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1. The TNF-like weak inducer of the apoptosis/fibroblast growth factor–inducible molecule 14 axis mediates histamine and platelet-activating factor–induced subcutaneous vascular leakage and anaphylactic shock

Author

Mendez-Barbero, Nerea, Yuste-Montalvo, Alma, Nuñez-Borque, Emilio, Jensen, Bettina M., Gutiérrez-Muñoz, Carmen, Tome-Amat, Jaime, Garrido-Arandia, María, Díaz-Perales, Araceli, Ballesteros-Martinez, Contanza, Laguna, Jose Julio, Beitia, J.M., Poulsen, Lars K., Cuesta-Herranz, Javier, Blanco-Colio, Luis Miguel, and Esteban, Vanesa

Published
2020
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2. Critical features of an in vitro intestinal absorption model to study the first key aspects underlying food allergen sensitization

Author

l’Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (France), Centro de Biotecnología y Genómica de Plantas (España), Universidad Politécnica de Madrid, CSIC-UAM - Instituto de Investigación en Ciencias de la Alimentación (CIAL), Fundação para a Ciência e a Tecnologia (Portugal), Fondation pour la Recherche Médicale, Ministerio de Economía, Industria y Competitividad (España), Agencia Estatal de Investigación (España), Ministerio de Ciencia e Innovación (España), Région des Pays de la Loire, Ministerio de Ciencia, Innovación y Universidades (España), Comissão de Coordenação e Desenvolvimento Regional do Norte (Portugal), European Commission, Ministry of Agriculture, Nature and Food Quality (The Netherlands), Dijk, Wieneke [0000-0003-1676-7751], Villa, Caterina [0000-0002-9471-3612], Benedé, Sara [0000-0002-9288-9438], Vassilopoulou, Emilia [0000-0002-2665-5908], Mafra, Isabel [0000-0001-5311-8895], Garrido-Arandía, María [0000-0001-6114-5754], Martínez Blanco, Mónica [0000-0003-1153-7957], Bouchaud, Gregory [0000-0002-3794-7287], Hoppenbrouwers, Tamara [0000-0002-1764-9100], Bavaro, Simona Lucia [0000-0002-5820-8088], Giblin, Linda [0000-0002-9354-3121], Knipping, Karen [0000-0002-3700-9047], Costa, Joana [0000-0002-8229-2902], Bastiaan-Net, Shanna [0000-0001-8922-7305], Dijk, Wieneke, Villa, Caterina, Benedé, Sara, Vassilopoulou, Emilia, Mafra, Isabel, Garrido-Arandía, María, Martínez-Blanco, Mónica, Bouchaud, Grégory, Hoppenbrouwers, Tamara, Bavaro, Simona L., Giblin, Linda, Knipping, Karen, Castro Reigía, Ana María, Delgado, Susana, Costa, Joana, Bastiaan-Net, Shanna, l’Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (France), Centro de Biotecnología y Genómica de Plantas (España), Universidad Politécnica de Madrid, CSIC-UAM - Instituto de Investigación en Ciencias de la Alimentación (CIAL), Fundação para a Ciência e a Tecnologia (Portugal), Fondation pour la Recherche Médicale, Ministerio de Economía, Industria y Competitividad (España), Agencia Estatal de Investigación (España), Ministerio de Ciencia e Innovación (España), Région des Pays de la Loire, Ministerio de Ciencia, Innovación y Universidades (España), Comissão de Coordenação e Desenvolvimento Regional do Norte (Portugal), European Commission, Ministry of Agriculture, Nature and Food Quality (The Netherlands), Dijk, Wieneke [0000-0003-1676-7751], Villa, Caterina [0000-0002-9471-3612], Benedé, Sara [0000-0002-9288-9438], Vassilopoulou, Emilia [0000-0002-2665-5908], Mafra, Isabel [0000-0001-5311-8895], Garrido-Arandía, María [0000-0001-6114-5754], Martínez Blanco, Mónica [0000-0003-1153-7957], Bouchaud, Gregory [0000-0002-3794-7287], Hoppenbrouwers, Tamara [0000-0002-1764-9100], Bavaro, Simona Lucia [0000-0002-5820-8088], Giblin, Linda [0000-0002-9354-3121], Knipping, Karen [0000-0002-3700-9047], Costa, Joana [0000-0002-8229-2902], Bastiaan-Net, Shanna [0000-0001-8922-7305], Dijk, Wieneke, Villa, Caterina, Benedé, Sara, Vassilopoulou, Emilia, Mafra, Isabel, Garrido-Arandía, María, Martínez-Blanco, Mónica, Bouchaud, Grégory, Hoppenbrouwers, Tamara, Bavaro, Simona L., Giblin, Linda, Knipping, Karen, Castro Reigía, Ana María, Delgado, Susana, Costa, Joana, and Bastiaan-Net, Shanna

Abstract

New types of protein sources will enter our diet in a near future, reinforcing the need for a straightforward in vitro (cell-based) screening model to test and predict the safety of these novel proteins, in particular their potential risk for de novo allergic sensitization. The Adverse Outcome Pathway (AOP) for allergen sensitization describes the current knowledge of key events underlying the complex cellular interactions that proceed allergic food sensitization. Currently, there is no consensus on the in vitro model to study the intestinal translocation of proteins as well as the epithelial activation, which comprise the first molecular initiation events (ME1-3) and the first key event of the AOP, respectively. As members of INFOGEST, we have highlighted several critical features that should be considered for any proposed in vitro model to study epithelial protein transport in the context of allergic sensitization. In addition, we defined which intestinal cell types are indispensable in a consensus model of the first steps of the AOP, and which cell types are optional or desired when there is the possibility to create a more complex cell model. A model of these first key aspects of the AOP can be used to study the gut epithelial translocation behavior of known hypo- and hyperallergens, juxtaposed to the transport behavior of novel proteins as a first screen for risk management of dietary proteins. Indeed, this disquisition forms a basis for the development of a future consensus model of the allergic sensitization cascade, comprising also the other key events (KE2-5).

Published
2023

3. Corrigendum: Suitability of potyviral recombinant virus-like particles bearing a complete food allergen for immunotherapy vaccines(Front. Immunol., (2023), 13, (986823), 10.3389/fimmu.2022.986823)

Author

Garrido-Arandía, María [0000-0001-6114-5754], Ponz Ascaso, Fernando [0000-0003-0344-4363], Díaz-Perales, Araceli [0000-0002-1093-3627], Pazos-Castro, Diego, Margain, Clémence, Gonzalez-Klein, Zulema, Amores-Borge, Marina, Yuste-Calvo, C., Garrido-Arandía, María, Zurita, Lucía, Esteban, Vanesa, Tome-Amat, Jaime, Díaz-Perales, Araceli, Ponz Ascaso, Fernando, Garrido-Arandía, María [0000-0001-6114-5754], Ponz Ascaso, Fernando [0000-0003-0344-4363], Díaz-Perales, Araceli [0000-0002-1093-3627], Pazos-Castro, Diego, Margain, Clémence, Gonzalez-Klein, Zulema, Amores-Borge, Marina, Yuste-Calvo, C., Garrido-Arandía, María, Zurita, Lucía, Esteban, Vanesa, Tome-Amat, Jaime, Díaz-Perales, Araceli, and Ponz Ascaso, Fernando

Abstract

In the published article, there was an error in the author list, and author Marina Amores-Borge was erroneously excluded. The corrected author list appears below. “Diego Pazos-Castro, Clémence Margain, Zulema Gonzalez-Klein, Marina Amores- Borge, Carmen Yuste-Calvo, Maria Garrido-Arandia, Lucia Zurita, Vanesa Esteban, Jaime Tome-Amat, Araceli Diaz-Perales, Fernando Ponz”In the published article, there was an error. The missing author was not included in the Author Contributions section. A correction has been made to Author contributions. This sentence previously stated: “DP-C: Conceptualization, investigation, writing original draft. CM: Investigation. ZG-K: Conceptualization, investigation, writing review. CY-C: Investigation. MG-A: Investigation, writing review. LZ: Investigation. VE: Writing review, resources, funding acquisition. JT-A: Conceptualization, investigation, writing original draft. AD-P: Conceptualization, writing original draft, resources, funding acquisition. FP: Conceptualization, writing review,resources, funding acquisition. All authors contributed to the article and approved the submitted version.” The corrected sentence appears below: “DP-C: Conceptualization, investigation, writing original draft. CM: Investigation. ZG-K: Conceptualization, investigation, writing review. MA-B: Investigation. CY-C: Investigation. MG-A: Investigation, writing review. LZ: Investigation. VE: Writing review, resources, funding acquisition. JT-A: Conceptualization, investigation, writing original draft. AD-P: Conceptualization, writing original draft, resources, funding acquisition. FP: Conceptualization, writing review, resources, funding acquisition. All authors contributed to the article and approved the submitted version.” The authors apologize for these errors and state that they do not change the scientific conclusions of the article in any way. The original article has been updated.

Published
2023

4. Statistical Study of Low-Intensity Single-Molecule Recognition Events Using DeepTipTM Probes: Application to the Pru p 3-Phytosphingosine System

Author

Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Comunidad de Madrid, European Commission, Universidad Politécnica de Madrid, Banco Santander, Daza, Rafael [0000-0001-7883-648X], Garrido-Arandía, María [0000-0001-6114-5754], Corregidor-Ortiz, Daniel [0000-0002-8881-8880], Colchero, Luis [0000-0002-9673-0061], Tabraue-Rubio, Raquel [0000-0001-7974-689X], Elices, Manuel [0000-0002-8204-1580], Guinea, Gustavo V. [0000-0002-4326-3746], Díaz-Perales, Araceli [0000-0002-1093-3627], Pérez-Rigueiro, José [0000-0001-8298-8398], Daza, Rafael, Garrido-Arandía, María, Corregidor-Ortiz, Daniel, Pérez, Carla Isabel, Colchero, Luis, Tabraue-Rubio, Raquel, Elices, Manuel, Guinea, Gustavo V., Díaz-Perales, Araceli, Pérez-Rigueiro, José, Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Comunidad de Madrid, European Commission, Universidad Politécnica de Madrid, Banco Santander, Daza, Rafael [0000-0001-7883-648X], Garrido-Arandía, María [0000-0001-6114-5754], Corregidor-Ortiz, Daniel [0000-0002-8881-8880], Colchero, Luis [0000-0002-9673-0061], Tabraue-Rubio, Raquel [0000-0001-7974-689X], Elices, Manuel [0000-0002-8204-1580], Guinea, Gustavo V. [0000-0002-4326-3746], Díaz-Perales, Araceli [0000-0002-1093-3627], Pérez-Rigueiro, José [0000-0001-8298-8398], Daza, Rafael, Garrido-Arandía, María, Corregidor-Ortiz, Daniel, Pérez, Carla Isabel, Colchero, Luis, Tabraue-Rubio, Raquel, Elices, Manuel, Guinea, Gustavo V., Díaz-Perales, Araceli, and Pérez-Rigueiro, José

Abstract

The interaction between the plant lipid transfer protein Pru p 3 and phytosphingosine was assessed using an atomic force microscope. Phytosphingosine was covalently immobilized on DeepTipTM probes and Pru p 3 on MicroDeckTM functionalized substrates. Single-molecular interaction events between both molecules were retrieved and classified and the distribution for each one of the identified types was calculated. A success rate of over 70% was found by comparing the number of specific Pru p 3-phytosphingosine interaction events with the total number of recorded curves. The analysis of the distribution established among the various types of curves was further pursued to distinguish between those curves that can mainly be used for assessing the recognition between phytosphingosine (sensor molecule) and Pru p 3 (target molecule) in the context of affinity atomic force microscopy, and those that entail details of the interaction and might be employed in the context of force spectroscopy. The successful application of these functionalized probes and substrates to the characterization of the low-intensity hydrophobic interaction characteristic of this system is a clear indication of the potential of exploiting this approach with an extremely wide range of different biological molecules of interest. The possibility of characterizing molecular assembly events with single-molecule resolution offers an advantageous procedure to plough into the field of molecular biomimetics.

Published
2023

5. Natural variation in Avr3D1 from Zymoseptoria sp. contributes to quantitative gene-for-gene resistance and to host specificity

Author

Swiss National Science Foundation, Ministerio de Ciencia e Innovación (España), Ministerio de Ciencia, Innovación y Universidades (España), Meile, Lukas [0000-0002-2680-3309], Garrido-Arandía, María [0000-0001-6114-5754], Bernasconi, Zoe [0000-0002-1216-6791], Bernasconi, Alessio [0000-0003-3833], Alassimone, Julien [0000-0002-8118-2605], McDonald, Bruce A. [0000-0002-5332-2172], Sánchez-Vallet, Andrea [0000-0002-3668-9503], Meile, Lukas, Garrido-Arandía, María, Bernasconi, Zoe, Peter, Jules, Schneller, Alissa, Bernasconi, Alessio, Alassimone, Julien, McDonald, Bruce A., Sánchez-Vallet, Andrea, Swiss National Science Foundation, Ministerio de Ciencia e Innovación (España), Ministerio de Ciencia, Innovación y Universidades (España), Meile, Lukas [0000-0002-2680-3309], Garrido-Arandía, María [0000-0001-6114-5754], Bernasconi, Zoe [0000-0002-1216-6791], Bernasconi, Alessio [0000-0003-3833], Alassimone, Julien [0000-0002-8118-2605], McDonald, Bruce A. [0000-0002-5332-2172], Sánchez-Vallet, Andrea [0000-0002-3668-9503], Meile, Lukas, Garrido-Arandía, María, Bernasconi, Zoe, Peter, Jules, Schneller, Alissa, Bernasconi, Alessio, Alassimone, Julien, McDonald, Bruce A., and Sánchez-Vallet, Andrea

Abstract

Successful host colonization by plant pathogens requires the circumvention of host defense responses, frequently through sequence modifications in secreted pathogen proteins known as avirulence factors (Avrs). Although Avr sequences are often polymorphic, the contribution of these polymorphisms to virulence diversity in natural pathogen populations remains largely unexplored. We used molecular genetic tools to determine how natural sequence polymorphisms of the avirulence factor Avr3D1 in the wheat pathogen Zymoseptoria tritici contributed to adaptive changes in virulence. We showed that there is a continuous distribution in the magnitude of resistance triggered by different Avr3D1 isoforms and demonstrated that natural variation in an Avr gene can lead to a quantitative resistance phenotype. We further showed that homologues of Avr3D1 in two nonpathogenic sister species of Z. tritici are recognized by some wheat cultivars, suggesting that Avr-R gene-for-gene interactions can contribute to nonhost resistance. We suggest that the mechanisms underlying host range, qualitative resistance, and quantitative resistance are not exclusive.

Published
2022

6. Suitability of potyviral recombinant virus-like particles bearing a complete food allergen for immunotherapy vaccines

Author

Comunidad de Madrid, European Commission, Ministerio de Ciencia e Innovación (España), Instituto de Salud Carlos III, Universidad Politécnica de Madrid, Pazos-Castro, Diego [0000-0002-0686-5131], Gonzalez-Klein, Zulema [0000-0003-3762-3781], Garrido-Arandía, María [0000-0001-6114-5754], Zurita, Lucía [0000-0001-5523-1832], Tome-Amat, Jaime [0000-0003-4442-3649], Díaz-Perales, Araceli [0000-0002-1093-3627], Ponz Ascaso, Fernando [0000-0003-0344-4363], Pazos-Castro, Diego, Margain, C., Gonzalez-Klein, Zulema, Amores-Borge, Marina, Yuste-Calvo, C., Garrido-Arandía, María, Zurita, Lucía, Esteban, Vanesa, Tome-Amat, Jaime, Díaz-Perales, Araceli, Ponz Ascaso, Fernando, Comunidad de Madrid, European Commission, Ministerio de Ciencia e Innovación (España), Instituto de Salud Carlos III, Universidad Politécnica de Madrid, Pazos-Castro, Diego [0000-0002-0686-5131], Gonzalez-Klein, Zulema [0000-0003-3762-3781], Garrido-Arandía, María [0000-0001-6114-5754], Zurita, Lucía [0000-0001-5523-1832], Tome-Amat, Jaime [0000-0003-4442-3649], Díaz-Perales, Araceli [0000-0002-1093-3627], Ponz Ascaso, Fernando [0000-0003-0344-4363], Pazos-Castro, Diego, Margain, C., Gonzalez-Klein, Zulema, Amores-Borge, Marina, Yuste-Calvo, C., Garrido-Arandía, María, Zurita, Lucía, Esteban, Vanesa, Tome-Amat, Jaime, Díaz-Perales, Araceli, and Ponz Ascaso, Fernando

Abstract

Virus-like particles (VLPs) have been gaining attention as potential platforms for delivery of cargos in nanomedicine. Although animal viruses are largely selected due to their immunostimulatory capacities, VLPs from plant viruses constitute a promising alternative to be considered. VLPs derived from Turnip mosaic virus (TuMV) have proven to present a tridimensional structure suited to display molecules of interest on their surface, making them interesting tools to be studied in theragnostic strategies. Here, we study their potential in the treatment of food allergy by genetically coupling TuMV-derived VLPs to Pru p 3, one of the most dominant allergens in Mediterranean climates. VLPs-Pru p 3 were generated by cloning a synthetic gene encoding the TuMV coat protein and Pru p 3, separated by a linker, into a transient high-expression vector, followed by agroinfiltration in Nicotiana benthamiana plants. The generated fusion protein self-assembled in planta to form the VLPs, which were purified by exclusion chromatography. Their elongated morphology was confirmed by electron microscopy and their size (~400 nm), and monodispersity was confirmed by dynamic light scattering. Initial in vitro characterization confirmed that they were able to induce proliferation of human immune cells. This proliferative capability was enhanced when coupled with the natural lipid ligand of Pru p 3. The resultant formulation, called VLP-Complex, was also able to be transported by intestinal epithelial cells, without affecting the monolayer integrity. In light of all these results, VLP-Complex was furtherly tested in a mouse model of food allergy. Sublingual administration of VLP-Complex could effectively reduce some serological markers associated with allergic responses in mice, such as anti Pru p 3 sIgE and sIgG2a. Noteworthy, no associated macroscopic, nephritic, or hepatic toxicity was detected, as assessed by weight, blood urea nitrogen (BUN) and galectin-3 analyses, respectively.

Published
2022

7. Allergen Profile of London Plane Tree Pollen: Clinical and Molecular Pattern in Central Spain

Author

Instituto de Salud Carlos III, European Commission, Nuñez-Borque, E. [0000-0003-2928-410X], Gómez-Cardeñosa, A. [0000-0001-5077-2509], Garrido-Arandía, María [0000-0001-6114-5754], Nuñez-Borque, Emilio, Betancor, Diana, Fernandez-Bravo, Sergio, Gómez-Cardeñosa, A., Esteban, Vanesa, Garrido-Arandía, María, de Las Heras, M., Pastor-Vargas, C., Cuesta, Javier, Instituto de Salud Carlos III, European Commission, Nuñez-Borque, E. [0000-0003-2928-410X], Gómez-Cardeñosa, A. [0000-0001-5077-2509], Garrido-Arandía, María [0000-0001-6114-5754], Nuñez-Borque, Emilio, Betancor, Diana, Fernandez-Bravo, Sergio, Gómez-Cardeñosa, A., Esteban, Vanesa, Garrido-Arandía, María, de Las Heras, M., Pastor-Vargas, C., and Cuesta, Javier

Abstract

Platanus acerifolia (London plane tree) is a deciduous tree of the Platanaceae family. Sensitization to this plant varies with geography. Madrid, located in central Spain, has one of the highest London plane tree pollen concentration levels on the Iberian Peninsula. We evaluated both the clinical characteristics and the molecular sensitization pattern of patients with allergy to London plane tree pollen in the region of Madrid.

Published
2022

8. Alt a 1 Promotes Allergic Asthma In Vivo Through TLR4-Alveolar Macrophages

Author

Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Universidad Politécnica de Madrid, Comunidad de Madrid, Instituto de Salud Carlos III, Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Pazos-Castro, Diego [0000-0002-0686-5131], Gonzalez-Klein, Zulema [0000-0003-3762-3781], Resuela-Gonzalez, Jose Luis [0000-0001-5443-0901], Fernandez-Bravo, Sergio [0000-0001-5291-0100], Palacio-Garcia, Lucia [0000-0003-2010-4686], Garrido-Arandía, María [0000-0001-6114-5754], Tome-Amat, Jaime [0000-0003-4442-3649], Díaz-Perales, Araceli [0000-0002-1093-3627], Hernandez-Ramirez, Guadalupe, Pazos-Castro, Diego, Gonzalez-Klein, Zulema, Resuela-Gonzalez, Jose Luis, Fernandez-Bravo, Sergio, Palacio-Garcia, Lucia, Esteban, Vanesa, Garrido-Arandía, María, Tome-Amat, Jaime, Díaz-Perales, Araceli, Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Universidad Politécnica de Madrid, Comunidad de Madrid, Instituto de Salud Carlos III, Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Pazos-Castro, Diego [0000-0002-0686-5131], Gonzalez-Klein, Zulema [0000-0003-3762-3781], Resuela-Gonzalez, Jose Luis [0000-0001-5443-0901], Fernandez-Bravo, Sergio [0000-0001-5291-0100], Palacio-Garcia, Lucia [0000-0003-2010-4686], Garrido-Arandía, María [0000-0001-6114-5754], Tome-Amat, Jaime [0000-0003-4442-3649], Díaz-Perales, Araceli [0000-0002-1093-3627], Hernandez-Ramirez, Guadalupe, Pazos-Castro, Diego, Gonzalez-Klein, Zulema, Resuela-Gonzalez, Jose Luis, Fernandez-Bravo, Sergio, Palacio-Garcia, Lucia, Esteban, Vanesa, Garrido-Arandía, María, Tome-Amat, Jaime, and Díaz-Perales, Araceli

Published
2022

9. NLRP3 priming due to skin damage precedes LTP allergic sensitization in a mouse model

Author

Ministerio de Economía y Competitividad (España), Instituto de Salud Carlos III, European Commission, Comunidad de Madrid, Universidad Politécnica de Madrid, Banco Santander, Pazos-Castro, Diego [0000-0002-0686-5131], Gonzalez-Klein, Zulema [0000-0003-3762-3781], Montalvo, Alma Yuste [0000-0002-4727-1284], Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Romero-Sahagun, Alejandro [0000-0002-9417-8293], Garrido-Arandía, María [0000-0001-6114-5754], Tome-Amat, Jaime [0000-0003-4442-3649], Díaz-Perales, Araceli [0000-0002-1093-3627], Pazos-Castro, Diego, Gonzalez-Klein, Zulema, Montalvo, Alma Yuste, Hernandez-Ramirez, Guadalupe, Romero-Sahagun, Alejandro, Esteban, Vanesa, Garrido-Arandía, María, Tome-Amat, Jaime, Díaz-Perales, Araceli, Ministerio de Economía y Competitividad (España), Instituto de Salud Carlos III, European Commission, Comunidad de Madrid, Universidad Politécnica de Madrid, Banco Santander, Pazos-Castro, Diego [0000-0002-0686-5131], Gonzalez-Klein, Zulema [0000-0003-3762-3781], Montalvo, Alma Yuste [0000-0002-4727-1284], Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Romero-Sahagun, Alejandro [0000-0002-9417-8293], Garrido-Arandía, María [0000-0001-6114-5754], Tome-Amat, Jaime [0000-0003-4442-3649], Díaz-Perales, Araceli [0000-0002-1093-3627], Pazos-Castro, Diego, Gonzalez-Klein, Zulema, Montalvo, Alma Yuste, Hernandez-Ramirez, Guadalupe, Romero-Sahagun, Alejandro, Esteban, Vanesa, Garrido-Arandía, María, Tome-Amat, Jaime, and Díaz-Perales, Araceli

Abstract

Allergic sensitization is initiated by protein and epithelia interaction, although the molecular mechanisms leading this encounter toward an allergic phenotype remain unknown. Here, we apply the two-hit hypothesis of inflammatory diseases to the study of food allergy sensitization. First, we studied the effects of long-term depilation in mice by analyzing samples at different time points. Several weeks of depilation were needed until clear immunological changes were evidenced, starting with upregulation of NLRP3 protein levels, which was followed by overexpression of Il1b and Il18 transcripts. Secondly, we assessed the effects of allergen addition (in this case, Pru p 3 in complex with its natural lipid ligand) over depilated skin. Systemic sensitization was evaluated by intraperitoneal provocation with Pru p 3 and measure of body temperature. Anaphylaxis was achieved, but only in mice sensitized with Prup3_complex and not treated with the NLRP3 inhibitor MCC950, thus demonstrating the importance of both hits (depilation + allergen addition) in the consecution of the allergic phenotype. In addition, allergen encounter (but not depilation) promoted skin remodeling, as well as CD45+ infiltration not only in the sensitized area (the skin), but across several mucosal tissues (skin, lungs, and gut), furtherly validating the systemization of the response. Finally, a low-scale study with human ILC2s is reported, where we demonstrate that Prup3_complex can induce their phenotype switch (↑CD86, ↑S1P1) when cultured in vitro, although more data is needed to understand the implications of these changes in food allergy development.

Published
2022

10. Plant non-specific lipid transfer proteins: An overview

Author

Ministère de l’Enseignement Supérieur et de la Recherche Scientifique (Tunisie), Comunidad de Madrid, Universidad Politécnica de Madrid, Banco Santander, European Commission, Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación (España), Gonzalez-Klein, Zulema [0000-0003-3762-3781], Pazos-Castro, Diego [0000-0002-0686-5131], Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Garrido-Arandía, María [0000-0001-6114-5754], Brini, Faical [0000-0002-8435-381X], Díaz-Perales, Araceli [0000-0002-1093-3627], Tome-Amat, Jaime [0000-0003-4442-3649], Missaoui, Khawla, Gonzalez-Klein, Zulema, Pazos-Castro, Diego, Hernandez-Ramirez, Guadalupe, Garrido-Arandía, María, Brini, Faiçal, Díaz-Perales, Araceli, Tome-Amat, Jaime, Ministère de l’Enseignement Supérieur et de la Recherche Scientifique (Tunisie), Comunidad de Madrid, Universidad Politécnica de Madrid, Banco Santander, European Commission, Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación (España), Gonzalez-Klein, Zulema [0000-0003-3762-3781], Pazos-Castro, Diego [0000-0002-0686-5131], Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Garrido-Arandía, María [0000-0001-6114-5754], Brini, Faical [0000-0002-8435-381X], Díaz-Perales, Araceli [0000-0002-1093-3627], Tome-Amat, Jaime [0000-0003-4442-3649], Missaoui, Khawla, Gonzalez-Klein, Zulema, Pazos-Castro, Diego, Hernandez-Ramirez, Guadalupe, Garrido-Arandía, María, Brini, Faiçal, Díaz-Perales, Araceli, and Tome-Amat, Jaime

Abstract

Plant non-specific lipid transfer proteins (nsLTPs) are usually defined as small, basic proteins, with a wide distribution in all orders of higher plants. Structurally, nsLTPs contain a conserved motif of eight cysteines, linked by four disulphide bonds, and a hydrophobic cavity in which the ligand is housed. This structure confers stability and enhances the ability to bind and transport a variety of hydrophobic molecules. Their highly conserved structural resemblance but low sequence identity reflects the wide variety of ligands they can carry, as well as the broad biological functions to which they are linked to, such as membrane stabilization, cell wall organization and signal transduction. In addition, they have also been described as essential in resistance to biotic and abiotic stresses, plant growth and development, seed development, and germination. Hence, there is growing interest in this family of proteins for their critical roles in plant development and for the many unresolved questions that need to be clarified, regarding their subcellular localization, transfer capacity, expression profile, biological function, and evolution.

Published
2022

11. Identification and molecular characterization of a novel non-specific lipid transfer protein (TdLTP2) from durum wheat

Author

Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Comunidad de Madrid, Ministère de l’Enseignement Supérieur et de la Recherche Scientifique (Tunisie), Gonzalez-Klein, Zulema [0000-0003-3762-3781], Jemli, Sonia [0000-0001-6265-581X], Garrido-Arandía, María [0000-0001-6114-5754], Díaz-Perales, Araceli [0000-0002-1093-3627], Tome-Amat, Jaime [0000-0003-4442-3649], Brini, Faiçal [0000-0002-8435-381X], Missaoui, Khawla, Gonzalez-Klein, Zulema, Jemli, Sonia, Garrido-Arandía, María, Díaz-Perales, Araceli, Tome-Amat, Jaime, Brini, Faiçal, Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Comunidad de Madrid, Ministère de l’Enseignement Supérieur et de la Recherche Scientifique (Tunisie), Gonzalez-Klein, Zulema [0000-0003-3762-3781], Jemli, Sonia [0000-0001-6265-581X], Garrido-Arandía, María [0000-0001-6114-5754], Díaz-Perales, Araceli [0000-0002-1093-3627], Tome-Amat, Jaime [0000-0003-4442-3649], Brini, Faiçal [0000-0002-8435-381X], Missaoui, Khawla, Gonzalez-Klein, Zulema, Jemli, Sonia, Garrido-Arandía, María, Díaz-Perales, Araceli, Tome-Amat, Jaime, and Brini, Faiçal

Abstract

Non-specific lipid transfer proteins (nsLTPs) are small, cysteine-rich proteins, a part of the pathogenesis-related protein family, and numerous of them act as positive regulators during plant disease resistance, growth, and reproduction. These proteins are involved also in the intracellular transfer of lipids, as well as in plant immune responses. Besides their differences in sequences, they show similar features in their structure. However, they show distinct lipid-binding specificities signifying their various biological roles that dictate further structural study. This study reports the identification, in silico characterization and purification of a novel member of the nsLTP2 protein family from durum wheat, TdLTP2. It was generated and purified using the combination of gel filtration chromatography and reverse-phase high-performance liquid chromatography (RP-HPLC). Its identity was detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry (MALDI-TOF). TdLTP2 had been expressed in different stress to detect its localization; therefore, fluor-immunolocalization studies accomplished this data. In this approach, to assess the allergenicity of TdLTP2, thirty patients with baker's asthma were enrolled and ELISA to detect the presence of specific IgE antibodies tested their sera. Moreover, the lipid-binding properties of TdLTP2 were examined in vitro and validated using a molecular docking study. In summary, our results demonstrate a new addition of member in plant nsLTPs family, TdLTP2, which can develop a better understanding about its biological functions and shed light on future applications.

Published
2022

12. Arabidopsis immune responses triggered by cellulose and mixed‐linked glucan oligosaccharides require a group of Leucine‐Rich Repeat‐Malectin Receptor Kinases

Author

Martín‐Dacal, Marina, primary, Fernández‐Calvo, Patricia, additional, Jiménez‐Sandoval, Pedro, additional, López, Gemma, additional, Garrido‐Arandía, María, additional, Rebaque, Diego, additional, del Hierro, Irene, additional, Berlanga, Diego José, additional, Torres, Miguel Ángel, additional, Kumar, Varun, additional, Mélida, Hugo, additional, Pacios, Luis F., additional, Santiago, Julia, additional, and Molina, Antonio, additional

Published
2022
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14. The Role of Sphingolipids in Allergic Disorders

Author

Ministerio de Economía y Competitividad (España), Agencia Estatal de Investigación (España), Instituto de Salud Carlos III, European Commission, Comunidad de Madrid, Díaz-Perales, Araceli [0000-0002-1093-3627], Escribese Maria M. [0000-0001-5057-5150], Garrido-Arandía, María [0000-0001-6114-5754], Obeso, David [0000-0001-7875-7327], Izquierdo-Alvarez, Elena [0000-0002-3355-2798], Tome-Amat, Jaime [0000-0003-4442-3649], Barber, Domingo F. [0000-0001-8824-5405], Díaz-Perales, Araceli, Escribese Maria M., Garrido-Arandía, María, Obeso, David, Izquierdo-Alvarez, Elena, Tome-Amat, Jaime, Barber, Domingo F., Ministerio de Economía y Competitividad (España), Agencia Estatal de Investigación (España), Instituto de Salud Carlos III, European Commission, Comunidad de Madrid, Díaz-Perales, Araceli [0000-0002-1093-3627], Escribese Maria M. [0000-0001-5057-5150], Garrido-Arandía, María [0000-0001-6114-5754], Obeso, David [0000-0001-7875-7327], Izquierdo-Alvarez, Elena [0000-0002-3355-2798], Tome-Amat, Jaime [0000-0003-4442-3649], Barber, Domingo F. [0000-0001-8824-5405], Díaz-Perales, Araceli, Escribese Maria M., Garrido-Arandía, María, Obeso, David, Izquierdo-Alvarez, Elena, Tome-Amat, Jaime, and Barber, Domingo F.

Abstract

Allergy is defined as a complex chronic inflammatory condition in which genetic and environmental factors are implicated. Sphingolipids are involved in multiple biological functions, from cell membrane components to critical signaling molecules. To date, sphingolipids have been studied in different human pathologies such as neurological disorders, cancer, autoimmunity, and infections. Sphingolipid metabolites, in particular, ceramide and sphingosine-1-phosphate (S1P), regulate a diverse range of cellular processes that are important in immunity and inflammation. Moreover, variations in the sphingolipid concentrations have been strongly associated with allergic diseases. This review will focus on the role of sphingolipids in the development of allergic sensitization and allergic inflammation through the activation of immune cells resident in tissues, as well as their role in barrier remodeling and anaphylaxis. The knowledge gained in this emerging field will help to develop new therapeutic options for allergic disorders.

Published
2021

15. Alternaria as an Inducer of Allergic Sensitization

Author

Ministerio de Economía y Competitividad (España), Comunidad de Madrid, Instituto de Salud Carlos III, European Commission, Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Barber, Domingo F. [0000-0001-8824-5405], Tome-Amat, Jaime [0000-0003-4442-3649], Garrido-Arandía, María [0000-0001-6114-5754], Díaz-Perales, Araceli [0000-0002-1093-3627], Hernandez-Ramirez, Guadalupe, Barber, Domingo F., Tome-Amat, Jaime, Garrido-Arandía, María, Díaz-Perales, Araceli, Ministerio de Economía y Competitividad (España), Comunidad de Madrid, Instituto de Salud Carlos III, European Commission, Hernandez-Ramirez, Guadalupe [0000-0003-3031-6754], Barber, Domingo F. [0000-0001-8824-5405], Tome-Amat, Jaime [0000-0003-4442-3649], Garrido-Arandía, María [0000-0001-6114-5754], Díaz-Perales, Araceli [0000-0002-1093-3627], Hernandez-Ramirez, Guadalupe, Barber, Domingo F., Tome-Amat, Jaime, Garrido-Arandía, María, and Díaz-Perales, Araceli

Abstract

Alternaria alternata is a saprophytic mold whose spores are disseminated in warm dry air, the typical weather of the Mediterranean climate region (from 30° to 45°), with a peak during the late summer and early autumn. Alternaria spores are known to be biological contaminants and a potent source of aeroallergens. One consequence of human exposure to Alternaria is an increased risk of developing asthma, with Alt a 1 as its main elicitor and a marker of primary sensitization. Although the action mechanism needs further investigation, a key role of the epithelium in cytokine production, TLR-activated alveolar macrophages and innate lymphoid cells in the adaptive response was demonstrated. Furthermore, sensitization to A. alternata seems to be a trigger for the development of co-sensitization to other allergen sources and may act as an exacerbator of symptoms and an elicitor of food allergies. The prevalence of A. alternata allergy is increasing and has led to expanding research on the role of this fungal species in the induction of IgE-mediated respiratory diseases. Indeed, recent research has allowed new perspectives to be considered in the assessment of exposure and diagnosis of fungi-induced allergies, although more studies are needed for the standardization of immunotherapy formulations.

Published
2021

16. Arabidopsis Leucine Rich Repeat-Malectin Receptor Kinases in immunity triggered by cellulose and mixed-linked glucan oligosaccharides

Author

Martín-Dacal, Marina, primary, Fernández-Calvo, Patricia, additional, Jiménez-Sandoval, Pedro, additional, López, Gemma, additional, Garrido-Arandía, María, additional, Rebaque, Diego, additional, del Hierro, Irene, additional, Torres, Miguel Ángel, additional, Kumar, Varun, additional, Berlanga, Diego José, additional, Mélida, Hugo, additional, Pacios, Luis F., additional, Santiago, Julia, additional, and Molina, Antonio, additional

Published
2022
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17. Structural Bases for the Allergenicity of Fra a 1.02 in Strawberry Fruits

Author

Ministerio de Economía y Competitividad (España), Agencia Estatal de Investigación (España), European Commission, Instituto de Salud Carlos III, Orozco-Navarrete, Begoña [0000-0001-8918-4683], Kaczmarska, Zuzanna [0000-0002-7668-1442], Dupeux, Florine [0000-0001-9532-5374], Garrido-Arandía, María [0000-0001-6114-5754], Pott, Delphine [0000-0002-2545-8777], Díaz-Perales, Araceli [0000-0002-1093-3627], Valpuesta, Victoriano [0000-0003-0922-6615], Merchante, Catharina [0000-0003-2214-9562], Orozco-Navarrete, Begoña, Kaczmarska, Zuzanna, Dupeux, Florine, Garrido-Arandía, María, Pott, Delphine, Díaz-Perales, Araceli, Casañal, Ana, Márquez, José A., Valpuesta, Victoriano, Merchante, Catharina, Ministerio de Economía y Competitividad (España), Agencia Estatal de Investigación (España), European Commission, Instituto de Salud Carlos III, Orozco-Navarrete, Begoña [0000-0001-8918-4683], Kaczmarska, Zuzanna [0000-0002-7668-1442], Dupeux, Florine [0000-0001-9532-5374], Garrido-Arandía, María [0000-0001-6114-5754], Pott, Delphine [0000-0002-2545-8777], Díaz-Perales, Araceli [0000-0002-1093-3627], Valpuesta, Victoriano [0000-0003-0922-6615], Merchante, Catharina [0000-0003-2214-9562], Orozco-Navarrete, Begoña, Kaczmarska, Zuzanna, Dupeux, Florine, Garrido-Arandía, María, Pott, Delphine, Díaz-Perales, Araceli, Casañal, Ana, Márquez, José A., Valpuesta, Victoriano, and Merchante, Catharina

Abstract

Although strawberries are highly appreciated fruits, their intake can induce allergic reactions in atopic patients. These reactions can be due to the patient's previous sensitization to the major birch pollen allergen Bet v 1, by which IgE generated in response to Bet v 1 cross-reacts with the structurally related strawberry Fra a 1 protein family. Fra a 1.02 is the most expressed paralog in ripe strawberries and is highly allergenic. To better understand the molecular mechanisms regulating this allergic response, we have determined the three-dimensional structure of Fra a 1.02 and four site-directed mutants that were designed based on their positions in potential epitopes. Fra a 1.02 and mutants conform to the START fold. We show that the cross-reactivity of all the mutant variants to IgE from patients allergic to Bet v 1 was significantly reduced without altering the conserved structural fold, so that they could potentially be used as hypoallergenic Fra a 1 variants for the generation of vaccines against strawberry allergy in atopic patients.

Published
2020

18. Dynamic plasticity of the lipid antigen-binding site of CD1d is crucially favoured by acidic pH and helper proteins

Author

Ministerio de Ciencia e Innovación (España), Comunidad de Madrid, Universidad Politécnica de Madrid, Banco Santander, European Commission, Agencia Estatal de Investigación (España), Cuevas-Zuviría, Bruno [0000-0003-1479-9442], Mínguez-Toral, Marina [0000-0001-7818-7360], Díaz-Perales, Araceli [0000-0002-1093-3627], Garrido-Arandía, María [0000-0001-6114-5754], Pacios, Luis F. [0000-0002-0585-4289], Cuevas-Zuviría, Bruno, Mínguez-Toral, Marina, Díaz-Perales, Araceli, Garrido-Arandía, María, Pacios, Luis F., Ministerio de Ciencia e Innovación (España), Comunidad de Madrid, Universidad Politécnica de Madrid, Banco Santander, European Commission, Agencia Estatal de Investigación (España), Cuevas-Zuviría, Bruno [0000-0003-1479-9442], Mínguez-Toral, Marina [0000-0001-7818-7360], Díaz-Perales, Araceli [0000-0002-1093-3627], Garrido-Arandía, María [0000-0001-6114-5754], Pacios, Luis F. [0000-0002-0585-4289], Cuevas-Zuviría, Bruno, Mínguez-Toral, Marina, Díaz-Perales, Araceli, Garrido-Arandía, María, and Pacios, Luis F.

Abstract

CD1 molecules present lipid antigens for recognition by T-cell receptors (TCRs). Although a reasonably detailed picture of the CD1-lipid-TCR interaction exists, the initial steps regarding lipid loading onto and exchange between CD1 proteins remain elusive. The hydrophobic nature of lipids and the fact that CD1 molecules are unable to extract lipids from membranes raise the need for the assistance of helper proteins in lipid trafficking. However, the experimental study of this traffic in the endosomal compartments at which it occurs is so challenging that computational studies can help provide mechanistic insight into the associated processes. Here we present a multifaceted computational approach to obtain dynamic structural data on the human CD1d isotype. Conformational dynamics analysis shows an intrinsic flexibility associated with the protein architecture. Electrostatic properties together with molecular dynamics results for CD1d complexes with several lipids and helper proteins unravel the high dynamic plasticity of the antigen-binding site that is crucially favoured by acidic pH and the presence of helper proteins.

Published
2020

19. A computational structural study on the DNA-protecting role of the tardigrade-unique Dsup protein

Author

Universidad Politécnica de Madrid, Banco Santander, European Commission, Centro de Supercomputación y Visualización de Madrid, Agencia Estatal de Investigación (España), Mínguez-Toral, Marina [0000-0001-7818-7360], Cuevas-Zuviría, Bruno [0000-0003-1479-9442], Garrido-Arandía, María [0000-0001-6114-5754], Pacios, Luis F. [0000-0002-0585-4289], Mínguez-Toral, Marina, Cuevas-Zuviría, Bruno, Garrido-Arandía, María, Pacios, Luis F., Universidad Politécnica de Madrid, Banco Santander, European Commission, Centro de Supercomputación y Visualización de Madrid, Agencia Estatal de Investigación (España), Mínguez-Toral, Marina [0000-0001-7818-7360], Cuevas-Zuviría, Bruno [0000-0003-1479-9442], Garrido-Arandía, María [0000-0001-6114-5754], Pacios, Luis F. [0000-0002-0585-4289], Mínguez-Toral, Marina, Cuevas-Zuviría, Bruno, Garrido-Arandía, María, and Pacios, Luis F.

Abstract

The remarkable ability of tardigrades to withstand a wide range of physical and chemical extremes has attracted a considerable interest in these small invertebrates, with a particular focus on the protective roles of proteins expressed during such conditions. The discovery that a tardigrade-unique protein named Dsup (damage suppressor) protects DNA from damage produced by radiation and radicals, has raised expectations concerning its potential applications in biotechnology and medicine. We present in this paper what might be dubbed a "computational experiment" on the Dsup-DNA system. By means of molecular modelling, calculations of electrostatic potentials and electric fields, and all-atom molecular dynamics simulations, we obtained a dynamic picture of the Dsup-DNA interaction. Our results suggest that the protein is intrinsically disordered, which enables Dsup to adjust its structure to fit DNA shape. Strong electrostatic attractions and high protein flexibility drive the formation of a molecular aggregate in which Dsup shields DNA. While the precise mechanism of DNA protection conferred by Dsup remains to be elucidated, our study provides some molecular clues of their association that could be of interest for further investigation in this line.

Published
2020

20. Arabidopsis immune responses triggered by cellulose‐ and mixed‐linked glucan‐derived oligosaccharides require a group of leucine‐rich repeat malectin receptor kinases.

Author

Martín‐Dacal, Marina, Fernández‐Calvo, Patricia, Jiménez‐Sandoval, Pedro, López, Gemma, Garrido‐Arandía, María, Rebaque, Diego, del Hierro, Irene, Berlanga, Diego José, Torres, Miguel Ángel, Kumar, Varun, Mélida, Hugo, Pacios, Luis F., Santiago, Julia, and Molina, Antonio

Subjects
LEUCINE, PATTERN perception receptors, IMMUNE response, OLIGOSACCHARIDES, ISOTHERMAL titration calorimetry, KINASES, PLANT cell walls, CELLULOSE synthase
Abstract

SUMMARY: The plant immune system perceives a diversity of carbohydrate ligands from plant and microbial cell walls through the extracellular ectodomains (ECDs) of pattern recognition receptors (PRRs), which activate pattern‐triggered immunity (PTI). Among these ligands are oligosaccharides derived from mixed‐linked β‐1,3/β‐1,4‐glucans (MLGs; e.g. β‐1,4‐D‐(Glc)2‐β‐1,3‐D‐Glc, MLG43) and cellulose (e.g. β‐1,4‐D‐(Glc)3, CEL3). The mechanisms behind carbohydrate perception in plants are poorly characterized except for fungal chitin oligosaccharides (e.g. β‐1,4‐d‐(GlcNAc)6, CHI6), which involve several receptor kinase proteins (RKs) with LysM‐ECDs. Here, we describe the isolation and characterization of Arabidopsis thaliana mutants impaired in glycan perception (igp) that are defective in PTI activation mediated by MLG43 and CEL3, but not by CHI6. igp1–igp4 are altered in three RKs – AT1G56145 (IGP1), AT1G56130 (IGP2/IGP3) and AT1G56140 (IGP4) – with leucine‐rich‐repeat (LRR) and malectin (MAL) domains in their ECDs. igp1 harbors point mutation E906K and igp2 and igp3 harbor point mutation G773E in their kinase domains, whereas igp4 is a T‐DNA insertional loss‐of‐function mutant. Notably, isothermal titration calorimetry (ITC) assays with purified ECD‐RKs of IGP1 and IGP3 showed that IGP1 binds with high affinity to CEL3 (with dissociation constant KD = 1.19 ± 0.03 μm) and cellopentaose (KD = 1.40 ± 0.01 μM), but not to MLG43, supporting its function as a plant PRR for cellulose‐derived oligosaccharides. Our data suggest that these LRR‐MAL RKs are components of a recognition mechanism for both cellulose‐ and MLG‐derived oligosaccharide perception and downstream PTI activation in Arabidopsis. Significance Statement: The mechanisms of perception of newly described cell‐wall derived oligosaccharides that trigger plant immunity are poorly characterized. Here we described three Arabidopsis thaliana receptor kinases (RK) with Leucine Rich Repeat (LRR) and Malectin (MAL) domains in their ectodomains (ECDs), which are components of a novel mechanism of both cellulose and mixed‐linked β‐glucan (MLGs)‐derived oligosaccharides perception and downstream immune activation. One of these RKs binds cellulose oligosaccharides indicating that is a plant pattern recognition receptor (PRR). [ABSTRACT FROM AUTHOR]

Published
2023
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21. Characterisation of a flavonoid ligand of the fungal protein Alt a 1

Author

Garrido-Arandía, María [0000-0001-6114-5754], Barber, Domingo F. [0000-0001-8824-5405], Díaz-Perales, Araceli [0000-0002-1093-3627], Pacios, Luis F. [0000-0002-0585-4289], Garrido-Arandía, María, Silva-Navas, J., Ramírez, Carmen, Cubells-Baeza, N., Gómez-Casado, Cristina, Barber, Domingo F., del Pozo, J. C., Gonzalez-Melendi, P., Pacios, Luis F., Díaz-Perales, Araceli, Garrido-Arandía, María [0000-0001-6114-5754], Barber, Domingo F. [0000-0001-8824-5405], Díaz-Perales, Araceli [0000-0002-1093-3627], Pacios, Luis F. [0000-0002-0585-4289], Garrido-Arandía, María, Silva-Navas, J., Ramírez, Carmen, Cubells-Baeza, N., Gómez-Casado, Cristina, Barber, Domingo F., del Pozo, J. C., Gonzalez-Melendi, P., Pacios, Luis F., and Díaz-Perales, Araceli

Abstract

Spores of pathogenic fungi are virtually ubiquitous and cause human disease and severe losses in crops. The endophytic fungi Alternaria species produce host-selective phytotoxins. Alt a 1 is a strongly allergenic protein found in A. alternata that causes severe asthma. Despite the well-established pathogenicity of Alt a 1, the molecular mechanisms underlying its action and physiological function remain largely unknown. To gain insight into the role played by this protein in the pathogenicity of the fungus, we studied production of Alt a 1 and its activity in spores. We found that Alt a 1 accumulates inside spores and that its release with a ligand is pH-dependent, with optimum production in the 5.0-6.5 interval. The Alt a 1 ligand was identified as a methylated flavonoid that inhibits plant root growth and detoxifies reactive oxygen species. We also found that Alt a 1 changes its oligomerization state depending on the pH of the surrounding medium and that these changes facilitate the release of the ligand. Based on these results, we propose that release of Alt a 1 should be a pathogenic target in approaches used to block plant defenses and consequently to favor fungal entry into the plant. © 2016 The Author(s).

Published
2016

23. Patterns of sensitization to inhalant allergens, Ole e 1 and Ole e 7 in children and adolescents born in the same area with different origin.

Author

Alvarez, Maria Somoza, Puche, Laura Victorio, Blanca-Lopez, Natalia, Ruano Pérez, Francisco Javier, Vázquez de La Torre Gaspar, Maria, Rojas, Isabel Torres, Lopez Sanchez, Jose Damian, Garrido-Arandia, Maria, Martín-Pedraza, Laura, Perales, Araceli Diaz, Villalba, M Teresa, Díez, Gabriela Canto, and Blanca, Miguel

Published
2020
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26. Arabidopsis immune responses triggered by cellulose‐ and mixed‐linked glucan‐derived oligosaccharides require a group ofleucine‐rich repeat malectinreceptor kinases

Author

Fisiologia Vegetal, Martín Dacal, Marina, Fernández Calvo, Patricia, Jiménez Sandoval, Pedro, López, Gemma, Garrido Arandía, María, Rebaque, Diego, Hierro, Irene del, Berlanga, Diego José, Torres, Miguel Ángel, Kumar, Varun, Mélida Martínez, Hugo, Pacios, Luis F., Santiago, Julia, Molina, Antonio, Fisiologia Vegetal, Martín Dacal, Marina, Fernández Calvo, Patricia, Jiménez Sandoval, Pedro, López, Gemma, Garrido Arandía, María, Rebaque, Diego, Hierro, Irene del, Berlanga, Diego José, Torres, Miguel Ángel, Kumar, Varun, Mélida Martínez, Hugo, Pacios, Luis F., Santiago, Julia, and Molina, Antonio

Abstract

[EN] The plant immune system perceives a diversity of carbohydrate ligands from plant and microbial cell walls through the extracellular ectodomains (ECDs) of pattern recognition receptors (PRRs), which activate pattern-triggered immunity (PTI). Among these ligands are oligosaccharides derived from mixed-linked b- 1,3/b-1,4-glucans (MLGs; e.g. b-1,4-D-(Glc)2-b-1,3-D-Glc, MLG43) and cellulose (e.g. b-1,4-D-(Glc)3, CEL3). The mechanisms behind carbohydrate perception in plants are poorly characterized except for fungal chitin oligosaccharides (e.g. b-1,4-D-(GlcNAc)6, CHI6), which involve several receptor kinase proteins (RKs) with LysM-ECDs. Here, we describe the isolation and characterization of Arabidopsis thaliana mutants impaired in glycan perception (igp) that are defective in PTI activation mediated by MLG43 and CEL3, but not by CHI6. igp1–igp4 are altered in three RKs – AT1G56145 (IGP1), AT1G56130 (IGP2/IGP3) and AT1G56140 (IGP4) – with leucine-rich-repeat (LRR) and malectin (MAL) domains in their ECDs. igp1 harbors point mutation E906K and igp2 and igp3 harbor point mutation G773E in their kinase domains, whereas igp4 is a T-DNA insertional loss-of-function mutant. Notably, isothermal titration calorimetry (ITC) assays with purified ECDRKs of IGP1 and IGP3 showed that IGP1 binds with high affinity to CEL3 (with dissociation constant KD = 1.19 0.03 lM) and cellopentaose (KD = 1.40 0.01 lM), but not to MLG43, supporting its function as a plant PRR for cellulose-derived oligosaccharides. Our data suggest that these LRR-MAL RKs are components of a recognition mechanism for both cellulose- and MLG-derived oligosaccharide perception and downstream PTI activation in Arabidopsis.

27. Arabidopsis immune responses triggered by cellulose- and mixed-linked glucan-derived oligosaccharides require a group of leucine-rich repeat malectin receptor kinases

Author

Marina Martín‐Dacal, Patricia Fernández‐Calvo, Pedro Jiménez‐Sandoval, Gemma López, María Garrido‐Arandía, Diego Rebaque, Irene del Hierro, Diego José Berlanga, Miguel Ángel Torres, Varun Kumar, Hugo Mélida, Luis F. Pacios, Julia Santiago, Antonio Molina, Agencia Estatal de Investigación (España), Ministerio de Ciencia, Innovación y Universidades (España), Comunidad de Madrid, Swiss National Science Foundation, Martín-Dacal, Marina, Fernández-Calvo, Patricia, Jiménez-Sandoval, Pedro, Garrido-Arandía, María, Rebaque, Diego, Del Hierro, Irene, Mélida, Hugo, Pacios, Luis F., Santiago, Julia, and Molina, Antonio

Subjects
Mixed-linked glucans (MLGs), Pattern recognition receptors (PRRs), Arabidopsis thaliana, Arabidopsis/metabolism, Protein Serine-Threonine Kinases/metabolism, Arabidopsis Proteins/genetics, Arabidopsis Proteins/metabolism, Leucine/metabolism, Glucans/metabolism, Cellulose/metabolism, Plant Immunity/genetics, Plants/metabolism, Oligosaccharides/metabolism, cellulose, immunity, leucine-reach repeat/Malectin receptor kinase (LRR-MAL RK), mixed-linked glucans (MLGs), oligosaccharides, pattern recognition receptors (PRRs), Genetics, Immunity, Oligosaccharides, Leucine-reach repeat/Malectin receptor kinase (LRR-MAL RK), Cell Biology, Plant Science, Cellulose
Abstract

18 Päg., The plant immune system perceives a diversity of carbohydrate ligands from plant and microbial cell walls through the extracellular ectodomains (ECDs) of pattern recognition receptors (PRRs), which activate pattern-triggered immunity (PTI). Among these ligands are oligosaccharides derived from mixed-linked β-1,3/β-1,4-glucans (MLGs; e.g. β-1,4-D-(Glc)2 -β-1,3-D-Glc, MLG43) and cellulose (e.g. β-1,4-D-(Glc)3 , CEL3). The mechanisms behind carbohydrate perception in plants are poorly characterized except for fungal chitin oligosaccharides (e.g. β-1,4-d-(GlcNAc)6 , CHI6), which involve several receptor kinase proteins (RKs) with LysM-ECDs. Here, we describe the isolation and characterization of Arabidopsis thaliana mutants impaired in glycan perception (igp) that are defective in PTI activation mediated by MLG43 and CEL3, but not by CHI6. igp1-igp4 are altered in three RKs - AT1G56145 (IGP1), AT1G56130 (IGP2/IGP3) and AT1G56140 (IGP4) - with leucine-rich-repeat (LRR) and malectin (MAL) domains in their ECDs. igp1 harbors point mutation E906K and igp2 and igp3 harbor point mutation G773E in their kinase domains, whereas igp4 is a T-DNA insertional loss-of-function mutant. Notably, isothermal titration calorimetry (ITC) assays with purified ECD-RKs of IGP1 and IGP3 showed that IGP1 binds with high affinity to CEL3 (with dissociation constant KD = 1.19 ± 0.03 μm) and cellopentaose (KD = 1.40 ± 0.01 μM), but not to MLG43, supporting its function as a plant PRR for cellulose-derived oligosaccharides. Our data suggest that these LRR-MAL RKs are components of a recognition mechanism for both cellulose- and MLG-derived oligosaccharide perception and downstream PTI activation in Arabidopsis., This work was supported by grant RTI2018-096975-B-I00 from the Spanish Ministry of Science, Innovation and Universities to AM and grant PID-2021-126006OB-100 from the Spanish Ministry of Science and Innovation to AM. This work has also been financially supported by the ‘Severo Ochoa (SO) Programme for Centres of Excellence in R&D’ from the Agencia Estatal de Investigación (AEI) of Spain (grants SEV-2016-0672 (2017-2021) and CEX2020-000999-S (2022-2025) to the CBGP). In the frame of the SO program, HM and PF-C were supported with postdoctoral fellowships. MM-D, DJB and DR were recipients of PhD Fellows PRE2019-088120 and PRE2019-091276 (SEV-2016-0672) from AEI, and IND2017/BIO-7800 from Madrid Regional Government, respectively. Research in the lab of JS was financially supported by the University of Lausanne, the European Research Council (ERC) (grant agreement no. 716358) and the Swiss National Science Foundation (grant no. 310030_204526)., With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2020‐000999‐S)

Published
2023

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