1. Intrinsic alterations in peripheral neutrophils from cystic fibrosis newborn piglets
- Author
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Mustapha Berri, Isabelle Fleurot, Sandrine Melo, Laura Soler, Ignacio S. Caballero, Ana-Paula Teixeira-Gomes, Valérie Labas, Nicolas Cenac, Mustapha Si-Tahar, Julien Pujo, Andrea Bähr, Déborah Bréa, Claire Chevaleyre, Nikolai Klymiuk, Antoine Guillon, SEGUIN, Nathalie, Centre d’Etude des Pathologies Respiratoires (CEPR), UMR 1100 (CEPR), Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Biosynthèse & Toxicité des Mycotoxines (ToxAlim-BioToMyc), ToxAlim (ToxAlim), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Ecole d'Ingénieurs de Purpan (INPT - EI Purpan), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Infectiologie et Santé Publique (UMR ISP), Université de Tours (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire d'Analyses des Interactions Moléculaires, Plate-forme d'Analyses intégrative des Biomarqueurs cellulaires et moléculaires (PAIB), UMR6175, Institut National de la Recherche Agronomique (INRA), Institut de Recherche en Santé Digestive (IRSD ), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Gene Center and Center for Innovation Medical Models (CiMM), Ludwig-Maximilians University [Munich] (LMU), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours), Vaincre la Mucoviscidose [grant numbers RF20130500925 and RF2015050357, Region Centre (France) [grant number 320 0 0604 ]., Metatoul- Lipidomique Core Facility-MetaboHub, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Tours, Université de Tours-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Ludwig Maximilians University of Munich, Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), and Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
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0301 basic medicine ,Pulmonary and Respiratory Medicine ,Male ,Proteomics ,Swine ,Neutrophils ,[SDV.MHEP.PHY] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO] ,[SDV]Life Sciences [q-bio] ,Inflammation ,medicine.disease_cause ,Cystic fibrosis ,Flow cytometry ,03 medical and health sciences ,0302 clinical medicine ,Immunophenotyping ,medicine ,[SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO] ,Animals ,CFTR ,chemistry.chemical_classification ,Reactive oxygen species ,Proteomic Profile ,Lung ,medicine.diagnostic_test ,Pseudomonas aeruginosa ,business.industry ,Models, Theoretical ,medicine.disease ,Pig model ,Molecular biology ,3. Good health ,[SDV] Life Sciences [q-bio] ,Disease Models, Animal ,030104 developmental biology ,medicine.anatomical_structure ,030228 respiratory system ,chemistry ,Animals, Newborn ,ICM-MS ,Pediatrics, Perinatology and Child Health ,Fatty Acids, Unsaturated ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Female ,medicine.symptom ,business ,Reactive Oxygen Species - Abstract
International audience; Background: The hallmark of the cystic fibrosis (CF) lung disease is a neutrophil dominated lung environment that is associated to chronic lung tissue destruction and ultimately the patient's death. It is unclear whether the exacerbated neutrophil response is primary related to a defective CFTR or rather secondary to chronic bacterial colonization and inflammation. Here, we hypothesized that CF peripheral blood neutrophils present intrinsic alteration at birth before the start of an inflammatory process.Methods: Peripheral blood neutrophils were isolated from newborn CFTR+/+ and CFTR-/- piglets. Neutrophils immunophenotype was evaluated by flow cytometry. Lipidomic and proteomic profile were characterized by liquid chromatography/tandem mass spectrometry (LC-MS/MS), intact cell matrix-assisted laser desorption/ionization mass spectrometry (ICM-MS) followed by top-down high-resolution mass spectrometry (HRMS), respectively. The ability of CF neutrophils to kill pseudomonas aeruginosa was also evaluated.Results: Polyunsaturated fatty acid metabolites analysis did not show any difference between CFTR+/+ and CFTR-/- neutrophils. On the other hand, a predictive mathematical model based on the ICM-MS proteomic profile was able to discriminate between both genotypes. Top-down proteomic analysis identified 19 m/z differentially abundant masses that corresponded mainly to proteins related to the antimicrobial response and the generation of reactive oxygen species (ROS). However, no alteration in the ability of CFTR-/- neutrophils to kill pseudomonas aeruginosa in vitro was observed.Conclusions: ICM-MS demonstrated that CFTR-/- neutrophils present intrinsic alterations already at birth, before the presence of any infection or inflammation.
- Published
- 2020
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