Contributors: 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 de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT); Institut National Polytechnique (Toulouse) (Toulouse INP); Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP); Université de Toulouse (UT)-Ecole d'Ingénieurs de Purpan (INP - PURPAN); Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Toulouse III - Paul Sabatier (UT3); Université de Toulouse (UT)-Université de Toulouse (UT)-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 de Toulouse (UT)-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
نبذة مختصرة : 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.
Processing Request
No Comments.