Person:
López Rodríguez, Juan Carlos

First Name

Juan Carlos

Last Name

López Rodríguez

URI

https://hdl.handle.net/20.500.14352/86792

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Ciencias Químicas

Department

Bioquímica y Biología Molecular

Area

Bioquímica y Biología Molecular

Identifiers

Full item page

Search Results

Now showing 1 - 9 of 9

Der p 1‑based immunotoxin as potential tool for the treatment of dust mite respiratory allergy
(Scientific Reports, 2020) Lázaro‑Gorines, Rodrigo; López Rodríguez, Juan Carlos; Benedé Pérez, Sara; González, Miguel; Mayorga, Cristobalina; Vogel, Lothar; Martínez Del Pozo, Álvaro; Lacadena García-Gallo, Francisco Javier; Villalba Díaz, María Teresa
Immunotoxins appear as promising therapeutic molecules, alternative to allergen-specifcimmunotherapy. In this work, we achieved the development of a protein chimera able to promote specifc cell death on efector cells involved in the allergic reaction. Der p 1 allergen was chosen as cell-targeting domain and the powerful ribotoxin α-sarcin as the toxic moiety. The resultant construction, named proDerp1αS, was produced and purifed from the yeast Pichia pastoris. Der p 1-protease activity and α-sarcin ribonucleolytic action were efectively conserved in proDerp1αS. Immunotoxin impact was assayed by using efector cells sensitized with house dust mite-allergic sera. Cell degranulation and death, triggered by proDerp1αS, was exclusively observed on Der p 1 sera sensitized-humRBL-2H3 cells, but not when treated with non-allergic sera. Most notably, equivalent IgE-binding and degranulation were observed with both proDerp1αS construct and native Der p 1 when using purifed basophils from sensitized patients. However, proDerp1αS did not cause any cytotoxic efect on these cells, apparently due to its lack of internalization after their surface IgEbinding, showing the complex in vivo panorama governing allergic reactions. In conclusion, herein we present proDerp1αS as a proof of concept for a potential and alternative new designs of therapeutic tools for allergies. Development of new, and more specifc, second-generation of immunotoxins following proDerp1αS, is further discussed
Estudio del papel del epitelio pulmonar en la alergia: el polen de olivo como modelo experimental
(2019) López Rodríguez, Juan Carlos; Batanero Cremades, Eva; Villalba Díaz, María Teresa
La alergia respiratoria es uno de los problemas de salud pública con mayor impacto social a nivel mundial, cuya prevalencia ha aumentado de forma alarmante durante los últimos años. Su etiología es compleja e implica la interacción de factores genéticos, factores ambientales y el estilo de vida de los países desarrollados. Este desorden clínico se caracteriza por un proceso inflamatorio de las vías respiratorias, dominado poruna respuesta inmune en la que participan linfocitos tipo T-colaboradores 2 (Th2) y la presencia de elevados niveles de inmunoglobulina E específica en el suero frente a sustancias ambientales, denominadas aeroalérgenos, que normalmente son inocuas para la mayoría de los individuos. Cada vez hay más evidencias del importante papel del epitelio respiratorio como integrador y regulador de la respuesta alérgica..
Immunologic responses to the major allergen of Olea Europaea in local and systemic allergic rhinitis subjects
(Clinical and Translational Allergy, 2015) Campo, Paloma; Villalba Díaz, Mayte; Barrionuevo, Esther; Rondón, Carmen; Galindo, Luisa; Rodríguez, María José; López Rodríguez, Juan Carlos; Torres, María José; Blanca, Miguel; Mayorga, Cristobalina
Evaluate: the in vivo and in vitro responses to nOle e 1 in allergic rhinitis (AR) and local allergic rhinitis (LAR) patients sensitized to olive tree pollen (OL) confirmed by nasal allergen provocation test (NAPT). Methods: Twelve subjects with AR, 12 with LAR, and 12 subjects as control group (CG) were selected. Skin testing and NAPT with nOle e 1 were performed. ECP and tryptase were measured in nasal lavages before and after NAPT. Serum IgE to OL allergens were measured by ELISA. Basophil activation tests (BAT) with OL and nOle e 1 and dendritic cell maturation/proliferation studies were carried out. Results: All AR (12/12) and 10/12 (83%) of LAR had a +NAPT to nOle e 1. ECP levels in nasal lavages were significantly increased after NAPT in both AR and LAR compared with CG at 15 minutes (p<0.05). Serum IgE was positive only in AR. All AR had +BAT responses to OL and 10/12 to nOle e 1 (83%); 8/12 LAR (66.6%) had a +BAT with OL and 4/12 (33%) to nOle e 1, with only one subject of the control group with a +BAT to both OL and nOle e 1 (8%). Dendritic cell proliferation to nOle e 1 was increased in AR compared to LAR and CG (p=0.019 and p=0.001 respectively).Conclusion: Both AR and LAR had a similar in vivo response to nOle e 1 with release of inflammatory mediators. Specific basophil activation with OL and nOle e 1 was observed in LAR confirming previous data obtained with dust mites.
Der p 1 based immunotoxin as potential tool for the treatment of dust mite respiratory allergy
(Scientific reports, 2020) Lázaro Gorines, Rodrigo; López Rodríguez, Juan Carlos; Benedé, Sara; González, Miguel; Mayorga, Cristobalina; Vogel, Lothar; Martínez del Pozo, Álvaro; Lacadena, Javier; Villalba, Mayte
Immunotoxins appear as promising therapeutic molecules, alternative to allergen-specifcimmunotherapy. In this work, we achieved the development of a protein chimera able to promote specifc cell death on efector cells involved in the allergic reaction. Der p 1 allergen was chosen as cell-targeting domain and the powerful ribotoxin α-sarcin as the toxic moiety. The resultant construction, named proDerp1αS, was produced and purifed from the yeast Pichia pastoris. Der p 1-protease activity and α-sarcin ribonucleolytic action were efectively conserved in proDerp1αS. Immunotoxin impact was assayed by using efector cells sensitized with house dust mite-allergic sera. Cell degranulation and death, triggered by proDerp1αS, was exclusively observed on Der p 1 sera sensitized-humRBL-2H3 cells, but not when treated with non-allergic sera. Most notably, equivalent IgE-binding and degranulation were observed with both proDerp1αS construct and native Der p 1 when using purifed basophils from sensitized patients. However, proDerp1αS did not cause any cytotoxic efect on these cells, apparently due to its lack of internalization after their surface IgEbinding, showing the complex in vivo panorama governing allergic reactions. In conclusion, herein we present proDerp1αS as a proof of concept for a potential and alternative new designs of therapeutic tools for allergies. Development of new, and more specifc, second-generation of immunotoxins following proDerp1αS, is further discussed.
Biophysical and biological impact on the structure and IgE-binding of the interaction of the olive pollen allergen Ole e 7 with lipids
(Biochimica et Biophysica Acta (BBA) - Biomembranes, 2020) Oeo-Santos, Carmen; López Rodríguez, Juan Carlos; García Mouton, Cristina; San Segundo Acosta, Pablo; Jurado, Aurora; Moreno-Aguilar, Carmen; García Álvarez, María Begoña; Pérez Gil, Jesús; Villalba Díaz, María Teresa; Barderas Manchado, Rodrigo; Cruz Rodríguez, Antonio
Ole e 7 allergen from Olea europaea pollen possesses a major clinical relevance because it produces severe symptoms, such as anaphylaxis, in allergic patients exposed to high olive pollen counts. Ole e 7 is a non-specific lipid transfer protein (nsLTP) characterized by the presence of a tunnel-like hydrophobic cavity, which may be suitable for hosting and, thus, transporting lipids -as it has been described for other nsLTPs-. The identification of the primary amino acid sequence of Ole e 7, and its production as a recombinant allergen, allowed characterizing its lipid-binding properties and its effect at air-liquid interfaces. Fluorescence and interferometry experiments were performed using different phospholipid molecular species and free fatty acids to analyse the lipid-binding ability and specificity of the allergen. Molecular modelling of the allergen was used to determine the potential regions involved in lipid interaction. Changes in Ole e 7 structure after lipid interaction were analysed by circular dichroism. Changes in the IgE binding upon ligand interaction were determined by ELISA. Wilhelmy balance measurements and fluorescence surfactant adsorption tests were performed to analyse the surface activity of the allergen. Using these different approaches, we have demonstrated the ability of Ole e 7 to interact and bind to a wide range of lipids, especially negatively charged phospholipids and oleic acid. We have also identified the protein structural regions and the residues potentially involved in that interaction, suggesting how lipid-protein interactions could define the behaviour of the allergen once inhaled at the airways.
Project number: 30
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).
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
(Redox Biology, 2019) 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.
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 Javier
The “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.
Molecular dissection of the membrane aggregation mechanisms induced by monomeric annexin A2
(Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 2018) López Rodríguez, Juan Carlos; Martínez-Carmona, Francisco J.; Rodríguez Crespo, José Ignacio; Lizarbe Iracheta, María Antonia; Turnay Abad, Francisco Javier
Annexins are a multigene family of proteins involved in aggregation and fusion processes of biological membranes. One of its best-known members is annexin A2 (or p36), capable of binding to acidic phospholipids in a calcium-dependent manner, as occurs with other members of the same family. In its heterotetrameric form, especially with protein S100A10 (p11), annexin A2 has been involved as a determinant factor in innumerable biological processes like tumor development or anticoagulation. However, the subcellular coexistence of different pools of the protein, in which the monomeric form of annexin A2 is growing in functional relevance, is to date poorly described. In this work we present an exhaustive structural and functional characterization of monomeric human annexin A2 by using different recombinant mutants. The important role of the amphipathic N-terminal α-helix in membrane binding and aggregation has been analyzed. We have also studied the potential implication of lateral "antiparallel" protein dimers in membrane aggregation. In contrast to what was previously suggested, formation of these dimers negatively regulate aggregation. We have also confirmed the essential role of three lysine residues located in the convex surface of the molecule in calcium-free and calcium-dependent membrane binding and aggregation. Finally, we propose models for annexin A2-mediated vesicle aggregation mechanisms.