Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients
Wheat-dependent exercise-induced anaphylaxis (WDEIA) is one of the most severe forms of wheat allergy. It occurs in patients when they exercise after ingesting wheat-containing foods. Nowadays, the only possible alternative for WDEIA patients is to avoid such foods. This study investigated the potential of six RNA of interference (RNAi) wheat lines with low-prolamin content as alternatives for WDEIA patients. For that purpose, a high performance-liquid chromatography (HPLC) analysis was performed to evaluate differences in gluten protein fractions among these lines. Next, western blots were conducted to measure the immunoglobulin E (IgE) reactivity to wheat proteins in sera from five WDEIA patients. Additionally, monoclonal antibodies (moAb) recognition sites and the IgE binding sites were searched in all peptides identified by LC-MS/MS after protein digestion. The results showed a 61.4%–81.2% reduction in the gliadin content of the RNAi lines, accompanied by an increase in their high-molecular weight (HMW) glutenin content compared to the wild type bread wheat line (WT). In all cases, the reduction in gliadin content correlated with a decrease in IgE reactivity observed in the sera of WDEIA patients, highlighting the E82 and H320 lines. These two RNAi lines exhibited a ≤90% reduction in IgE reactivity. This reduction could be attributed to an absence of IgE binding sites associated with α- and ω5-gliadins, which were present in the WT. Overall, these lines offer a potential alternative for foodstuff for individuals with WDEIA.
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Format: | artículo biblioteca |
Language: | English |
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Frontiers Media
2023
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Subjects: | RNAi, Gluten, IgE, Wheat, Wheat-dependent exercise-induced anaphylaxis (WDEIA), Reverse-phase high-performance liquid chromatography (RP-HPLC), Non-gluten proteins (NGPs), |
Online Access: | http://hdl.handle.net/10261/353228 |
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dig-ias-es-10261-3532282024-04-09T07:15:49Z Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients Guzmán-López, María H. Ruipérez, Violeta Marín-Sanz, Miriam Ojeda-Fernández, Isabel Ojeda-Fernández, Pedro Garrote, José Antonio Arranz, Eduardo Barro Losada, Francisco Ministerio de Ciencia, Innovación y Universidades (España) Agencia Estatal de Investigación (España) Ministerio de Ciencia e Innovación (España) European Commission Junta de Andalucía Consejo Superior de Investigaciones Científicas (España) RNAi Gluten IgE Wheat Wheat-dependent exercise-induced anaphylaxis (WDEIA) Reverse-phase high-performance liquid chromatography (RP-HPLC) Non-gluten proteins (NGPs) Wheat-dependent exercise-induced anaphylaxis (WDEIA) is one of the most severe forms of wheat allergy. It occurs in patients when they exercise after ingesting wheat-containing foods. Nowadays, the only possible alternative for WDEIA patients is to avoid such foods. This study investigated the potential of six RNA of interference (RNAi) wheat lines with low-prolamin content as alternatives for WDEIA patients. For that purpose, a high performance-liquid chromatography (HPLC) analysis was performed to evaluate differences in gluten protein fractions among these lines. Next, western blots were conducted to measure the immunoglobulin E (IgE) reactivity to wheat proteins in sera from five WDEIA patients. Additionally, monoclonal antibodies (moAb) recognition sites and the IgE binding sites were searched in all peptides identified by LC-MS/MS after protein digestion. The results showed a 61.4%–81.2% reduction in the gliadin content of the RNAi lines, accompanied by an increase in their high-molecular weight (HMW) glutenin content compared to the wild type bread wheat line (WT). In all cases, the reduction in gliadin content correlated with a decrease in IgE reactivity observed in the sera of WDEIA patients, highlighting the E82 and H320 lines. These two RNAi lines exhibited a ≤90% reduction in IgE reactivity. This reduction could be attributed to an absence of IgE binding sites associated with α- and ω5-gliadins, which were present in the WT. Overall, these lines offer a potential alternative for foodstuff for individuals with WDEIA. The project was funded by the Spanish Agencia Estatal de Investigación (PID2019-110847RB-I00, TED2021-129733B-I00 and PID2022-142139OB-I00, MCIN/AEI/10.13039/501100011033), European Union (“NextGenerationEU”/PRTR), Junta de Andalućıa (QUAL21_023 IAS) and the WheatNet (“Conexión TRIGO”) of the Spanish National Research Council (CSIC). Peer reviewed 2024-04-09T07:15:49Z 2024-04-09T07:15:49Z 2023 artículo Frontiers in Nutrition 10: 1319888 (2023) http://hdl.handle.net/10261/353228 10.3389/fnut.2023.1319888 2296-861X en #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-110847RB-I00/ES/REDISEÑO DE LAS PROTEINAS INMUNOGENICAS DE TRIGO RELACIONADAS CON LA CELIAQUIA MEDIANTE CRISPR%2FCAS/ info:eu-repo/grantAgreement/AEI//TED2021-129733B-I00 info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2022-142139OB-I00/ES/ESTRATEGIAS CRISPR%2FCAS PARA SUSTITUIR LOS PEPTIDOS INMUNOGENICOS PRESENTES EN LOS GENES DE GLIADINA DE TRIGO POR VERSIONES NO INMUNOGENICAS PRESERVANDO LA CALIDAD/ Publisher's version https://doi.org/10.3389/fnut.2023.1319888 Sí open application/pdf Frontiers Media |
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RNAi Gluten IgE Wheat Wheat-dependent exercise-induced anaphylaxis (WDEIA) Reverse-phase high-performance liquid chromatography (RP-HPLC) Non-gluten proteins (NGPs) RNAi Gluten IgE Wheat Wheat-dependent exercise-induced anaphylaxis (WDEIA) Reverse-phase high-performance liquid chromatography (RP-HPLC) Non-gluten proteins (NGPs) |
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RNAi Gluten IgE Wheat Wheat-dependent exercise-induced anaphylaxis (WDEIA) Reverse-phase high-performance liquid chromatography (RP-HPLC) Non-gluten proteins (NGPs) RNAi Gluten IgE Wheat Wheat-dependent exercise-induced anaphylaxis (WDEIA) Reverse-phase high-performance liquid chromatography (RP-HPLC) Non-gluten proteins (NGPs) Guzmán-López, María H. Ruipérez, Violeta Marín-Sanz, Miriam Ojeda-Fernández, Isabel Ojeda-Fernández, Pedro Garrote, José Antonio Arranz, Eduardo Barro Losada, Francisco Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients |
description |
Wheat-dependent exercise-induced anaphylaxis (WDEIA) is one of the most severe forms of wheat allergy. It occurs in patients when they exercise after ingesting wheat-containing foods. Nowadays, the only possible alternative for WDEIA patients is to avoid such foods. This study investigated the potential of six RNA of interference (RNAi) wheat lines with low-prolamin content as alternatives for WDEIA patients. For that purpose, a high performance-liquid chromatography (HPLC) analysis was performed to evaluate differences in gluten protein fractions among these lines. Next, western blots were conducted to measure the immunoglobulin E (IgE) reactivity to wheat proteins in sera from five WDEIA patients. Additionally, monoclonal antibodies (moAb) recognition sites and the IgE binding sites were searched in all peptides identified by LC-MS/MS after protein digestion. The results showed a 61.4%–81.2% reduction in the gliadin content of the RNAi lines, accompanied by an increase in their high-molecular weight (HMW) glutenin content compared to the wild type bread wheat line (WT). In all cases, the reduction in gliadin content correlated with a decrease in IgE reactivity observed in the sera of WDEIA patients, highlighting the E82 and H320 lines. These two RNAi lines exhibited a ≤90% reduction in IgE reactivity. This reduction could be attributed to an absence of IgE binding sites associated with α- and ω5-gliadins, which were present in the WT. Overall, these lines offer a potential alternative for foodstuff for individuals with WDEIA. |
author2 |
Ministerio de Ciencia, Innovación y Universidades (España) |
author_facet |
Ministerio de Ciencia, Innovación y Universidades (España) Guzmán-López, María H. Ruipérez, Violeta Marín-Sanz, Miriam Ojeda-Fernández, Isabel Ojeda-Fernández, Pedro Garrote, José Antonio Arranz, Eduardo Barro Losada, Francisco |
format |
artículo |
topic_facet |
RNAi Gluten IgE Wheat Wheat-dependent exercise-induced anaphylaxis (WDEIA) Reverse-phase high-performance liquid chromatography (RP-HPLC) Non-gluten proteins (NGPs) |
author |
Guzmán-López, María H. Ruipérez, Violeta Marín-Sanz, Miriam Ojeda-Fernández, Isabel Ojeda-Fernández, Pedro Garrote, José Antonio Arranz, Eduardo Barro Losada, Francisco |
author_sort |
Guzmán-López, María H. |
title |
Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients |
title_short |
Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients |
title_full |
Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients |
title_fullStr |
Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients |
title_full_unstemmed |
Identification of RNAi hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients |
title_sort |
identification of rnai hypoallergic bread wheat lines for wheat-dependent exercise-induced anaphylaxis patients |
publisher |
Frontiers Media |
publishDate |
2023 |
url |
http://hdl.handle.net/10261/353228 |
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