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)

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.Peer reviewe

Towards a deeper understanding of the pathophysiology of allergic diseases

La prevalencia de enfermedades alérgicas ha crecido notablemente a lo largo de las últimas décadas, afectando negativamente a la calidad de vida de los pacientes, así como incrementando los costes médicos de los sistemas públicos de salud. La inmunoterapia hiposensibilizante (AIT) es el tratamiento más prometedor de cara a poder erradicar completamente estas enfermedades, pero su uso todavía no está extendido debido a los bajos ratios beneficio/riesgo que presentan las formulaciones actuales. Por tanto, desarrollar nuevas AIT y mejorar sus protocolos de administración resulta vital. Debido a esto, el primer objetivo de esta tesis ha consistido en estudiar los mecanismos de sensibilización a alérgenos respiratorios, tomando Alt a 1 (el alérgeno mayor de Alternaria alternata) como modelo. Mediante el uso de ratones de experimentación, se ha demostrado que Alt a 1 no necesita adyuvantes para promover fenotipos alérgicos in vivo. En su lugar, es capaz de unirse a receptores TLR4 presentes en células con un fenotipo similar al de los macrófagos alveolares. Esto podría estar relacionado con su capacidad para activar la ruta proinflamatoria NF-κB y la señalización a través del inflamasoma (↑ caspasa-1 bioactiva), lo que promovería la infiltración a nivel pulmonar de diferentes células del sistema inmune (Siglec-F+, F4/80+ y CD3+, respectivamente). En conjunto, todo ello permite la sensibilización del individuo y la producción de anticuerpos frente al alérgeno. El segundo objetivo de esta tesis ha sido profundizar en nuestro conocimiento del proceso de sensibilización alérgica a los alimentos, tomando como modelo la sensibilización a Pru p 3 (alérgeno mayor del melocotón) a través de la piel. Con ello se ha demostrado que la exposición constante a cremas depilatorias provoca a largo plazo un primado de la ruta NLRP3 in vivo, y que esta ruta es fundamental para promover la sensibilización a Pru p 3. A mayores, la exposición al alérgeno (pero no la depilación) promueven el remodelado de la piel, así como la infiltración de células CD45+ no solo en el área sensibilizada (la piel), sino a lo largo de diferentes tejidos (pulmones e intestinos), sistematizando de este modo la respuesta. Finalmente, se ha estudiado la habilidad de partículas similares a virus (VLPs) derivadas del virus del mosaico del nabo (TuMV) como plataformas para ser usadas en el tratamiento de las alergias, usando la alergia a Pru p 3 como modelo. La caracterización inicial de la fórmula (VLP-Pru p 3 con su ligando lípido, esto es, VLP-Complex) confirmó su capacidad para inducir la proliferación de células inmunológicas humanas in vitro. Además, la fórmula fue transportada por células epiteliales intestinales, sin afectar negativamente a la integridad de la barrera. Finalmente, la administración sublingual de VLP-Complex fue capaz de reducir algunos marcadores serológicos asociados con respuestas alérgicas en ratones sensibilizados a Pru p 3, mejorando también la infiltración de células CD45+ en la piel de los mismos especímenes. Estos resultados no se acompañaron de efectos secundarios detectables a nivel macroscópico, nefrítico o hepático. En resumen, los resultados presentados en esta tesis permiten profundizar en el conocimiento de los mecanismos moleculares de la sensibilización a Alt a 1 y Pru p 3. Asimismo, presentan una nueva formulación (VLP-Complex) candidata a ser estudiada en el futuro como AIT para el tratamiento de la alergia a Pru p 3. ABSTRACT The prevalence of allergic diseases has constantly risen over the last decades, significantly impacting the quality of life of the patients, and increasing medical costs of public healthcare economies. Allergen-specific immunotherapy (AIT) is the most promising treatment that can reverse the allergic phenotype of an individual, but its use is not extended because of insufficient benefit/risk ratios of current formulations. Developing new AIT strategies and improving administration protocols is vital to overcome the problems associated with allergic diseases. In light of the above, the first objective of this thesis was to study allergic sensitization to airborne allergens, using Alt a 1 (Alternaria alternata’s major allergen) as a model. Using a murine model of the pathology, we demonstrated that Alt a 1 does not need of an exogenous adjuvant to promote allergic sensitization in vivo. Instead, it is able to bind to TLR4 molecules in alveolar macrophage-like cells present in healthy lungs. This might be related with its ability to activate downstream NF-κB and inflammasome (↑ bioactive caspase-1) signaling, which subsequently promote the infiltration of the lung with cells from immune origin (Siglec-F+, F4/80+ and CD3+ cells, respectively), to promote sensitizing responses and antibody production. The second objective of this thesis has been to deepen our knowledge regarding allergic sensitization to food, using epicutaneous sensitization to Pru p 3 (peach’s major allergen) as a model. By studying the effects of long-term continued depilation on the skin, we have proven that it leads to priming of the NLRP3 pathway in vivo, and that this route is vital to promote sensitization to Pru p 3. 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 (lungs and gut), furtherly validating the systemization of the response. Finally, we studied the ability of virus-like particles (VLPs) derived from Turnip mosaic virus (TuMV) as presenting platforms to be used in the treatment of allergic pathologies, using allergy to Pru p 3 as a model. Initial in vitro characterization of the formula (VLP-Pru p 3 with its associated lipid ligand, i.e., VLP-Complex) confirmed that it was able to induce proliferation of human immune cells. In addition, VLP-Complex was also able to be transported by intestinal epithelial cells, without affecting the monolayer integrity. Finally, sublingual administration of VLP-Complex could effectively reduce some serological markers associated with allergic responses in Pru p 3-sensitized mice, as well as CD45+ infiltration in the skin. Noteworthy, no associated macroscopic, nephritic, or hepatic toxicity was detected, as assessed by weight, blood urea nitrogen (BUN) and galectin-3 analyses, respectively. To sum up, the results presented in this thesis have helped to increase our knowledge regarding how sensitization to Alt a 1 and Pru p 3 works. In addition, it presents a new putative AIT formulation (VLP-Complex), that can be studied and furtherly optimized to treat Pru p 3 allergy in a future

Plant non-specific lipid transfer proteins: An overview

13 Pág.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.KM was supported through funding by the Ministry of Higher Education and Scientific Research of Tunisia (CP_2019–2022), ZGK was funded by the Community of Madrid (FOODAL-CM_S2018/BAAA-4574). DPC was granted by Universidad Politécnica de Madrid and Banco Santander for a predoctoral Programa Propio grant. GHR was funded by European Commission (H2020-NMBP-X-KET-2017-768641 AllerScreening). JTA was funded by Instituto de Salud Carlos III (ISCIII) co-founded by FEDER Thematic Networks and Cooperative Research Centers: ARADYAL (RD16/0006/0003). This work was also funded by the Spanish Ministry of Science and Innovation (LISSENTRA; PID2020-113629RB-I00).Peer reviewe

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

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.This research was funded by the Spanish Government (MINECO, grant BIO2017-84548- R); Instituto de Salud Carlos III (ISCIII) co-founded by FEDER Thematic Networks and Cooperative Research Centers: ARADYAL (RD16/0006/0003; RD16/0006/0013) and the Community of Madrid (FOODAL-CM_S2018/BAAA-4574). DPC was funded by Universidad Politécnica de Madrid and Banco Santander with a predoctoral Programa Propio grant. ARS was funded from the Community of Madrid included in the project FOOD-AL (FOODAL-CM_S2018/BAAA-4574). GHR was funded by European Comission (H2020-NMBP-X-KET-2017-768641—AllerScreening). JTA was funded from Instituto de Salud Carlos III (ISCIII) co-founded by FEDER Thematic Networks and Cooperative Research Centers: ARADYAL (RD16/0006/0003).Peer reviewe

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

12 Pág. Centro de Biotecnología y Genómica de Plantas (CBGP)This research was funded by the Spanish Ministry of Science and Innovation through the project LISENTRA, granted by the Spanish Research State Agency (PID2020-113629RB00/AEI/10.13039/501100011033). GH-R was granted by funding from the European Commission through the project AllerScreening, granted within the R&D framework programme Horizon2020 (H2020-NMBP-X- KET-2017-768641). DP-C was granted by Universidad Politécnica de Madrid and Banco Santander for a predoctoral Programa Propio grant. JT-A and ZG-K were granted by funding from the Community of Madrid included in the project FOODAL (FOODAL-CM; S2018/BAA-4574) co-funded by ESF and ERDF R&D projects call Tecnologías 2018. This work was also supported by Instituto de Salud Carlos III (ISCIII) co-funded by FEDER Thematic Networks and Cooperative Research Centers: ARADYAL (RD16/0006/0003; RD16/0006/0013).Peer reviewe

The key to the allergenicity of lipid transfer protein (LTP) ligands: A structural characterization

Plant lipid transfer proteins are a large family that can be found in all land plants. They have a hydrophobic cavity that allows them to harbor lipids and facilitates their traffic between membranes. However, in humans, this plant protein family is responsible for the main food allergies in the Mediterranean area. Nevertheless, not only the protein itself but also its ligand is relevant for allergic sensitization. The main aim of the present work is to analyse the natural ligands carried by four allergenic LTPs (Tri a 14, Art v 3, Par j 2, and Ole e 7), compared with the previously identified ligand of Pru p 3 (CPT-PHS ligand), and clarify their role within the immunological reactions. Results showed that the ligands of the LTPs studied shared a chemical identity, in which the presence of a polar head was essential to the protein-ligand binding. This ligand was transported through a skin cellular model, and phosphorylated phytosphingosine could be detected as result of cell metabolism. Since sphingosine kinase 1 was overexpressed in keratinocytes incubated with the LTP-ligand complex, this enzyme might be responsible for the phosphorylation of the phytosphingosine fraction of the CPT-PHS ligand. This way, phytosphingosine-1-phosphate could be mimicking the role of the human inflammatory mediator sphingosine-1-phosphate, explaining why LTPs are associated with more severe allergic responses. In conclusion, this work contributes to the understanding of the chemical nature and behavior of lipid ligands carried by allergens, which would help to gain insight into their role during allergic sensitization

Memorias del primer Simposio Nacional de Ciencias Agronómicas

Primer simposio nacional de Ciencias Agronómicas: El renacer del espacio de discusión científica para el Agro colombiano

Espacios y destinos turísticos en tiempos de globalización y crisis

2 volúmenesXII Coloquio de Geografía del Turismo, Ocio y Recreación de la Asociación de Geógrafos Españoles. Colmenarejo (Madrid), del 17 al 19 de junio de 2010.Este libro ha sido editado con la colaboración económica del Ministerio de Ciencia e Innovación (ref. CS02010-10416-E)

Memorias del primer Simposio Nacional de Ciencias Agronómicas

Primer simposio nacional de Ciencias Agronómicas: El renacer del espacio de discusión científica para el Agro colombiano