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15 results on '"MESH : Epithelial Cells"'

1. Candida albicans is able to use M cells as a portal of entry across the intestinal barrier in vitro

Author

Albac, Sandrine, Schmitz, Antonin, Lopez-Alayon, Carolina, d'Enfert, Christophe, Sautour, Marc, Ducreux, Amandine, Labruère-Chazal, Catherine, Laue, Michael, Holland, Gudrun, Bonnin, Alain, Dalle, Frederic, Agroécologie [Dijon], Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Biologie et Pathogénicité fongiques, Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), Laboratoire de parasitologie mycologie (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Institut de Mathématiques de Bourgogne [Dijon] (IMB), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Bourgogne (UB), Robert Koch Institute [Berlin] (RKI), Agence Nationale de la Recherche. Grant Number: ANR-08-MIEN- 033-01 BIOASTER-Sanofi-Alliance pour les Sciences de la Vie et de la Santé (AVIESAN) Investissement d'Avenir. Grant Number: ANR-10-AIRT-03, ANR-08-MIEN-0033,KANJI,Colonisation et invasion de la muqueuse digestive par le champignon pathogène de l'homme Candida albicans(2008), ANR-10-AIRT-0005,NANOELEC,NANOELEC(2010), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris], Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS), ANR: ANR-10-AIRT-05,Programme Investissements d’Avenir, Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement, Institut National de la Recherche Agronomique ( INRA ) -Institut Pasteur [Paris], Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand ( CHU Dijon ), Institut de Mathématiques de Bourgogne [Dijon] ( IMB ), Université de Bourgogne ( UB ) -Centre National de la Recherche Scientifique ( CNRS ), Robert Koch Institute [Berlin] ( RKI ), ANR-08-MIEN-0033,KANJI,Colonisation et invasion de la muqueuse digestive par le champignon pathogène de l'homme Candida albicans ( 2008 ), ANR : ANR-10-AIRT-05,Programme Investissements d’Avenir, Biologie et Pathogénicité fongiques (BPF), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP), and Université de Bourgogne (UB)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH : Cell Line, MESH : Coculture Techniques, MESH : Microscopy, Fluorescence, [SDV]Life Sciences [q-bio], MESH: Microscopy, Fluorescence, MESH: Candidemia, Peyer's Patches, Mucosal immunity, Invasion, Candida albicans, MESH : Host-Pathogen Interactions, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, B-Lymphocytes, Endocytosis, MESH : Epithelial Cells, MESH : Peyer's Patches, MESH: Epithelial Cells, Host-Pathogen Interactions, MESH: Endocytosis, Adhesion, Transcytosis, Infection, MESH : Microscopy, Electron, Scanning, MESH : Candida albicans, MESH: Microscopy, Electron, Scanning, MESH : Endocytosis, MESH : B-Lymphocytes, Cell Line, MESH: Cell Adhesion, MESH: Coculture Techniques, MESH: B-Lymphocytes, Cell Adhesion, MESH : Candidemia, Humans, MESH: Humans, [ SDV ] Life Sciences [q-bio], MESH: Candida albicans, MESH : Humans, MESH: Host-Pathogen Interactions, Candidemia, Epithelial Cells, Coculture Techniques, MESH: Cell Line, Gastrointestinal Tract, MESH : Cell Adhesion, Microscopy, Fluorescence, Microscopy, Electron, Scanning, MESH: Peyer's Patches, MESH: Gastrointestinal Tract, MESH : Gastrointestinal Tract
Abstract

International audience; Candida albicans is the most frequent yeast responsible for systemic infections in humans. These infections mainly originate from the gastrointestinal tract where C. albicans can invade the gut epithelial barrier to gain access to the bloodstream. Along the gut, pathogens can use Microfold (M) cells as a portal of entry to cross the epithelial barrier. M cells are specialized cells mainly located in the follicule-associated epithelium of Peyer patches. In this study, we used scanning electron and fluorescence microscopy, adhesion and invasion assays and fungal mutants to investigate the interactions of C. albicans with M cells obtained in an established in vitro model whereby enterocyte-like Caco-2 cells co-cultured with the Raji B cell line undergo a phenotypic switch to morphologically and functionally resembling M cells. Our data demonstrate that C. albicans co-localizes with and invades preferentially M cells, providing evidence that the fungus can use M cells as a portal of entry into the intestinal barrier. In addition to active penetration, F-actin dependent endocytosis contributes to internalization of the fungus into M cells through a mechanism involving hypha-associated invasins including Ssa1 and Als3.

Published
2016
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2. miR-661 expression in SNAI1-induced epithelial to mesenchymal transition contributes to breast cancer cell invasion by targeting Nectin-1 and StarD10 messengers

Author

Khalil Arar, Christina Laurini, Michèle Sabbah, Laurent Vallar, Michèle Moes, Charles-Henri Lecellier, Evelyne Friederich, A. Le Bechec, Guillaume Vetter, Anne Saumet, Charles Theillet, Le Ster, Yves, Cytoskeleton and Cell Plasticity Lab, Université du Luxembourg (Uni.lu), Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), Université Montpellier 1 (UM1)-CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Centre de Recherche Publique- Santé, Centre de Recherche Saint-Antoine (UMRS893), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Sigma-Aldrich - Evry GENEPOLE, Sigma-Aldrich, Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), This work was supported by grants from the Fond National de la Recherche (FNR) du Luxembourg, (BIOSAN), the Fondation Luxembourgeoise Contre le Cancer, Human Frontier Science Program (RGP0058/2005), INSERM and CNRS, France. A. Saumet is a recipient of a fellowship from the Ministère de la Culture, de l'Enseignement Supérieur et de la Recherche, Luxembourg (BFR 08/046). M. Moes and A. Le Béchec are supported by AFR grants from the Fond National de la Recherche, Luxembourg., CRLCC Val d'Aurelle - Paul Lamarque-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Université du Luxembourg ( Uni.lu ), Institut de recherche en cancérologie de Montpellier ( IRCM ), Université Montpellier 1 ( UM1 ) -CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Montpellier ( UM ), Centre de Recherche Saint-Antoine ( CR Saint-Antoine ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), and Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
SNAI1, Cancer Research, MESH : RNA, Messenger, MESH : Transcription Factors, MESH : Cell Dedifferentiation, Gene Expression, MESH : Breast Neoplasms, Biochemistry, biophysics & molecular biology [F05] [Life sciences], Validation Studies as Topic, MESH : RNA, Small Interfering, medicine.disease_cause, Bioinformatics, MESH : Neoplasm Invasiveness, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, Metastasis, 0302 clinical medicine, MESH : Tumor Cells, Cultured, MESH : Validation Studies as Topic, EMT/miR-661/ SNAI1/StarD10/Nectin-1/breast cancer cell invasion, MESH: RNA, Small Interfering, Tumor Cells, Cultured, MESH : Female, RNA, Small Interfering, Biochimie, biophysique & biologie moléculaire [F05] [Sciences du vivant], Oligonucleotide Array Sequence Analysis, 0303 health sciences, EMT, MESH: Transcription Factors, MESH: Gene Expression Regulation, Neoplastic, MESH : Epithelial Cells, Gene Expression Regulation, Neoplastic, MESH : Oligonucleotide Array Sequence Analysis, MESH: Epithelial Cells, 030220 oncology & carcinogenesis, MESH: Cell Adhesion Molecules, Female, Breast disease, StarD10, Nectin-1, MESH: Gene Expression, MESH : Gene Expression Regulation, Neoplastic, Nectins, Breast Neoplasms, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, MESH: Phosphoproteins, MESH : Cell Adhesion Molecules, MESH: Gene Expression Profiling, 03 medical and health sciences, [SDV.CAN] Life Sciences [q-bio]/Cancer, microRNA, Genetics, medicine, Humans, Gene silencing, Neoplasm Invasiveness, RNA, Messenger, MESH: Tumor Cells, Cultured, Epithelial–mesenchymal transition, Molecular Biology, MESH: RNA, Messenger, 030304 developmental biology, breast cancer cell invasion, MESH: Validation Studies as Topic, MESH: Humans, MESH : Gene Expression Profiling, Gene Expression Profiling, MESH : Humans, Cancer, Epithelial Cells, Mesenchymal Stem Cells, MESH: Neoplasm Invasiveness, Cell Dedifferentiation, Phosphoproteins, medicine.disease, MESH: Cell Dedifferentiation, miR-661, MESH : Gene Expression, MESH : MicroRNAs, MicroRNAs, Retraction Note, MESH: Mesenchymal Stem Cells, MESH: Oligonucleotide Array Sequence Analysis, Cancer research, Snail Family Transcription Factors, MESH : Phosphoproteins, MESH : Mesenchymal Stem Cells, Carcinogenesis, Cell Adhesion Molecules, MESH: MicroRNAs, MESH: Female, MESH: Breast Neoplasms, Transcription Factors
Abstract

4; International audience; Epithelial to mesenchymal transition (EMT) is a key step toward metastasis. MCF7 breast cancer cells conditionally expressing the EMT master regulator SNAI1 were used to identify early expressed microRNAs (miRNAs) and their targets that may contribute to the EMT process. Potential targets of miRNAs were identified by matching lists of in silico predicted targets and of inversely expressed mRNAs. MiRNAs were ranked based on the number of predicted hits, highlighting miR-661, a miRNA with so far no reported role in EMT. MiR-661 was found required for efficient invasion of breast cancer cells by destabilizing two of its predicted mRNA targets, the cell-cell adhesion protein Nectin-1 and the lipid transferase StarD10, resulting, in turn, in the downregulation of epithelial markers. Reexpression of Nectin-1 or StarD10 lacking the 3'-untranslated region counteracted SNAI1-induced invasion. Importantly, analysis of public transcriptomic data from a cohort of 295 well-characterized breast tumor specimen revealed that expression of StarD10 is highly associated with markers of luminal subtypes whereas its loss negatively correlated with the EMT-related, basal-like subtype. Collectively, our non-a priori approach revealed a nonpredicted link between SNAI1-triggered EMT and the down-regulation of Nectin-1 and StarD10 through the up-regulation of miR-661, which may contribute to the invasion of breast cancer cells and poor disease outcome.

Published
2010
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3. Membrane Translocation of Binary Actin-ADP-Ribosylating Toxins from Clostridium difficile and Clostridium perfringens Is Facilitated by Cyclophilin A and Hsp90

Author

Eva Kaiser, Klaus Aktories, Johannes Buchner, Carsten Schwan, Michel R. Popoff, Katharina Ernst, Claudia Kroll, Gunter Fischer, Holger Barth, University of Ulm, University of Freiburg [Freiburg], Bactéries anaérobies et Toxines, Institut Pasteur [Paris], Max-Planck Research Unit for Enzymology of Protein Folding, Max Planck Society (GERMANY), Technische Universität München [München] ( TUM ), This work was financially supported by the Deutsche Forschungsgemeinschaft (grant BA 2087/2-1 to H.B. and grant AK6/20-1 to K.A.), We thank Ulrike Binder for excellent technical assistance., Universität Ulm - Ulm University [Ulm, Allemagne], Max Planck Research Unit for Enzymology of Protein Folding, Max-Planck-Gesellschaft, Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM), and Institut Pasteur [Paris] (IP)

Subjects
MESH: ADP Ribose Transferases, Clostridium perfringens, [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, MESH : Actins, medicine.disease_cause, chemistry.chemical_compound, Cytosol, MESH: Cytosol, MESH : Protein Transport, Chlorocebus aethiops, MESH : Bacterial Proteins, MESH: Animals, MESH : Clostridium difficile, MESH: Bacterial Proteins, Cyclophilin, MESH: Clostridium perfringens, ADP Ribose Transferases, 0303 health sciences, biology, MESH : Cytosol, MESH: Cyclophilin A, MESH : Macrolides, Bafilomycin, MESH : HSP90 Heat-Shock Proteins, Hsp90, Radicicol, MESH : Epithelial Cells, Protein Transport, Infectious Diseases, Biochemistry, MESH: Epithelial Cells, [SDV.TOX]Life Sciences [q-bio]/Toxicology, MESH: Caco-2 Cells, Macrolides, MESH: Macrolides, MESH : ADP Ribose Transferases, Cyclophilin A, MESH : Clostridium perfringens, MESH: Protein Transport, MESH : Cell Membrane, Immunology, MESH: Vero Cells, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH : Cyclophilin A, MESH: Actins, Microbiology, 03 medical and health sciences, Bacterial Proteins, MESH: HSP90 Heat-Shock Proteins, medicine, Animals, Humans, HSP90 Heat-Shock Proteins, Vero Cells, MESH: Clostridium difficile, 030304 developmental biology, MESH: Humans, Cellular Microbiology: Pathogen-Host Cell Molecular Interactions, Clostridioides difficile, 030306 microbiology, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, Cell Membrane, MESH : Humans, MESH : Caco-2 Cells, Epithelial Cells, Pseudomembranous colitis, MESH: Cercopithecus aethiops, Actins, chemistry, biology.protein, Parasitology, MESH : Animals, MESH : Cercopithecus aethiops, Caco-2 Cells, MESH: Cell Membrane, MESH : Vero Cells
Abstract

Some hypervirulent strains of Clostridium difficile produce the binary actin-ADP-ribosylating toxin C. difficile transferase (CDT) in addition to Rho-glucosylating toxins A and B. It has been suggested that the presence of CDT increases the severity of C. difficile -associated diseases, including pseudomembranous colitis. CDT contains a binding and translocation component, CDTb, that mediates the transport of the separate enzyme component CDTa into the cytosol of target cells, where CDTa modifies actin. Here we investigated the mechanism of cellular CDT uptake and found that bafilomycin A1 protects cultured epithelial cells from intoxication with CDT, implying that CDTa is translocated from acidified endosomal vesicles into the cytosol. Consistently, CDTa is translocated across the cytoplasmic membranes into the cytosol when cell-bound CDT is exposed to acidic medium. Radicicol and cyclosporine A, inhibitors of the heat shock protein Hsp90 and cyclophilins, respectively, protected cells from intoxication with CDT but not from intoxication with toxins A and B. Moreover, both inhibitors blocked the pH-dependent membrane translocation of CDTa, strongly suggesting that Hsp90 and cyclophilin are crucial for this process. In contrast, the inhibitors did not interfere with the ADP-ribosyltransferase activity, receptor binding, or endocytosis of the toxin. We obtained comparable results with the closely related iota-toxin from Clostridium perfringens . Moreover, CDTa and Ia, the enzyme component of iota-toxin, specifically bound to immobilized Hsp90 and cyclophilin A in vitro . In combination with our recently obtained data on the C2 toxin from C. botulinum , these results imply a common Hsp90/cyclophilin A-dependent translocation mechanism for the family of binary actin-ADP-ribosylating toxins.

Published
2011
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4. Variable beta-glucans production by different states of Eurotium amstelodami explains differences in inflammatory responses in airway cells

Author

Stéphane Bretagne, Sandrine Roussel, Anne-Pauline Bellanger, Bénédicte Rognon, Laurence Millon, Françoise Botterel, Gabriel Reboux, Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Laboratoire de Parasitologie-Mycologie, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ), Service de parasitologie [Mondor], Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), and Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)

Subjects
MESH: Inflammation, MESH : Cell Line, Chemokine, MESH : Spores, Fungal, beta-Glucans, MESH : beta-Glucans, MESH : Cytokines, MESH: Eurotium, Conidium, Immunology and Allergy, MESH : Fungal Proteins, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, MESH : Eurotium, 0303 health sciences, Fungal protein, MESH: Cytokines, biology, MESH: beta-Glucans, General Medicine, Spores, Fungal, MESH: Chemokines, MESH : Epithelial Cells, MESH : Air Pollution, Indoor, MESH: Epithelial Cells, Air Pollution, Indoor, Cytokines, Proteoglycans, MESH: Fungal Proteins, medicine.symptom, Chemokines, Microbiology (medical), Hypha, Hyphae, Inflammation, Fungus, Pathology and Forensic Medicine, Microbiology, Cell Line, Fungal Proteins, 03 medical and health sciences, MESH: Hyphae, MESH: Spores, Fungal, Eurotium, medicine, Humans, MESH : Chemokines, Interleukin 8, 030304 developmental biology, A549 cell, MESH: Humans, MESH : Inflammation, 030306 microbiology, fungi, MESH : Humans, Epithelial Cells, biology.organism_classification, MESH: Cell Line, Immunology, biology.protein, MESH : Hyphae, MESH: Air Pollution, Indoor
Abstract

International audience; Eurotium amstelodami, a mold frequently identified in housing and farm air samples, is a suspected cause of respiratory diseases such as allergic alveolitis, atopic asthma, and organic dust toxic syndrome. This fungus is present in the air in three different states (ascospores, conidia, and hyphae). The aim of this study was to test in vitro the differential inflammatory response of airway cells exposed to 1,3 betaglucanase-treated protein extract (BGPE), from E. amstelodami ascospores, conidia, and hyphae. Confluent cells from the A549 cell line were inoculated with calibrated BGPE issued from the three fungal forms. The levels of eight cytokines and chemokines involved in inflammatory responses were measured after 8 h of exposure. Beta-d-glucan (BDG) was quantified in total fungal extract as well as in the BGPE from the three fungal states. Hyphal BGPE were the only ones to induce a marked inflammatory response and they contain higher quantities of BDG. The present study adds to the growing body of evidence that beta-glucan from fungal hyphae play a crucial role in respiratory diseases.

Published
2011
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5. Cellular transcriptional profiling in human lung epithelial cells infected by different subtypes of influenza A viruses reveals an overall down-regulation of the host p53 pathway

Author

Catherine Nguyen, Laurence Josset, Jean-Christophe Bourdon, Bruno Lina, Jean-Jacques Diaz, Olivier Ferraris, Virginie Marcel, Olivier Terrier, Julien Textoris, Manuel Rosa-Calatrava, Gaëlle Cartet, Division of Medical Sciences, University of Dundee-Centre for Oncology and Molecular Medicine, Virologie et pathologie humaine ( VirPath ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon, Laboratoire de Virologie Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon ( HCL ), Technologies avancées pour le génôme et la clinique ( TAGC ), Aix Marseille Université ( AMU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Centre de génétique et de physiologie moléculaire et cellulaire ( CGPhiMC ), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique ( CNRS ), This work was supported by University Claude Bernard of Lyon, the Civil hospitals of Lyon, the AVIESAN alliance and by a grant of the 'Programme de Recherche A (H1N1)' co-ordinated by the Institut de Microbiologie et Maladies Infectieuses (INSERM)., Virologie et pathologie humaine (VirPath), Université Claude Bernard Lyon 1 (UCBL), Hospices Civils de Lyon (HCL), Technologies avancées pour le génôme et la clinique (TAGC), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de génétique et de physiologie moléculaire et cellulaire (CGPhiMC), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), and BMC, Ed.

Subjects
MESH: Signal Transduction, MESH : Cell Line, MESH : Blotting, Western, medicine.disease_cause, MESH: Down-Regulation, Transcriptome, MESH : Down-Regulation, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Reverse Transcriptase Polymerase Chain Reaction, Gene expression, Influenza A virus, MESH : Influenza, Human, MESH: Tumor Suppressor Protein p53, 0303 health sciences, Reverse Transcriptase Polymerase Chain Reaction, MESH: Influenza, Human, MESH : Reverse Transcriptase Polymerase Chain Reaction, 3. Good health, MESH : Epithelial Cells, MESH : Influenza A virus, [ SDV.MHEP.MI ] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Infectious Diseases, MESH: Epithelial Cells, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Signal Transduction, Blotting, Western, MESH: Influenza A virus, Down-Regulation, Biology, Cell Line, lcsh:Infectious and parasitic diseases, 03 medical and health sciences, MESH: Gene Expression Profiling, Virology, Influenza, Human, medicine, Humans, MESH: Blotting, Western, lcsh:RC109-216, Gene, 030304 developmental biology, MESH : Signal Transduction, MESH: Humans, 030306 microbiology, Gene Expression Profiling, Research, MESH : Gene Expression Profiling, MESH : Humans, Epithelial Cells, Influenza A virus subtype H5N1, MESH: Cell Line, Gene expression profiling, MESH : Tumor Suppressor Protein p53, Viral replication, Cell culture, Tumor Suppressor Protein p53
Abstract

BackgroundInfluenza viruses can modulate and hijack several cellular signalling pathways to efficiently support their replication. We recently investigated and compared the cellular gene expression profiles of human lung A549 cells infected by five different subtypes of human and avian influenza viruses (Jossetet al.Plos One 2010). Using these transcriptomic data, we have focused our analysis on the modulation of the p53 pathway in response to influenza infection.ResultsOur results were supported by both RT-qPCR and western blot analyses and reveal multiple alterations of the p53 pathway during infection. A down-regulation of mRNA expression was observed for the main regulators of p53 protein stability during infection by the complete set of viruses tested, and a significant decrease in p53 mRNA expression was also observed in H5N1 infected cells. In addition, several p53 target genes were also down-regulated by these influenza viruses and the expression of their product reduced.ConclusionsOur data reveal that influenza viruses cause an overall down-regulation of the host p53 pathway and highlight this pathway and p53 protein itself as important viral targets in the altering of apoptotic processes and in cell-cycle regulation.

Published
2011
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6. New evidence of the involvement of Lichtheimia corymbifera in farmer's lung disease

Author

Bénédicte Rognon, Stéphane Bretagne, Sandrine Roussel, Jean-Charles Dalphin, Anne-Pauline Bellanger, Laurence Millon, Charline Candido, Gabriel Reboux, Françoise Botterel, Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Parasitologie-Mycologie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Service de parasitologie [Mondor], Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), and Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 )

Subjects
MESH: Inflammation, MESH: Interleukin-13, Allergy, MESH : RNA, Messenger, Lichtheimia corymbifera, Gene Expression, MESH : Farmer's Lung, 0302 clinical medicine, Absidia, MESH: Up-Regulation, MESH : Up-Regulation, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, 0303 health sciences, Interleukin-13, biology, Farmer's lung, General Medicine, Precipitin, MESH : Epithelial Cells, 3. Good health, Up-Regulation, Infectious Diseases, MESH : Mucorales, MESH: Epithelial Cells, MESH: Mucorales, Mucorales, Hypersensitivity pneumonitis, MESH : Interleukin-8, MESH: Cell Line, Tumor, MESH: Gene Expression, Microbiology, 03 medical and health sciences, Cell Line, Tumor, MESH : Interleukin-13, medicine, Humans, Mucormycosis, MESH : Mucormycosis, RNA, Messenger, MESH: RNA, Messenger, MESH: Mucormycosis, Pneumonitis, MESH: Farmer's Lung, Inflammation, MESH : Inflammation, MESH: Humans, MESH : Cell Line, Tumor, 030306 microbiology, MESH : Humans, fungi, Interleukin-8, Epithelial Cells, biology.organism_classification, medicine.disease, MESH: Interleukin-8, Spore, MESH : Gene Expression, 030228 respiratory system, Immunology, Farmer's Lung
Abstract

International audience; Farmer's lung disease (FLD) is a form of hypersensitivity pneumonitis resulting from recurrent exposure to moldy plant materials. We investigated and compared the initial response of respiratory epithelium after exposure to extracts of Sacharopolyspora rectivirgula, Lichtheimia corymbifera (formerly Absidia corymbifera), Eurotium amstelodami and Wallemia sebi. The two criteria for selection of these species were their high prevalence in the hay handled by FLD patients and the presence of high levels of specific precipitins to these molds in FLD patients' sera. Hydrosoluble extracts were prepared from spores and hyphae grown in culture under optimal conditions for each of the four species. Confluent A549 cells were inoculated with one of the four calibrated soluble extracts. Two mediators, one inflammatory (Interleukin (IL)-8) and one allergic (IL-13), were quantified using real-time PCR and ELISA assay, after four exposure periods (30 min, 2 h, 4 h and 8 h). S. rectivirgula and L. corymbifera extracts were the only ones which induced a marked upregulation of IL-8, as shown by both real-time PCR and ELISA assay 8 h after the initial contact. This study adds to the growing body of evidence that L. corymbifera should be recognized as an etiologic agent of FLD along with S. rectivirgula.

Published
2010
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7. A capsid-encoded PPxY-motif facilitates adenovirus entry

Author

Baptist Jammart, Zsolt Ruzsics, Eric J. Kremer, Daniel Henaff, Sigrid Seelmeir, Harald Wodrich, Carolina Segura-Morales, Christopher M. Wiethoff, Olivier Coux, Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Max Von Pettenkofer Institute (MVP), Ludwig-Maximilians-Universität München (LMU), Centre de recherche en Biologie Cellulaire (CRBM), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Loyola University Medical Center (LUMC), Microbiologie cellulaire et moléculaire et pathogénicité ( MCMP ), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique ( CNRS ), Institut de Recherche en Infectiologie de Montpellier ( IRIM ), Centre National de la Recherche Scientifique ( CNRS ) -Université de Montpellier ( UM ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Max Von Pettenkofer Institute ( MVP ), Ludwig-Maximilians-Universität München, Centre de recherches de biochimie macromoléculaire ( CRBM ), Université Montpellier 1 ( UM1 ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -IFR122-Centre National de la Recherche Scientifique ( CNRS ), Loyola University Medical Center ( LUMC ), Automatic mesh generation and advanced methods ( Gamma3 ), Institut Charles Delaunay ( ICD ), Université de Technologie de Troyes ( UTT ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Technologie de Troyes ( UTT ) -Centre National de la Recherche Scientifique ( CNRS ) -Inria Saclay - Ile de France, and Institut National de Recherche en Informatique et en Automatique ( Inria ) -Institut National de Recherche en Informatique et en Automatique ( Inria )

Subjects
MESH: Osteosarcoma, Nedd4 Ubiquitin Protein Ligases, MESH : Capsid Proteins, MESH: Microtubule-Organizing Center, NEDD4, Retinal Pigment Epithelium, Microtubules, MESH : Ubiquitination, Conserved sequence, MESH: Endosomal Sorting Complexes Required for Transport, Adenovirus Infections, Human, MESH: Protein Structure, Tertiary, Ubiquitin, [ SDV.MP ] Life Sciences [q-bio]/Microbiology and Parasitology, Biology (General), Internalization, Virology/Virion Structure, Assembly, and Egress, Lung, Conserved Sequence, MESH : Microtubules, MESH: Capsid Proteins, media_common, MESH : Adenoviruses, Human, 0303 health sciences, Osteosarcoma, MESH : Osteosarcoma, MESH: Conserved Sequence, MESH: Microtubules, 030302 biochemistry & molecular biology, MESH : Adenovirus Infections, Human, MESH: Retinal Pigment Epithelium, 3. Good health, Cell biology, MESH : Epithelial Cells, MESH : Microtubule-Organizing Center, MESH: Adenovirus Infections, Human, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Capsid, MESH : Retinal Pigment Epithelium, MESH: Epithelial Cells, MESH : Protein Structure, Tertiary, Research Article, MESH: Adenoviruses, Human, MESH: Cell Line, Tumor, Endosome, QH301-705.5, media_common.quotation_subject, Ubiquitin-Protein Ligases, MESH : Lung, Immunology, Endosomes, Biology, Microbiology, 03 medical and health sciences, Viral entry, MESH : Conserved Sequence, Virology, Cell Line, Tumor, Genetics, Humans, MESH: Lung, Molecular Biology, 030304 developmental biology, MESH: Humans, Endosomal Sorting Complexes Required for Transport, MESH : Cell Line, Tumor, Adenoviruses, Human, MESH : Humans, Ubiquitination, Epithelial Cells, RC581-607, Virology/Host Invasion and Cell Entry, MESH: Ubiquitin-Protein Ligases, Protein Structure, Tertiary, MESH: Endosomes, biology.protein, MESH : Endosomal Sorting Complexes Required for Transport, MESH : Endosomes, MESH: Ubiquitination, Parasitology, Capsid Proteins, MESH : Ubiquitin-Protein Ligases, Immunologic diseases. Allergy, Microtubule-Organizing Center
Abstract

Viruses use cellular machinery to enter and infect cells. In this study we address the cell entry mechanisms of nonenveloped adenoviruses (Ads). We show that protein VI, an internal capsid protein, is rapidly exposed after cell surface attachment and internalization and remains partially associated with the capsid during intracellular transport. We found that a PPxY motif within protein VI recruits Nedd4 E3 ubiquitin ligases to bind and ubiquitylate protein VI. We further show that this PPxY motif is involved in rapid, microtubule-dependent intracellular movement of protein VI. Ads with a mutated PPxY motif can efficiently escape endosomes but are defective in microtubule-dependent trafficking toward the nucleus. Likewise, depletion of Nedd4 ligases attenuates nuclear accumulation of incoming Ad particles and infection. Our data provide the first evidence that virus-encoded PPxY motifs are required during virus entry, which may be of significance for several other pathogens., Author Summary Viruses exploit cellular functions during entry and exit of cells. To redirect cellular functions for their own purpose, viruses encode high-affinity binding sites for key-cellular factors. One such domain is the PPxY motif, which is present in structural proteins of several, mainly enveloped viruses. This motif binds to ubiquitin ligases of the Nedd4 family and recruits their function to sites of virus budding from cells. Here we show that adenoviruses also encode a PPxY motif in the internal structural protein VI and that the PPxY motif has an unprecedented function in virus entry. Adenoviruses with mutations in the protein VI PPxY motif undergo normal endosomal uptake and membrane penetration but have reduced infectivity, altered intracellular targeting and lack efficient gene-delivery. We also find that protein VI is ubiquitylated by Nedd4 ligases in a PPxY dependent manner following partial capsid disassembly and displays rapid intracellular movement. Depletion of Nedd4 ligases also alters virus movement within cells during entry and reduces viral infectivity. Given that PPxY motifs are important for virus exit our findings might have uncovered an additional function for PPxY motifs in virus entry, potentially expanding the significance of PPxY motifs and functionally related domains for viral replication.

Published
2010
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8. Muco-ciliary differentiation of nasal epithelial cells is decreased after wound healing in vitro. : Muco-ciliary differentiation & wound healing

Author

Lazard, Diane, Moore, Anne, Hupertan, Vincent, Martin, Clémence, Escabasse, Virginie, Dreyfus, Patrick, Burgel, Pierre-Régis, Amselem, Serge, Escudier, Estelle, Coste, André, INSERM U955, équipe 13, Institut Mondor de Recherche Biomédicale ( IMRB ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ), Physiopathologie des maladies génétiques d'expression pédiatrique, Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ), Statistics and data mining, Service de pneumologie [CHU Cochin], CHU Cochin [AP-HP]-Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP]-Assistance publique - Hôpitaux de Paris (AP-HP)-Institut Mondor de Recherche Biomédicale ( IMRB ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ) -Mucoviscidose et bronchopathies chroniques : biopathologie et phénotypes cliniques ( EA 2511 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Université Paris Descartes - Paris 5 ( UPD5 ), Service d'ORL et de Chirurgie Cervico-Faciale, CHI Créteil-CHI Créteil-Service d'ORL et de chirurgie cervico-faciale, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Institut Mondor de Recherche Biomédicale ( IMRB ), Thérapie des maladies du muscle strié, Centre National de la Recherche Scientifique ( CNRS ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ), Mucoviscidose et bronchopathies chroniques : biopathologie et phénotypes cliniques ( EA 2511 ), Université Paris Descartes - Paris 5 ( UPD5 ), CHU Cochin [AP-HP]-Assistance publique - Hôpitaux de Paris (AP-HP), This work was supported by grants from the Legs Poix from the Chancellerie des Universités, Dyskinésie ciliaire primitive : aspects génétiques et fonctionnels, PCD Project, ANR-05-MRAR-022-01,Dyskinésie ciliaire primitive : aspects génétiques et fonctionnels, PCD Project, ANR-05-MRAR-022-01, Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Mucoviscidose et bronchopathies chroniques : biopathologie et phénotypes cliniques (EA 2511), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Institut Mondor de Recherche Biomédicale (IMRB), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Mucoviscidose et bronchopathies chroniques : biopathologie et phénotypes cliniques (EA 2511), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Descartes - Paris 5 (UPD5), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), ANR-05-MRAR-0022,PCD Project,Dyskinésie ciliaire primitive : aspects génétiques et fonctionnels(2005), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-IFR10-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-IFR10, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-IFR10-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-IFR10-Mucoviscidose et bronchopathies chroniques : biopathologie et phénotypes cliniques (EA 2511), and Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)

Subjects
MESH: Cell Differentiation, ciliated cells, MESH : Dyneins, MESH : Mucin 5AC, MESH : Wound Healing, wound healing, MESH: Tubulin, MESH: Mucins, MESH: Down-Regulation, mucous cells, MESH : Down-Regulation, MESH: Transforming Growth Factor beta1, MESH : Cilia, MESH: Cilia, MESH : Keratin-14, MESH : Cells, Cultured, [SDV.GEN]Life Sciences [q-bio]/Genetics, MESH: Humans, MESH: Mucin 5AC, MESH : Humans, respiratory system, MESH : Tubulin, MESH : Axonemal Dyneins, MESH : Epithelial Cells, MESH : Nasal Mucosa, MESH: Wound Healing, human nasal epithelial differentiation, MESH : Nasal Polyps, MESH: Nasal Polyps, MESH: Epithelial Cells, MESH: Nasal Mucosa, MESH : Mucins, MESH: Keratin-14, MESH: Axonemal Dyneins, MESH: Dyneins, MESH : Transforming Growth Factor beta1, [ SDV.GEN ] Life Sciences [q-bio]/Genetics, MESH : Cell Differentiation, TGF-beta 1, MESH: Cells, Cultured
Abstract

International audience; BACKGROUND: Epithelial damage and modifications of cell differentiation are frequent in airway diseases with chronic inflammation, in which transforming growth factor-beta1 (TGF-beta1) plays an important role. The aim of this study was to evaluate the differentiation of human nasal epithelial cells (HNEC) after wound healing and the potential effects of TGF-beta1. METHODS: Basal, mucus, and ciliated cells were characterized by cytokeratin-14, MUC5AC, and betaIV tubulin immunodetection, respectively. Their expression was evaluated in situ in nasal polyps and in an in vitro model of wound healing in primary cultures of HNEC after wound closure, under basal conditions and after TGF-beta1 supplementation. Using RT-PCR, the effects of TGF-beta1 on MUC5AC and DNAI1 genes, specifically transcribed in mucus and ciliated cells, were evaluated. RESULTS: In situ, high TGF-beta1 expression was associated with low MUC5AC and betaIV tubulin expression. In vitro, under basal conditions, MUC5AC expression remained stable, cytokeratin-14 expression was strong and decreased with time, while betaIV tubulin expression increased. Transforming growth factor-beta1 supplementation downregulated MUC5AC and betaIV tubulin expression as well as MUC5AC and DNAI1 transcripts. CONCLUSION: After a wound, differentiation into mucus and ciliated cells was possible and partially inhibited in vitro by TGF-beta1, a cytokine that may be involved in epithelial remodeling observed in chronic airway diseases.

Published
2009
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9. Aspergillus fumigatus germ tube growth and not conidia ingestion induces expression of inflammatory mediator genes in the human lung epithelial cell line A549

Author

Stéphane Bretagne, Anne-Pauline Bellanger, Ingrid Laurendeau, Laurence Millon, Ivan Bièche, Françoise Botterel, D Rivollet, Michel Vidaud, Khaled Khoufache, Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Parasitologie-Mycologie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Genetique Moleculaire des Cancers d'Origine Epitheliale, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie [Paris], Service de parasitologie [Mondor], Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ), and Institut National de la Santé et de la Recherche Médicale ( INSERM ) -INSTITUT CURIE

Subjects
MESH : Cell Line, MESH : Spores, Fungal, Necrosis, MESH : Respiratory Mucosa, MESH : Cytokines, Penicillium chrysogenum, Polymerase Chain Reaction, Aspergillus fumigatus, MESH: Respiratory Mucosa, MESH : Immunosuppressive Agents, skin and connective tissue diseases, MESH : Polymerase Chain Reaction, [ SDV.MP.MYC ] Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, 0303 health sciences, MESH: Cytokines, biology, Interleukin, General Medicine, Spores, Fungal, MESH : Epithelial Cells, MESH: Epithelial Cells, Cytokines, Steroids, Tumor necrosis factor alpha, MESH: Immunosuppressive Agents, medicine.symptom, MESH: Aspergillus fumigatus, MESH: Penicillium chrysogenum, Immunosuppressive Agents, Microbiology (medical), MESH : Penicillium chrysogenum, Inflammation, Respiratory Mucosa, Microbiology, Cell Line, 03 medical and health sciences, MESH: Gene Expression Profiling, MESH: Spores, Fungal, medicine, Humans, Interleukin 8, 030304 developmental biology, A549 cell, MESH: Humans, 030306 microbiology, Gene Expression Profiling, MESH : Gene Expression Profiling, MESH : Humans, Epithelial Cells, MESH: Polymerase Chain Reaction, biology.organism_classification, MESH: Steroids, MESH: Cell Line, MESH : Aspergillus fumigatus, Respiratory epithelium, MESH : Steroids
Abstract

Inhalation of conidia is the main cause of invasive pulmonary aspergillosis (IPA) and the respiratory epithelium is the first line of defence. To explore the triggering factor for the inflammatory response toAspergillus fumigatus, the species mainly responsible for IPA, this study analysed the differential expression of three inflammatory genes in A549 cells after challenge with live and killed conidia. The influence of steroids, one of the main risk factors for developing IPA, was also investigated. Quantification of mRNAs of the inflammatory mediator genes encoding interleukin (IL)-8, tumour necrosis factor (TNF)-αand granulocyte–monocyte colony-stimulating factor (GM-CSF) was carried out using real-time PCR. Ingestion rates were studied for the conidia ofA. fumigatusandPenicillium chrysogenumusing a fluorescence brightener. Similar results were obtained for both species, with ingestion rates ranging from 35 to 40 %. Exposure of A549 cells to liveA. fumigatusconidia only induced a four- to fivefold increase in the mRNA levels of the three genes, starting 8 h after the initial contact. Both inactivation of liveA. fumigatusconidia and treatment by dexamethasone (10−7 M) prevented the overexpression of TNF-α, IL-8 and GM-CSF. Fungal growth, rather than conidia ingestion, appears to be the main stimulus for the production of inflammatory mediators by epithelial cells, and this production is inhibited by steroid therapy. These results underline the role that the epithelium plays in the innate response against IPA.

Published
2009
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10. Cell-cell fusion induced by measles virus amplifies the type I interferon response

Author

Johan Druelle, Timothy Fabian Wild, Florence Herschke, Denis Gerlier, Sébastien Plumet, Chantal Rabourdin-Combe, David Laine, Hélène Valentin, Olga Azocar, Thomas Duhen, Virologie et Pathologie Humaine (VirPath), École normale supérieure - Lyon (ENS Lyon)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Immunobiologie fondamentale et clinique, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), BioSciences Lyon-Gerland (BLG), École normale supérieure - Lyon (ENS Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Virologie humaine, École normale supérieure - Lyon (ENS Lyon)-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported in part by grants from ANR (DG, ANR-MIME), INSERM, INCA-Canceropole 2004-2005 and ARC 3637. D.L., F.H., J.D., T.D., and S.P. were supported by a fellowship from MENRT, ARC and DGA, respectively., Measles and Interferon response, École normale supérieure de Lyon (ENS de Lyon)-Université Claude Bernard Lyon 1 (UCBL), École normale supérieure de Lyon (ENS de Lyon)-Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), École normale supérieure de Lyon (ENS de Lyon)-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), Virologie et Pathologie Humaine ( VirPath ), École normale supérieure - Lyon ( ENS Lyon ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-IFR128-Institut National de la Santé et de la Recherche Médicale ( INSERM ), BioSciences Lyon-Gerland ( BLG ), École normale supérieure - Lyon ( ENS Lyon ) -Institut National de la Recherche Agronomique ( INRA ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Hospices Civils de Lyon ( HCL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), École normale supérieure - Lyon ( ENS Lyon ) -IFR128-Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Gerlier, Denis

Subjects
MESH : Cell Line, MESH: Interferon Type I, fusion, Interferon Regulatory Factor-7, MESH : Viral Fusion Proteins, Cellular Response to Infection, Giant Cells, MESH: Interferon Regulatory Factor-7, MESH : Virology, Cell Fusion, MESH: Giant Cells, MESH : Interferon Regulatory Factor-7, MESH: Interferon Regulatory Factor-3, Interferon, Chlorocebus aethiops, MESH : Interferon Regulatory Factor-3, MESH : Innate immunity, MESH: Animals, MESH : Viral Proteins, Mononegavirales, OCIS 000.1430, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH : Cell Nucleus, Cell fusion, Microscopy, Video, biology, MESH: Dendritic Cells, MESH : Microscopy, Video, interferon, MESH: Innate immunity, MESH : Epithelial Cells, MESH: Epithelial Cells, Interferon Type I, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Viral Fusion Proteins, medicine.drug, MESH: Cell Nucleus, Immunology, Microbiology, [ SDV.MP.VIR ] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Virus, Cell Line, Measles virus, Viral Proteins, MESH : Interferon Type I, Virology, medicine, Animals, Humans, dendritic cells, MESH: Immunology, Cell Nucleus, MESH: Humans, MESH : Cell Fusion, MESH : Humans, MESH : Giant Cells, biology.organism_classification, Molecular biology, MESH: Viral Proteins, MESH: Cercopithecus aethiops, epithelial cells, MESH: Cell Line, MESH: Microscopy, Video, Giant cell, Cell culture, Insect Science, MESH : Dendritic Cells, measles virus, MESH: Cell Fusion, MESH : Immunology, MESH : Measles virus, Interferon Regulatory Factor-3, MESH : Animals, MESH : Cercopithecus aethiops, Viral Fusion Proteins, MESH: Measles virus, Interferon type I, MESH: Virology
Abstract

Measles virus (MeV) infection is characterized by the formation of multinuclear giant cells (MGC). We report that beta interferon (IFN-β) production is amplified in vitro by the formation of virus-induced MGC derived from human epithelial cells or mature conventional dendritic cells. Both fusion and IFN-β response amplification were inhibited in a dose-dependent way by a fusion-inhibitory peptide after MeV infection of epithelial cells. This effect was observed at both low and high multiplicities of infection. While in the absence of virus replication, the cell-cell fusion mediated by MeV H/F glycoproteins did not activate any IFN-α/β production, an amplified IFN-β response was observed when H/F-induced MGC were infected with a nonfusogenic recombinant chimerical virus. Time lapse microscopy studies revealed that MeV-infected MGC from epithelial cells have a highly dynamic behavior and an unexpected long life span. Following cell-cell fusion, both of the RIG-I and IFN-β gene deficiencies were trans complemented to induce IFN-β production. Production of IFN-β and IFN-α was also observed in MeV-infected immature dendritic cells (iDC) and mature dendritic cells (mDC). In contrast to iDC, MeV infection of mDC induced MGC, which produced enhanced amounts of IFN-α/β. The amplification of IFN-β production was associated with a sustained nuclear localization of IFN regulatory factor 3 (IRF-3) in MeV-induced MGC derived from both epithelial cells and mDC, while the IRF-7 up-regulation was poorly sensitive to the fusion process. Therefore, MeV-induced cell-cell fusion amplifies IFN-α/β production in infected cells, and this indicates that MGC contribute to the antiviral immune response.

Published
2007
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11. Modulation of p53 transcriptional activity by PRIMA-1 and Pifithrin-alpha on staurosporine-induced apoptosis of wild-type and mutated p53 epithelial cells

Author

Jean-Luc Prétet, J. F. Charlot, Christiane Mougin, Magali Nicolier, Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC ( CEF2P / CARCINO ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC (EA 3181) (CEF2P / CARCINO), Université de Franche-Comté (UFC), and Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)

Subjects
Transcription, Genetic, MESH : Hela Cells, Cell, Mutant, Apoptosis, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, HeLa, 0302 clinical medicine, MESH: Nerve Tissue Proteins, MESH : Cell Transformation, Viral, Enzyme Inhibitors, MESH: Tumor Suppressor Protein p53, 0303 health sciences, MESH : Cell Line, Transformed, DNA, Neoplasm, Pifithrin, Cell biology, Drug Combinations, MESH : Drug Combinations, MESH: Epithelial Cells, 030220 oncology & carcinogenesis, DNA fragmentation, MESH: Membrane Proteins, MESH : Cell Culture Techniques, MESH: Mitochondria, MESH: Toluene, 03 medical and health sciences, Humans, MESH: Membrane Potentials, MESH: Benzothiazoles, MESH: Drug Combinations, Pharmacology, MESH: Cell Culture Techniques, MESH: Humans, MESH : bcl-2-Associated X Protein, MESH : Humans, Epithelial Cells, Staurosporine, Thiazoles, chemistry, MESH : Membrane Proteins, Mutation, MESH: Female, MESH : Apoptosis, HeLa Cells, Cancer Research, MESH : Staurosporine, Clinical Biochemistry, Cell Culture Techniques, Pharmaceutical Science, MESH : Benzothiazoles, Membrane Potentials, chemistry.chemical_compound, MESH: Papillomaviridae, MESH : Thiazoles, MESH : Membrane Potentials, MESH : Female, Papillomaviridae, MESH : Nerve Tissue Proteins, MESH : Toluene, Cell Line, Transformed, bcl-2-Associated X Protein, Mitochondria, MESH : Epithelial Cells, medicine.anatomical_structure, MESH: Cell Transformation, Viral, MESH: Enzyme Inhibitors, Female, MESH : Mitochondria, MESH : DNA, Neoplasm, MESH : Mutation, medicine.drug, MESH: Mutation, MESH: Cell Line, Tumor, MESH: Thiazoles, MESH: DNA, Neoplasm, Nerve Tissue Proteins, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, Cell Line, Tumor, medicine, MESH: bcl-2-Associated X Protein, Benzothiazoles, MESH: Cell Line, Transformed, MESH : Papillomaviridae, 030304 developmental biology, MESH : Enzyme Inhibitors, MESH : Cell Line, Tumor, MESH: Apoptosis, MESH: Transcription, Genetic, Biochemistry (medical), Wild type, Membrane Proteins, MESH : Transcription, Genetic, Cell Biology, Cell Transformation, Viral, biology.organism_classification, Molecular biology, MESH: Hela Cells, MESH : Tumor Suppressor Protein p53, MESH: Staurosporine, Tumor Suppressor Protein p53, Toluene
Abstract

International audience; We recently argued for a major role of p53 in staurosporine(ST)-induced apoptosis of immortalized epithelial cells, depending on their p53 status. Here, we studied the effects of PRIMA-1 (p53 reactivation and induction of massive apoptosis) and Pifithrin-alpha (p fifty-three inhibitor) in combination with ST to reinforce our previous results by respectively restoring or inhibiting the p53 transcriptional activity in different cell lines.PRIMA-1 does modify neither expression of apoptosis-related proteins nor the percentage of wild-type p53 HeLa and CaSki cells with [symbol: see text]delta psi m and DNA cleavage, whilst it increases by 45% Bax expression and apoptosis of mutated p53 C33A cells. Pifithrin-alpha, does modify neither Bax expression nor apoptosis level of C33A cells, but readily inhibits both [symbol: see text]delta psi m and DNA fragmentation of p53wt cells with decreasing Bax expression. These data support the evidence that PRIMA-1 could be a good candidate, as an anti-cancer drug targeting mutant p53, in order to increase ST efficiency. Moreover, Pifithrin-alpha could be used in combination with ST and PRIMA-1 to prevent side effects of anti-tumor therapies in cells expressing mutant P53.

Published
2006
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12. The SrtA Sortase of Streptococcus agalactiae is required for cell wall anchoring of proteins containing the LPXTG motif, for adhesion to epithelial cells, and for colonization of the mouse intestine

Author

Marina Baptista, Christophe Rusniok, Shaynoor Dramsi, Florence Doucet-Populaire, Lila Lalioui, Frank Kunst, Claire Poyart, Patrick Trieu-Cuot, Nadege Bourgeois, Mohamed Zouine, Philippe Glaser, Elisabeth Pellegrini, Pathogénie des infections systémiques (UMR_S 570), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Biologie des Bactéries Pathogènes à Gram-positif, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Microbiologie, Université Paris Descartes - Paris 5 (UPD5), Génomique des Microorganismes Pathogènes, Pathogénie des infections systémiques ( UMR_S 570 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Université Paris Descartes - Paris 5 ( UPD5 ), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH : Molecular Sequence Data, MESH : Aminoacyltransferases, Amino Acid Motifs, Mutant, MESH: Amino Acid Sequence, MESH: Rats, Sprague-Dawley, [ SDV.MP.BAC ] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, medicine.disease_cause, Bacterial Adhesion, Rats, Sprague-Dawley, Mice, chemistry.chemical_compound, C5a peptidase, MESH: Amino Acid Motifs, Cell Wall, Sortase, MESH: Fibronectins, MESH : Fibronectins, MESH : Bacterial Proteins, MESH: Animals, Peptide sequence, MESH: Bacterial Proteins, Cells, Cultured, MESH : Intestines, Mice, Inbred C3H, 0303 health sciences, biology, MESH : Rats, MESH : Amino Acid Sequence, MESH : Streptococcus agalactiae, Aminoacyltransferases, 3. Good health, MESH : Epithelial Cells, Intestines, Cysteine Endopeptidases, Infectious Diseases, MESH: Epithelial Cells, MESH : Cysteine Endopeptidases, MESH: Intestines, MESH: Cells, Cultured, MESH : Amino Acid Motifs, MESH: Rats, MESH : Bacterial Adhesion, Molecular Sequence Data, Immunology, Virulence, Microbiology, Streptococcus agalactiae, 03 medical and health sciences, MESH: Cell Wall, Bacterial Proteins, MESH : Mice, Inbred C3H, MESH: Aminoacyltransferases, MESH : Mice, MESH : Cells, Cultured, medicine, Animals, Humans, Amino Acid Sequence, MESH: Bacterial Adhesion, MESH: Mice, Inbred C3H, MESH: Mice, 030304 developmental biology, MESH : Cell Wall, MESH: Humans, MESH: Molecular Sequence Data, 030306 microbiology, MESH : Humans, Epithelial Cells, Molecular Pathogenesis, MESH: Streptococcus agalactiae, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH : Rats, Sprague-Dawley, Fibronectins, Rats, Fibronectin, chemistry, biology.protein, Parasitology, Peptidoglycan, MESH : Animals, MESH: Cysteine Endopeptidases
Abstract

Streptococcus agalactiae (group B streptococcus [GBS]) is the leading cause of neonatal pneumonia, sepsis, and meningitis. An in silico genome analysis indicated that GBS strain NEM316 encodes 35 proteins containing an LPXTG motif which are thought to be covalently linked to the peptidoglycan by an enzyme called sortase. The role of these cell wall-anchored proteins in GBS pathogenesis was evaluated on a global level by inactivating the srtA gene. This gene encodes the major sortase SrtA that anchors most of the LPXTG-containing proteins. We chose the C5a peptidase (ScpB) and Alp2, an abundant immunogenic protein, as prototypical LPXTG-containing proteins. As expected, the SrtA knockout mutant was unable to anchor the C5a peptidase (ScpB) and Alp2 to the cell wall. Complementation with plasmid-borne srtA inserted into the chromosome restored the correct surface localization of both ScpB and Alp2. Interestingly, the SrtA mutant was impaired for binding to the major extracellular matrix components fibronectin and fibrinogen and displayed a significant reduction in adherence to human (A549, HeLa, and Caco-2) and murine (L2) epithelial cells compared to the parental wild-type strain. Surprisingly, the inactivation of srtA had no effect on the virulence of the type III strain of GBS in a neonatal rat model (measured by the 50% lethal dose and lung colonization) but strongly impaired the capacity of the strain to colonize the intestines of gnotobiotic mice in a competition assay. These results demonstrate that LPXTG-containing proteins are involved in cell adhesion and GBS persistence in vivo.

Published
2005
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13. CovS/CovR of group B streptococcus: a two-component global regulatory system involved in virulence

Author

Marie-Cécile, Lamy, Mohammed, Zouine, Juliette, Fert, Massimo, Vergassola, Elisabeth, Couve, Elisabeth, Pellegrini, Philippe, Glaser, Frank, Kunst, Tarek, Msadek, Patrick, Trieu-Cuot, Claire, Poyart, Biologie des Bactéries Pathogènes à Gram-positif, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Pathogénie des infections systémiques ( UMR_S 570 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Bactériologie, CHU Cochin [AP-HP], Génomique des Microorganismes Pathogènes, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Pathogénie des infections systémiques (UMR_S 570), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), This work was supported by research funds from the Institut National de la Santé et de la Recherche Médicale, the European Commission (Grant QLG2‐CT‐1999‐01455), the Centre National de la Recherche Scientifique, the Institut Pasteur (PTR No. 17 and GPH No. 9), and the Universities Paris 5 and Paris 7. M.‐C. Lamy was supported by the Ministère de la Recherche et des Technologies and the Fondation pour la Recherche Médicale., We are grateful to J.L. Berretti for determination of haemolytic activities, D. Euphrasie for help in the construction of the ΔCylE mutant, and S. Dubrac and C. Buchrieser for helpful discussion., and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Signal Transduction, MESH : Virulence Factors, MESH : Hemolysis, Transcription, Genetic, MESH : Operon, MESH : Streptococcal Infections, MESH : Gene Deletion, MESH: Virulence, [ SDV.MP.BAC ] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Bacterial Adhesion, Hemolysin Proteins, MESH: Streptococcal Infections, MESH : Bacterial Proteins, MESH: Animals, Promoter Regions, Genetic, MESH: Bacterial Proteins, MESH: Gene Expression Regulation, Bacterial, Virulence, MESH : Rats, MESH : Genes, Bacterial, MESH : Virulence, MESH : Streptococcus agalactiae, MESH: Hemolysis, MESH : Epithelial Cells, MESH: Promoter Regions (Genetics), MESH: Hemolysin Proteins, MESH: Repressor Proteins, MESH: Epithelial Cells, MESH: Regulon, MESH: Genes, Bacterial, MESH : Repressor Proteins, Signal Transduction, MESH : Gene Expression Regulation, Bacterial, MESH: Operon, MESH: Rats, Virulence Factors, MESH : Bacterial Adhesion, MESH : Regulon, MESH : Hemolysin Proteins, MESH : Protein Kinases, Hemolysis, Regulon, Streptococcus agalactiae, Lethal Dose 50, MESH: Gene Expression Profiling, Bacterial Proteins, Streptococcal Infections, Operon, Animals, Humans, MESH : Lethal Dose 50, MESH: Bacterial Adhesion, MESH: Protein Kinases, MESH: Virulence Factors, MESH : Signal Transduction, MESH: Humans, Gene Expression Profiling, MESH: Transcription, Genetic, MESH : Gene Expression Profiling, MESH : Humans, MESH : Transcription, Genetic, Epithelial Cells, Gene Expression Regulation, Bacterial, MESH: Streptococcus agalactiae, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH : Promoter Regions (Genetics), MESH : Disease Models, Animal, Rats, Repressor Proteins, Disease Models, Animal, MESH: Lethal Dose 50, Genes, Bacterial, MESH: Gene Deletion, MESH : Animals, MESH: Disease Models, Animal, Protein Kinases, Gene Deletion
Abstract

International audience; In this study, we carried out a detailed structural and functional analysis of a Streptococcus agalactiae (GBS) two-component system which is orthologous to the CovS/CovR (CsrS/CsrR) regulatory system of Streptococcus pyogenes. In GBS, covR and covS are part of a seven gene operon transcribed from two promoters that are not regulated by CovR. A DeltacovSR mutant was found to display dramatic phenotypic changes such as increased haemolytic activity and reduced CAMP activity on blood agar. Adherence of the DeltacovSR mutant to epithelial cells was greatly increased and analysis by transmission electron microscopy revealed the presence at its surface of a fibrous extracellular matrix that might be involved in these intercellular interactions. However, the DeltacovSR mutant was unable to initiate growth in RPMI and its viability in human normal serum was greatly impaired. A major finding of this phenotypic analysis was that the CovS/CovR system is important for GBS virulence, as a 3 log increase of the LD(50) of the mutant strain was observed in the neonate rat sepsis model. The pleiotropic phenotype of the DeltacovSR mutant is in full agreement with the large number of genes controlled by CovS/CovR as seen by expression profiling analysis, many of which encode potentially secreted or cell surface-associated proteins: 76 genes are repressed whereas 63 were positively regulated. CovR was shown to bind directly to the regulatory regions of several of these genes and a consensus CovR recognition sequence was proposed using both DNase I footprinting and computational analyses.

Published
2004
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14. Interleukin-10 inhibits elevated chemokine interleukin-8 and regulated on activation normal T cell expressed and secreted production in cystic fibrosis bronchial epithelial cells by targeting the I(k)B kinase alpha/beta complex

Author

Céline Muselet, Jacky Jacquot, Dominique Hubert, Frank Antonicelli, Daniel Dusser, Sandie Escotte, Olivier Tabary, Dynamique cellulaire et moléculaire de la muqueuse respiratoire, Université de Reims Champagne-Ardenne (URCA)-IFR53-Institut National de la Santé et de la Recherche Médicale (INSERM), Croissance et réparation du poumon, Institut National de la Santé et de la Recherche Médicale (INSERM), Matrice extracellulaire et dynamique cellulaire - UMR 7369 (MEDyC), SFR CAP Santé (Champagne-Ardenne Picardie Santé), Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Université de Reims Champagne-Ardenne (URCA)-Centre National de la Recherche Scientifique (CNRS), Service de pneumologie [CHU Cochin], Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université de Reims Champagne-Ardenne (URCA) - IFR53 - Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS) - Université de Reims Champagne-Ardenne (URCA) - SFR CAP Santé (Champagne-Ardenne Picardie Santé), Université de Reims Champagne-Ardenne (URCA) - Université de Picardie Jules Verne (UPJV) - Université de Reims Champagne-Ardenne (URCA) - Université de Picardie Jules Verne (UPJV), Assistance publique - Hôpitaux de Paris (AP-HP) - CHU Cochin [AP-HP], Université de Reims Champagne-Ardenne (URCA)-SFR CAP Santé (Champagne-Ardenne Picardie Santé), Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS), and Birembaut, Philippe

Subjects
Chemokine, MESH: Interleukin-10, Cystic Fibrosis, MESH: I-kappa B Proteins, medicine.medical_treatment, Gene Expression, MESH: NF-kappa B, Sodium Chloride, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, Chemokine CCL5, Cells, Cultured, biology, NF-kappa B, Interleukin, MESH: Bronchi, MESH: Chemokines, MESH : Bronchi, I-kappa B Kinase, Interleukin-10, MESH : Epithelial Cells, Interleukin 10, Cytokine, medicine.anatomical_structure, MESH: Epithelial Cells, MESH : NF-kappa B, Tumor necrosis factor alpha, I-kappa B Proteins, Chemokines, MESH: Cells, Cultured, MESH : Protein-Serine-Threonine Kinases, medicine.medical_specialty, MESH : Interleukin-8, MESH: Gene Expression, MESH: Cystic Fibrosis, Macromolecular Substances, T cell, MESH : Macromolecular Substances, Bronchi, Respiratory Mucosa, Protein Serine-Threonine Kinases, MESH : Interleukin-1, MESH: Protein-Serine-Threonine Kinases, Pathology and Forensic Medicine, Internal medicine, MESH : Interleukin-10, MESH : Cystic Fibrosis, MESH : Cells, Cultured, medicine, Humans, MESH : Chemokines, Interleukin 8, MESH: I-kappa B Kinase, MESH: Humans, Tumor Necrosis Factor-alpha, Interleukin-8, MESH : Humans, MESH: Interleukin-1, Epithelial Cells, MESH : I-kappa B Kinase, MESH: Macromolecular Substances, Molecular biology, MESH: Interleukin-8, MESH : Gene Expression, Endocrinology, Cell culture, biology.protein, [SDV.MHEP.PSR] Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, MESH : I-kappa B Proteins, Interleukin-1, Regular Articles
Abstract

Free FULL TEXT http://ajp.amjpathol.org/cgi/reprint/162/1/293; Accumulating evidence suggests that in cystic fibrosis (CF) patients, airway fluids are characterized by decreased antibacterial activity, elevated NaCl concentration, and high levels of chemokines, resulting in exaggerated activation of the transcriptional nuclear factor (NF)-kappaB in airway epithelial cells. The present study was undertaken to evaluate the effects of anti-inflammatory cytokine interleukin-10 (IL-10) on NaCl-induced chemokine IL-8 and regulated on activation normal T cell expressed and secreted (RANTES) expression through the NF-kappaB signaling in primary deltaF508 CF and non-CF (control) human bronchial epithelial cells. Exposure of CF and non-CF bronchial epithelial cells to hypertonic (170 mmol/L NaCl) milieu compared to isotonic (115 mmol/L NaCl) and hypotonic (85 mmol/L NaCl) milieu caused a significant, NaCl-dependent increase in IL-8 and RANTES gene expression and protein production. Compared to non-CF cells, CF bronchial epithelial cells were characterized by a higher susceptibility to produce elevated IL-8 and RANTES production in an hypertonic NaCl milieu in response to IL-1beta activation. Treatment with IL-10 suppressed IL-8 and RANTES gene expression in both non-CF and CF bronchial epithelial cells was associated with a reduced expression of I(k)B (IKK) alpha/beta kinases, particularly for IKKalpha which is greater expressed in CF bronchial epithelial cells, and resulting in reduced NF-kappaB activation. These findings suggest that IL-10 might have anti-inflammatory benefits in airways of CF patients.

Published
2003

15. Early mitochondrial dysfunction, superoxide anion production, and DNA degradation are associated with non-apoptotic death of human airway epithelial cells induced by Pseudomonas aeruginosa exotoxin A

Author

Geraldo M. B. Pereira, Helvécio C. C. Póvoa, Edith Puchelle, Maria-Cristina Plotkowski, Gérard Lizard, Jean-Marie Tournier, Jean-Marie Zahm, Department of Microbiology and Immunology (DMI), UERJ-Laboratory of Electron Microscopy, Dynamique cellulaire et moléculaire de la muqueuse respiratoire, Université de Reims Champagne-Ardenne (URCA)-IFR53-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire des Lipoprotéines humaines et interactions vasculaires, Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratory of Immunopathology (LI), Laboratory of Immunopathology-UERJ, Department of Microbiology and Immunology ( DMI ), Université de Reims Champagne-Ardenne ( URCA ) -IFR53-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Laboratory of Immunopathology ( LI ), and Birembaut, Philippe

Subjects
MESH: Cell Death, MESH: ADP Ribose Transferases, MESH : DNA, Clinical Biochemistry, Cell, Apoptosis, MESH : Dose-Response Relationship, Drug, Mitochondrion, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, Membrane Potentials, MESH: Dose-Response Relationship, Drug, chemistry.chemical_compound, Superoxides, MESH: Intracellular Membrane, MESH : DNA Fragmentation, Respiratory system, Enzyme Inhibitors, Cells, Cultured, ADP Ribose Transferases, MESH : Cell Survival, Cell Death, Superoxide, MESH: DNA, MESH: Bronchi, Caspase Inhibitors, MESH : Bronchi, Mitochondria, MESH : Epithelial Cells, medicine.anatomical_structure, MESH: Cell Survival, MESH: Enzyme Inhibitors, MESH: Epithelial Cells, MESH : ADP Ribose Transferases, Intracellular, MESH: Cells, Cultured, Pulmonary and Respiratory Medicine, Programmed cell death, Cell Survival, Virulence Factors, Bacterial Toxins, Exotoxins, Bronchi, DNA Fragmentation, Respiratory Mucosa, Biology, Microbiology, Necrosis, Nasal Polyps, MESH : Cells, Cultured, medicine, Humans, MESH: DNA Fragmentation, MESH : Intracellular Membrane, Molecular Biology, MESH : Enzyme Inhibitors, MESH: Humans, MESH: Caspases, Dose-Response Relationship, Drug, MESH: Apoptosis, MESH : Humans, Epithelial Cells, Cell Biology, DNA, Intracellular Membranes, MESH: Exotoxins, chemistry, MESH: Bacterial Toxins, MESH : Exotoxins, MESH : Cell Death, MESH : Bacterial Toxins, Respiratory epithelium, [SDV.MHEP.PSR] Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, MESH : Caspases, MESH : Apoptosis, [ SDV.MHEP.PSR ] Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract
Abstract

It has been shown that bacterial exoproducts may induce airway epithelium injury. During the epithelial repair process, the respiratory epithelial cells no more establish tight junctional intercellular complexes and may be particularly susceptible to bacterial virulence factors. In this study, we analyzed the effect of Pseudomonas aeruginosa exotoxin A (ETA) at different periods of time and concentrations on 16 HBE 14o(-) human bronchial epithelial cells in culture conditions inducing a phenotype of repairing cells. ETA treatment for 24 and 48 h led to the killing of 40.0 +/- 5.7% and 79.0 +/- 1.4% of the cells, respectively, as determined by the dimethylthiazole 2,5 diphenyl tetrazolium bromide assay. At 1,000 ng/ml, ETA led to the killing of 25.2 +/- 6.6, 59.4 +/- 5.9, and 82.3 +/- 3.7% of the cells, after treatment periods of 7, 24, and 48 h, respectively. Cell death could not be inhibited by z-VAD-fmk, a broad spectrum caspase inhibitor. By transmission electron microscopy, ultrastructural characteristics described in apoptosis were not detected in ETA-treated cells. Instead, the mitochondria of cells treated for 24 and 48 h with ETA at 100 and 1,000 ng/ml were highly condensed. Human nasal polyp epithelial cells in primary culture exposed to ETA at 1,000 ng/ml did not exhibit characteristic features of apoptotic cells either. Cytofluorometric analysis of ETA-treated 16 HBE 14o(-) cells labeled with DiOC(6)(3) and hydroethidine showed a time- and dose-dependent reduction of the mitochondrial transmembrane potential, detected 7 h after ETA treatment, and an increase in superoxide production, detected at 24 h, respectively. By a photometric assay, DNA degradation was also detected 7 h after cell treatment with ETA at 100 and 1,000 ng/ml. Taken together, our results show that ETA-induced death of epithelial respiratory cells was preceded by early mitochondrial dysfunction and superoxide anion production, but was not followed by the classically described apoptotic pathways.

Published
2002

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