensión e interdisciplinaridad son tres grandes retos a los que se enfrenta el profesorado de la asignatura de Biología del Grado en Química, que se imparte durante el primer cuatrimestre y que es de carácter obligatorio para los estudiantes de primero (6 ECTS), dado el escaso interés que despierta esta asignatura en los estudiantes de esta disciplina (además de otras, como física, arquitectura etc.), al considerarla una asignatura difícil de aprender y de poca utilidad. Esta estrategia docente tiene como eje metodológico principal la Biomímesis, pero a su vez se apoya en otras dos metodologías: (1) La enseñanza/aprendizaje basada en Proyectos y (2) Los Museos como espacios de enseñanza/aprendizaje. Todas ellas comparten un mismo principio: motivar al estudiante a aprender, implicándole en su propio proceso de aprendizaje. El estudiante ha de planificar, desarrollar y evaluar un proyecto (de carácter interdisciplinar) que le interesa, al ser elegido por él, y que le permite conectar lo académico con la vida.
  • Publication
    Surface activity as a crucial factor of the biological actions of Ole e1, the main aeroallergen of olive tree (Olea europaea) pollen
    (American Chemical Society, 2016-10-25) López-Rodríguez, Juan C.; Barderas Manchado, Rodrigo; Echaide Torreguitar, Mercedes; Pérez-Gil, Jesús; Villalba, Mayte; Batanero Cremades, Eva; Cruz Rodríguez, Antonio
    Aeroallergens are airborne substancesmainly proteinscapable of triggering Th2-immune responses in respiratory allergies. They enter into the body through the upper airways, reaching the mucosa afterward. Mucosae lining at the luminal side consists of an epithelial barrier completely covered by mucus and pulmonary surfactant. Both pulmonary surfactant and plasma membrane of the epithelial cells represent two physiological phospholipid-based barriers where allergens first impact before triggering their biological effects. The interaction of allergens with lipids at relevant physiological surfaces could promote structural changes on the molecule, resulting on a potential modification of its allergenic properties. In this work, we have first described the surface and phospholipid interaction capabilities of the clinically relevant aeroallergen Ole e 1, the main allergen of olive tree pollen. By using epifluorescence microscopy of Langmuir transferred films, we observed that lipid-packed ordered domains may function as a preferential location for allergen to accumulate at the air−liquid interface, an effect that is abolished in the presence of cholestenone. The possible implications of phospholipid-interfacial effects in the modification of allergen structural and functional properties will be discussed
  • Publication
    Human glutathione-S-transferase pi potentiates the cysteine-protease activity of the Der p 1 allergen from house dust mite through a cysteine redox mechanism
    (2019-09) López Rodríguez, Juan Carlos; Manosalva, Juliana; Cabrera-García, J. Daniel; Escribese, María M.; Villalba, Mayte; Barber, Domingo; Martínez Ruiz, Antonio; Batanero Cremades, Eva
    Environmental proteases have been widely associated to the pathogenesis of allergic disorders. Der p 1, a cysteine-protease from house dust mite (HDM) Dermatophagoides pteronyssinus, constitutes one of the most clinically relevant indoor aeroallergens worldwide. Der p 1 protease activity depends on the redox status of its catalytic cysteine residue, which has to be in the reduced state to be active. So far, it is unknown whether Der p 1-protease activity could be regulated by host redox microenvironment once it reaches the lung epithelial lining fluid in addition to endogenous mite components. In this sense, Glutathione-S-transferase pi (GSTpi), an enzyme traditionally linked to phase II detoxification, is highly expressed in human lung epithelial cells, which represent the first line of defence against aeroallergens. Moreover, GSTpi is a generalist catalyst of protein S-glutathionylation reactions, and some polymorphic variants of this enzyme has been associated to the development of allergic asthma. Here, we showed that human GSTpi increased the cysteine-protease activity of Der p 1, while GSTmu (the isoenzyme produced by the mite) did not alter it. GSTpi induces the reduction of Cys residues in Der p 1, probably by rearranging its disulphide bridges. Furthermore, GSTpi was detected in the apical medium collected from human bronchial epithelial cell cultures, and more interesting, it increased cysteine-protease activity of Der p 1. Our findings support the role of human GSTpi from airways in modulating of Der p 1 cysteineprotease activity, which may have important clinical implications for immune response to this aeroallergen in genetically susceptible individuals.