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24 results on '"MESH : DNA Primers"'

1. BAT3 modulates p300-dependent acetylation of p53 and autophagy-related protein 7 (ATG7) during autophagy

Author

Laetitia K. Linares, Andrew V. Hubberstey, Emmanuelle Liaudet-Coopman, Nelly Pirot, Fabienne Desmots, Guy Berchem, Anne-Sophie Bach, Esther Pérez-Gracia, Christine Prébois, Valérie Palissot, Patrice Codogno, Céline Gongora, Salwa Sebti, Sophie Pattingre, Chantal Bauvy, 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 ), Institut Necker Enfants-Malades (INEM) ( INEM - UM 111 (UMR 8253 / U1151) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Service d'Hématologie, Immunologie et de Thérapie Cellulaire ( HITC ), Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -Hôpital Pontchaillou-CHU Pontchaillou [Rennes], Department of Biological Sciences [Windsor], University of Windsor [Ca], Laboratory of Experimental Hemato-Oncology ( CRP-Santé ), Centre de Recherche Public-Santé, This work is supported by INSERM, ARC (SP), La Ligue Régionale contre Le Cancer (Comités du Gard et de l'Hérault) (SP), Cancéropole GSO (SP) and CHEMORES consortium (EU FP6, ELC). SS has fellowships from the FNR Luxembourg and La Ligue Nationale contre le Cancer., Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), 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), Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'Hématologie, Immunologie et de Thérapie Cellulaire (HITC), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Hôpital Pontchaillou-CHU Pontchaillou [Rennes], Laboratory of Experimental Hemato-Oncology (CRP-Santé), Le Ster, Yves, 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), and Université de Rennes (UR)-Hôpital Pontchaillou-CHU Pontchaillou [Rennes]

Subjects
p53, Autophagy-Related Protein 7, MESH: Mice, Knockout, Mice, Cytosol, MESH: Cytosol, MESH : Embryo, Mammalian, MESH: Animals, Nuclear protein, MESH: Tumor Suppressor Protein p53, Mice, Knockout, MESH : Cell Nucleus, Multidisciplinary, MESH: Real-Time Polymerase Chain Reaction, MESH : Cytosol, Life Sciences, Nuclear Proteins, Biological Sciences, Cell biology, medicine.anatomical_structure, MESH : Cell Fractionation, MESH: Cell Fractionation, MESH : DNA Primers, MESH: Molecular Chaperones, Microtubule-Associated Proteins, MESH: Acetylation, MESH: Cell Nucleus, MESH: DNA Primers, MESH : Real-Time Polymerase Chain Reaction, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, Cell Fractionation, Real-Time Polymerase Chain Reaction, MESH : Acetylation, BAG3, MESH : E1A-Associated p300 Protein, MESH : Immunoprecipitation, BAT3, MESH : Mice, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Autophagy, medicine, Animals, Immunoprecipitation, MESH: Autophagy, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, MESH: Mice, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, DNA Primers, acetylation, Cell Nucleus, MESH: Immunoprecipitation, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, MESH: Embryo, Mammalian, MESH : Molecular Chaperones, MESH : Nuclear Proteins, Embryo, Mammalian, MESH : Autophagy, Molecular biology, ATG, MESH: Microtubule-Associated Proteins, MESH : Tumor Suppressor Protein p53, Cell nucleus, MESH : Microtubule-Associated Proteins, Acetylation, MESH : Mice, Knockout, MESH: E1A-Associated p300 Protein, MESH : Animals, Tumor Suppressor Protein p53, E1A-Associated p300 Protein, MESH: Nuclear Proteins, Nuclear localization sequence, Molecular Chaperones
Abstract

International audience; Autophagy is regulated by posttranslational modifications, including acetylation. Here we show that HLA-B-associated transcript 3 (BAT3) is essential for basal and starvation-induced autophagy in embryonic day 18.5 BAT3(-/-) mouse embryos and in mouse embryonic fibroblasts (MEFs) through the modulation of p300-dependent acetylation of p53 and ATG7. Specifically, BAT3 increases p53 acetylation and proautophagic p53 target gene expression, while limiting p300-dependent acetylation of ATG7, a mechanism known to inhibit autophagy. In the absence of BAT3 or when BAT3 is located exclusively in the cytosol, autophagy is abrogated, ATG7 is hyperacetylated, p53 acetylation is abolished, and p300 accumulates in the cytosol, indicating that BAT3 regulates the nuclear localization of p300. In addition, the interaction between BAT3 and p300 is stronger in the cytosol than in the nucleus and, during starvation, the level of p300 decreases in the cytosol but increases in the nucleus only in the presence of BAT3. We conclude that BAT3 tightly controls autophagy by modulating p300 intracellular localization, affecting the accessibility of p300 to its substrates, p53 and ATG7.

Published
2014
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2. Aberrant DNA methylation distinguishes hepatocellular carcinoma associated with HBV and HCV infection and alcohol intake

Author

Fabien Zoulim, Marie-Pierre Lambert, Massimo Tommasino, Zdenko Herceg, Jörg Tost, Jean-Yves Scoazec, Anupam Paliwal, Pierre Hainaut, Thomas Vaissière, Bakary S. Sylla, Isabelle Chemin, Centre de Recherche en Cancérologie de Lyon (CRCL), Centre Léon Bérard [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), Interactions Bactéries-Cellules (UIBC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris]-Institut National de la Recherche Agronomique (INRA), International Agency for Cancer Research (IACR), Equipe 16, Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Service d'hépatologie et de gastroentérologie, Hospices Civils de Lyon (HCL)-Hôtel-Dieu-Hospices Civils de Lyon (HCL)-Hôtel-Dieu-Centre de Recherche en Cancérologie de Lyon (CRCL), Centre International de Recherche contre le Cancer - International Agency for Research on Cancer (CIRC - IARC), Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Sect Mech Carcinogenesis, equipe 4, Laboratoire Central d'Anatomie et de Cytologie Pathologiques [Hôpital Edouard Herriot - HCL], Hôpital Edouard Herriot [CHU - HCL], Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL)-Hôpital Edouard Herriot [CHU - HCL], Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL)-Centre de Recherche en Cancérologie de Lyon (CRCL), Centre National de Génotypage (CNG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Centre de Recherche en Cancérologie de Lyon ( CRCL ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Interactions Bactéries-Cellules ( UIBC ), Institut National de la Recherche Agronomique ( INRA ) -Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), International Agency for Cancer Research ( IACR ), International Agency for Cancer Research, Université de Lyon-Université de Lyon-Centre Léon Bérard [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 ), Hospices Civils de Lyon ( HCL ) -Hôtel-Dieu-Hospices Civils de Lyon ( HCL ) -Hôtel-Dieu-Centre de Recherche en Cancérologie de Lyon ( CRCL ), Centre International de Recherche contre le Cancer - International Agency for Research on Cancer ( CIRC - IARC ), Organisation mondiale de la santé (OMS/WHO), international Agency for Research on Cancer - IARC, Hospices Civils de Lyon ( HCL ) -Hospices Civils de Lyon ( HCL ) -Hôpital Edouard Herriot [CHU - HCL], Hospices Civils de Lyon ( HCL ) -Hospices Civils de Lyon ( HCL ) -Centre de Recherche en Cancérologie de Lyon ( CRCL ), Centre National de Génotypage ( CNG ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Hospices Civils de Lyon (HCL)-Hôtel-Dieu-Hospices Civils de Lyon (HCL)-Hôtel-Dieu-Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Hospices Civils de Lyon (HCL)-Hospices Civils de Lyon (HCL)-Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), and Institut National de la Recherche Agronomique (INRA)-Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Male, MESH : Liver Neoplasms, MESH : Aged, Receptors, Nicotinic, MESH: Base Sequence, medicine.disease_cause, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, MESH: DNA Methylation, 0302 clinical medicine, Risk Factors, MESH: Liver Neoplasms, MESH: Risk Factors, MESH : Female, MESH: Gene Silencing, MESH: Carcinoma, Hepatocellular, MESH: Glutathione S-Transferase pi, MESH: Aged, 0303 health sciences, MESH: Middle Aged, MESH : Tumor Suppressor Proteins, Liver Neoplasms, RNA-Binding Proteins, DNA, Neoplasm, Hepatitis C, Methylation, Middle Aged, MESH : Adult, Hepatitis B, MESH : Risk Factors, 3. Good health, DNA-Binding Proteins, 030220 oncology & carcinogenesis, Hepatocellular carcinoma, DNA methylation, MESH: Receptors, Nicotinic, Female, MESH : DNA Primers, MESH : DNA, Neoplasm, MESH : DNA-Binding Proteins, Adult, MESH : Carcinoma, Hepatocellular, MESH: DNA Primers, Carcinoma, Hepatocellular, Alcohol Drinking, MESH : Male, Hepatitis C virus, MESH : RNA-Binding Proteins, MESH: DNA, Neoplasm, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, MESH: Phosphoproteins, 03 medical and health sciences, MESH : Gene Silencing, medicine, Humans, MESH : Middle Aged, MESH: Tumor Suppressor Proteins, Gene Silencing, neoplasms, Aged, DNA Primers, 030304 developmental biology, MESH: Hepatitis C, Hepatitis B virus, MESH: Humans, Base Sequence, MESH : Receptors, Nicotinic, MESH: Hepatitis B, Hepatology, Tumor Suppressor Proteins, MESH : Humans, MESH : Hepatitis B, Cancer, MESH: Adult, DNA Methylation, MESH : Hepatitis C, Phosphoproteins, medicine.disease, MESH: Male, digestive system diseases, MESH : Alcohol Drinking, MESH: RNA-Binding Proteins, Glutathione S-Transferase pi, MESH : Glutathione S-Transferase pi, Immunology, MESH : Base Sequence, MESH : Phosphoproteins, MESH : DNA Methylation, MESH: Female, MESH: Alcohol Drinking, MESH: DNA-Binding Proteins
Abstract

International audience; BACKGROUND & AIMS: Hepatocellular carcinoma (HCC) is one of the most frequent human cancers and a major cause of cancer-related death worldwide. The major risk factors for developing HCC are infection by hepatitis B virus (HBV) and hepatitis C virus (HCV), chronic alcoholism, and aflatoxins; however, critical gene targets remain largely unknown. Herein, we sought to establish DNA methylation patterns in HCC and corresponding cirrhotic tissues and to identify DNA methylation changes associated with major risk factors. METHODS: We have established assays for quantitative analysis of DNA methylation levels in a panel of seven cancer-associated genes and repetitive elements, and combined these assays with a series of HCC tumors, associated with major risk factors, collected from two different geographical areas. RESULTS: We found a high frequency of aberrant hypermethylation of specific genes (RASSF1A, GSTP1, CHRNA3, and DOK1) in HCC tumors as compared to control cirrhotic or normal liver tissues, suggesting that aberrant hypermethylation exhibits non-random and tumor-specific patterns in HCC. Importantly, our analysis revealed an association between alcohol intake and the hypomethylation of MGMT and between hypermethylation of GSTP1 and HBV infection, indicating that hypermethylation of the genes analyzed in HCC tumors exhibits remarkably distinct patterns depending on associated risk factors. CONCLUSIONS: This study identifies aberrant DNA methylation of specific cellular genes in HCC and the major risk factors associated with these changes, providing information that could be exploited for biomarker discovery in clinics and molecular epidemiology.

Published
2011
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3. Proteomic analysis and immunogenicity of secreted proteins from Rhodococcus equi ATCC 33701

Author

Aurélie Budin-Verneuil, Axel Hartke, Vianney Pichereau, Séverine Cauchard, Claire Laugier, Corinne Barbey, Sandrine Petry, Unité de Recherche Risques Microbiens ( U2RM ), Université de Caen Normandie ( UNICAEN ), Normandie Université ( NU ) -Normandie Université ( NU ), Laboratoire de Pathologie équine de Dozulé, ANSES, Laboratoire des Sciences de l'Environnement Marin (LEMAR) ( LEMAR ), Centre National de la Recherche Scientifique ( CNRS ) -Université de Brest ( UBO ) -Institut Français de Recherche pour l'Exploitation de la Mer ( IFREMER ) -Institut Universitaire Européen de la Mer ( IUEM ), Institut de Recherche pour le Développement ( IRD ) -Université de Brest ( UBO ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université de Brest ( UBO ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ), Unité de Recherche Risques Microbiens (U2RM), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Laboratoire d'études et de recherches en pathologie équine, Agence Française de Sécurité Sanitaire des Aliments (AFSSA), Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), and Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Mycobacterium tuberculosis, Proteome, MESH: Sequence Homology, Amino Acid, MESH: Trypsin, MESH : Enzymes, MESH: Amino Acid Sequence, Proteomics, Mass Spectrometry, [ SDV.MP ] Life Sciences [q-bio]/Microbiology and Parasitology, MESH : Actinomycetales Infections, MESH : Bacterial Proteins, Trypsin, MESH: Animals, Rhodococcus equi, MESH: Bacterial Proteins, MESH : Temperature, Peptide sequence, MESH : Immunoblotting, 0303 health sciences, MESH: Immunoblotting, MESH : Horse Diseases, MESH : Amino Acid Sequence, Immunogenicity, MESH : Antigens, Bacterial, Temperature, General Medicine, MESH: Temperature, MESH : Sequence Homology, Amino Acid, Enzymes, 3. Good health, MESH: Proteome, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Electrophoresis, Polyacrylamide Gel, MESH : DNA Primers, Actinomycetales Infections, MESH: DNA Primers, Signal peptide, MESH: Rhodococcus equi, Sequence analysis, Immunoblotting, MESH: Enzymes, MESH : Proteome, Biology, MESH : Electrophoresis, Polyacrylamide Gel, Microbiology, MESH : Mycobacterium tuberculosis, 03 medical and health sciences, MESH : Mass Spectrometry, Bacterial Proteins, Animals, Amino Acid Sequence, Horses, MESH: Horses, DNA Primers, 030304 developmental biology, MESH: Mass Spectrometry, Antigens, Bacterial, Sequence Homology, Amino Acid, General Veterinary, MESH : Rhodococcus equi, 030306 microbiology, Mycobacterium tuberculosis, biology.organism_classification, Secretory protein, MESH : Horses, Horse Diseases, MESH : Trypsin, MESH : Animals, MESH: Horse Diseases, MESH: Antigens, Bacterial, MESH: Actinomycetales Infections, MESH: Electrophoresis, Polyacrylamide Gel
Abstract

International audience; Rhodococcus equi is one of the most important causes of mortality in foals between 1 and 6 months of age. Although rare, infection also occurs in a variety of other mammals including humans, often following immunosuppression of various causes. Secreted proteins are known to mediate important pathogen-host interactions and consequently are favored candidates for vaccine development as they are the most easily accessible microbial antigens to the immune system. Here, we describe the results of a proteomic analysis based on SDS-PAGE, immunoblot and mass spectrometry, which was carried out aiming the identification of secreted proteins that are differently expressed at 30 degrees C versus 37 degrees C and at mid-exponential versus early-stationary growth phase and antigenic proteins from R. equi ATCC 33701. A total of 48 proteins was identified regardless of growth conditions. The cholesterol oxidase ChoE appears to be the major secretory protein. Moreover, four proteins revealed high homologies with the mycolyl transferases of the Ag85 complex from Mycobacterium tuberculosis. The sequence analysis predicted that 24 proteins are transported by a signal peptide-dependent pathway. Moreover, five antigenic proteins of R. equi were identified by immunoblot, including a novel strongly immunoreactive protein of unknown function. In conclusion, the elucidation of the secretome of R. equi identified several proteins with different biological functions and a new candidate for developing vaccines against R. equi infection in horse.

Published
2009
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4. One-Step Multiplex PCR Assay for Differentiating Proposed New Species 'Clostridium neonatale' from Closely Related Species

Author

Philippe Bouvet, Marie Jose Butel, Sophia Schönherr, Laurent Ferraris, Julio Aires, Brunhilde Dauphin, Michel R. Popoff, Ecosystème intestinal, probiotiques, antibiotiques (EA 4065), Université Paris Descartes - Paris 5 (UPD5), Bactéries anaérobies et Toxines, Institut Pasteur [Paris], Andromas SAS, Institut Pasteur [Paris] (IP), Ecosystème intestinal, probiotiques, antibiotiques ( EA 4065 ), and Université Paris Descartes - Paris 5 ( UPD5 )

Subjects
[SDV]Life Sciences [q-bio], [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, law.invention, Disease Outbreaks, Feces, Clostridium, law, RNA, Ribosomal, 16S, MESH : Clostridium, MESH : Multiplex Polymerase Chain Reaction, MESH : DNA, Bacterial, Multiplex, MESH: Disease Outbreaks, Polymerase chain reaction, MESH : Intestines, Cross Infection, biology, MESH: Multiplex Polymerase Chain Reaction, MESH: Infant, Newborn, MESH: Feces, 3. Good health, Intestines, MESH: RNA, Ribosomal, 16S, MESH : Canada, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Necrotizing enterocolitis, MESH : DNA Primers, Variants of PCR, MESH : Cross Infection, MESH: Intestines, Microbiology (medical), DNA, Bacterial, MESH: DNA Primers, Canada, Clostridium neonatale, MESH: Enterocolitis, Necrotizing, MESH : Intensive Care Units, Neonatal, MESH : Infant, Newborn, Microbiology, Enterocolitis, Necrotizing, MESH: Canada, Intensive Care Units, Neonatal, Multiplex polymerase chain reaction, parasitic diseases, medicine, Humans, MESH : Disease Outbreaks, DNA Primers, MESH: Humans, [ SDV ] Life Sciences [q-bio], MESH: Clostridium, MESH : Humans, Infant, Newborn, Outbreak, MESH: Cross Infection, Bacteriology, MESH : Feces, biology.organism_classification, medicine.disease, bacterial infections and mycoses, MESH: DNA, Bacterial, MESH : RNA, Ribosomal, 16S, MESH : Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, MESH: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Multiplex Polymerase Chain Reaction, MESH : Enterocolitis, Necrotizing, MESH: Intensive Care Units, Neonatal
Abstract

“ Clostridium neonatale ” sp. nov., previously involved in an outbreak of neonatal necrotizing enterocolitis, was recently proposed as a new species of the Clostridium genus sensu stricto . We developed a one-step multiplex colony PCR for C. neonatale identification and investigated C. neonatale intestinal colonization frequency in healthy preterm neonates.

Published
2015
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5. Delta-like 4 inhibits choroidal neovascularization despite opposing effects on vascular endothelium and macrophages. : DLL4's opposing effects in choroidal neovascularization

Author

Camelo, Serge, Raoul, William, Lavalette, Sophie, Calippe, Bertrand, Cristofaro, Brunella, Levy, Olivier, Houssier, Marianne, Sulpice, Eric, Jonet, Laurent, Klein, Christophe, Devevre, Estelle, Thuret, Gilles, Duarte, Antonio, Eichmann, Anne, Leconte, Laurence, Guillonneau, Xavier, Sennlaub, Florian, Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -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 de la Vision, 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 ), Centre interdisciplinaire de recherche en biologie ( CIRB ), Collège de France ( CdF ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Biologie à Grande Échelle ( BGE - UMR S1038 ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Grenoble Alpes [Saint Martin d'Hères]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Faculté de Médecine, CHU Saint-Etienne-Université Jean Monnet [Saint-Étienne] ( UJM ) -Assistance Publique des Hôpitaux de St Etienne, Instituto Gulbenkian de Ciência [Oeiras] ( IGC ), Fundação Calouste Gulbenkian, Faculdade de Medicina Veterinária ( CIISA ), Universidade Técnica de Lisboa, Centre de Recherche Cardiovasculaire de Lariboisiere, Université Paris Diderot - Paris 7 ( UPD7 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Département d'Ophtalmologie, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôtel-Dieu, ANR blanc AO5120DD ANR Genopat R09099DS ERC-2007 Starting Grant 210345, Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), 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), Centre interdisciplinaire de recherche en biologie (CIRB), Labex MemoLife, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie à Grande Échelle (BGE - UMR S1038), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), CHU Saint-Etienne-Université Jean Monnet [Saint-Étienne] (UJM)-Assistance Publique des Hôpitaux de St Etienne, Instituto Gulbenkian de Ciência [Oeiras] (IGC), Faculdade de Medicina Veterinária (CIISA), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôtel-Dieu, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Faculté de Médecine Jacques Lisfranc [Université Saint-Etienne], and Université Jean Monnet - Saint-Étienne (UJM)

Subjects
MESH: DNA Primers, Notch, genetic structures, age related macular degeneration, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH : Blotting, Western, MESH : Mice, Inbred C57BL, MESH: Base Sequence, DLL4, MESH: Mice, Inbred C57BL, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Mice, MESH: Blotting, Western, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [SDV.MHEP.OS]Life Sciences [q-bio]/Human health and pathology/Sensory Organs, MESH: Intercellular Signaling Peptides and Proteins, MESH: Mice, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Intercellular Signaling Peptides and Proteins, MESH : Choroidal Neovascularization, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, MESH: Choroidal Neovascularization, MESH : Reverse Transcriptase Polymerase Chain Reaction, eye, eye diseases, macrophages, MESH : Endothelium, Vascular, MESH : Macrophages, Peritoneal, [ SDV.MHEP.OS ] Life Sciences [q-bio]/Human health and pathology/Sensory Organs, MESH : Base Sequence, MESH : DNA Primers, MESH: Endothelium, Vascular, sense organs, Angiogenesis, MESH : Animals, MESH: Macrophages, Peritoneal
Abstract

International audience; Inflammatory neovascularization, such as choroidal neovascularization (CNV), occur in the presence of Notch expressing macrophages. DLL4s anti-angiogenic effect on endothelial cells (EC) has been widely recognized, but its influence on Notch signaling on macrophages and its overall effect in inflammatory neovascularization is not well understood. We identified macrophages and ECs as the main Notch 1 and Notch 4 expressing cells in CNV. A soluble fraction spanning Ser28-Pro525 of the murine extracellular DLL4 domain (sDLL4/28-525) activated the Notch pathway, as it induces Notch target genes in macrophages and ECs and inhibited EC proliferation and vascular sprouting in aortic rings. In contrast, sDLL4/28-525 increased pro-angiogenic VEGF, and IL-1β expression in macrophages responsible for increased vascular sprouting observed in aortic rings incubated in conditioned media from sDLL4/28-525 stimulated macrophages. In vivo, Dll4(+/-) mice developed significantly more CNV and sDLL4/28-525 injections inhibited CNV in Dll4(+/-) CD1 mice. Similarly, sDLL4/28-525 inhibited CNV in C57Bl6 and its effect was reversed by a γ-secretase inhibitor that blocks Notch signaling. The inhibition occurred despite increased VEGF, IL-1β expression in infiltrating inflammatory macrophages in sDLL4/28-525 treated mice and might be due to direct inhibition of EC proliferation in laser-induced CNV as demonstrated by EdU labelling in vivo. In conclusion, Notch activation on macrophages and ECs leads to opposing effects in inflammatory neovascularization in situations such as CNV.

Published
2012
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6. Natural polymorphisms of HIV-1 CRF01_AE integrase coding region in ARV-naive individuals in Cambodia, Thailand and Vietnam : an ANRS AC12 working group study

Author

Martine Peeters, Eric Nerrienet, Tawee Donchai, Ahidjo Ayouba, Ton Tran, Janin Nouhin, Marie-Laure Chaix, Nicole Ngo-Giang-Huong, Sreymom Ken, Truong Xuan Lien, Khanh Thu Huynh Hoang, Jiraporn Kamkorn, Institut Pasteur du Cambodge, Réseau International des Instituts Pasteur (RIIP), Épidémiologie clinique, santé mère-enfant et VIH en Asie du Sud-Est (IRD_PHPT), Harvard University-Chiang Mai University (CMU), Institut Pasteur d'Ho Chi Minh Ville, VIH/SIDA et maladies associées, Université Montpellier 1 (UM1), Infections à Vih, Réservoirs, Pharmacologie des Antirétroviraux et Prévention de la Transmission Mère Enfant, Université Paris Descartes - Paris 5 (UPD5), Immunology and Infectious Diseases, Harvard School of Public Health, Harvard University [Cambridge]-Chiang Mai university (Thaïlande), Institut Pasteur du Cambodge-Réseau International des Instituts Pasteur ( RIIP ), Épidémiologie clinique, santé mère-enfant et VIH en Asie du Sud-Est ( IRD_PHPT ), Réseau International des Instituts Pasteur ( RIIP ) -Institut Pasteur d'Ho Chi Minh Ville, Université Montpellier 1 ( UM1 ), Université Paris Descartes - Paris 5 ( UPD5 ), and Harvard University [Cambridge]-Chiang Mai University (CMU)

Subjects
MESH: CD4 Lymphocyte Count, MESH : Polymorphism, Genetic, Integrase inhibitor, HIV Infections, HIV Integrase, Drug resistance, MESH: Base Sequence, Polymerase Chain Reaction, MESH: HIV-1, 0302 clinical medicine, 030212 general & internal medicine, MESH: Anti-HIV Agents, MESH : Polymerase Chain Reaction, MESH : Algorithms, Genetics, 0303 health sciences, MESH: HIV Infections, Thailand, MESH: Amino Acid Substitution, 3. Good health, Integrase, Infectious Diseases, Vietnam, Lentivirus, MESH : DNA Primers, Cambodia, Algorithms, MESH : HIV-1, medicine.drug, MESH: DNA Primers, Microbiology (medical), Anti-HIV Agents, MESH: Algorithms, Biology, South East Asia, Microbiology, Virus, 03 medical and health sciences, MESH : Amino Acid Substitution, Resistance mutations, [ SDV.MHEP ] Life Sciences [q-bio]/Human health and pathology, HIV-1 integrase, MESH: Polymorphism, Genetic, MESH : CD4 Lymphocyte Count, MESH : HIV Infections, Consensus sequence, medicine, Humans, MESH: Thailand, Molecular Biology, Ecology, Evolution, Behavior and Systematics, DNA Primers, 030304 developmental biology, Polymorphism, Genetic, MESH: Humans, Base Sequence, MESH : Cambodia, MESH : Anti-HIV Agents, MESH: Cambodia, MESH : Humans, MESH: Polymerase Chain Reaction, MESH : Vietnam, MESH : Thailand, biology.organism_classification, Raltegravir, Virology, CD4 Lymphocyte Count, Multiple drug resistance, Amino Acid Substitution, HIV-1, biology.protein, MESH : Base Sequence, MESH: Vietnam, MESH: HIV Integrase, Antiretroviral naive, MESH : HIV Integrase, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

International audience; The HIV integrase enzyme is essential for the HIV life cycle as it mediates integration of HIV-1 proviral DNA into the infected cell's genome. Recently, the development of drugs capable of inhibiting integrase has provided major new options for HIV-infected, treatment-experienced patients with multidrug resistant virus, as well treatment-naïve patients. More than 40 amino acid substitutions within integrase have been described as associated mostly with resistance of HIV B-subtypes to currently available integrase inhibitors (INIs). We have analyzed the natural polymorphisms of the integrase coding region in 87 antiretroviral-naïve subjects (32 from Cambodia, 37 from Thailand and 18 from Vietnam) infected with CRF01_AE virus, the predominant HIV-1 strain circulating in Southeast Asia. The 864bp integrase coding region was sequenced using the ANRS consensus sequencing technique from plasma samples, and amino acid results were interpreted for drug resistance according to the ANRS (Updated July 2009, version 18) and Stanford algorithms (Version November 6, 2009). Alignment of the 87 amino acid sequences against the 2004 Los Alamos HIV-1 clade B consensus sequence showed that overall, 119 of 288 (41.3%) amino acid positions presented at least one polymorphism each. Substitutions found in >60% of study subjects occurred at: K14, A21, V31, S39, I72, T112, T124, T125, G134, I135, K136, D167, V201, L234 and S283. Also, new amino acid substitutions of as yet unknown significance were identified: E152K/H, S153F/L, N155I and E157G. None of the known integrase resistance mutations were observed, except E157Q found in one Cambodian subject (1.1%, CI 95% 0.02-6.3%). The clinical impact of this substitution on resistance of B and nonB-viruses to the licensed INI raltegravir is unclear. If this substitution is confirmed to compromise the virologic response to raltegravir, further studies will be needed to better assess the prevalence of this substitution among CRF01_AE virus.

Published
2011
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7. Placental and fetal steroidogenesis

Author

Sanderson , J Thomas, Institut Armand Frappier ( INRS-IAF ), and Institut National de la Recherche Scientifique [Québec] ( INRS ) -Réseau International des Instituts Pasteur ( RIIP ) -Institut Armand Frappier

Subjects
MESH : Pregnancy, MESH : RNA, Messenger, MESH : Tumor Cells, Cultured, MESH : Placenta, MESH : Humans, [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, MESH : Aromatase, MESH : Reverse Transcriptase Polymerase Chain Reaction, MESH : Base Sequence, MESH : DNA Primers, MESH : Female, MESH : Fetus, MESH : Steroids
Abstract

International audience; Steroid hormones are essential for maintenance of pregnancy and fetal development. The expression and catalytic activity of the key steroidogenic enzymes involved in the production of progesterone and estrogens increase during pregnancy, and there is an intricate communication between the mother, the placenta, and the fetus in order to maintain a balanced supply of the steroid hormones essential for embryogenesis. This chapter describes methods for the measurement of the expression and catalytic activity of three key cytochrome P450 (CYP) enzymes involved in the production of progesterone and estrogens, aromatase (CYP19), steroid 17-hydroxylase/17,20-lyase (CYP17), and cholesterol side-chain cleavage (CYP11A).

Published
2009
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8. In vivo and ex vivo gene transfer in thymocytes and thymocyte precursors

Author

Adjali , Oumeya, Montel-Hagen , Amélie, Swainson , Louise, Marty , Sophie, Vicente , Rita, Mongellaz , Cedric, Jacquet , Chantal, Zimmermann , Valérie, Taylor , Naomi, Institut de Génétique Moléculaire de Montpellier (IGMM), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), Institut de Génétique Moléculaire de Montpellier ( IGMM ), and Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
MESH : Cell Line, MESH: DNA Primers, MESH: Gene Transfer Techniques, Antigens, CD34, MESH : Transduction, Genetic, Thymus Gland, MESH: Base Sequence, Cell Line, Mice, Transduction, Genetic, MESH : Mice, Animals, Humans, MESH : Thymus Gland, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Gene Transfer Techniques, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Mice, DNA Primers, MESH: Humans, Base Sequence, MESH : Humans, Gene Transfer Techniques, MESH: Antigens, CD34, MESH: Thymus Gland, MESH: Transduction, Genetic, MESH: Cell Line, MESH : Antigens, CD34, MESH : Base Sequence, MESH : DNA Primers, MESH : Animals
Abstract

International audience; The thymus provides a specialized environment allowing the differentiation of T lymphocytes from bone marrow-derived progenitor cells. We and others have demonstrated that gene transfer into distinct thymocyte populations can be obtained, both in vivo and ex vivo, using lentiviral vectors. Here, we describe techniques for intrathymic lentiviral transduction in mice, using a surgical approach wherein the thoracic cavity is exposed as well as a significantly less invasive strategy wherein virions are directly injected through the skin. Moreover, thymocyte differentiation from murine and human progenitors is now feasible in vitro, under conditions wherein the Notch and IL-7 signaling pathways are activated. We describe methods allowing transduction of murine and human progenitors and their subsequent differentiation into more mature thymocytes. Conditions for lentiviral gene transfer into more differentiated human thymocyte subsets are also presented. Optimization of technologies for HIV-based gene transfer into murine and human thymocyte progenitors will advance strategies aimed at modulating T-cell differentiation and function in-vivo; approaches potentially targeting patients with genetic and acquired immunodeficiencies as well as immune-sensitive tumors. Furthermore, this technology will foster the progression of basic research aimed at elucidating molecular aspects of T-cell differentiation in mice and humans.

Published
2008
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9. [Role of the Regulatory T lymphocytes in hepatitis C fibrosis progression]

Author

Delhem, Nadira, Cottrez, Françoise, Carpentier, Arnaud, Miroux, Céline, Moralès, Olivier, François, Violaine, Groux, Hervé, Auriault, Claude, Pancré, Véronique, Institut de Biologie de Lille, Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Centre National de la Recherche Scientifique (CNRS), Immunosearch, [Institut Cochin] Departement Infection, immunité, inflammation, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-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 de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 2019) (UNS), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS), Réseau International des Instituts Pasteur ( RIIP ) -Réseau International des Instituts Pasteur ( RIIP ) -Centre National de la Recherche Scientifique ( CNRS ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -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 de pharmacologie moléculaire et cellulaire ( IPMC ), Université Nice Sophia Antipolis ( UNS ), and Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
Liver Cirrhosis, MESH : Liver Neoplasms, MESH: Interleukin-10, MESH : Cytokines, Biopsy, [ SDV.TOX ] Life Sciences [q-bio]/Toxicology, MESH : Aged, MESH : T-Lymphocytes, Helper-Inducer, Hepacivirus, T-Lymphocytes, Regulatory, MESH : Hepacivirus, MESH : T-Lymphocytes, Regulatory, MESH: Biopsy, Transforming Growth Factor beta, MESH: Liver Neoplasms, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH: Hepacivirus, [ SDV.IB ] Life Sciences [q-bio]/Bioengineering, MESH: Carcinoma, Hepatocellular, MESH: Antigens, CD, MESH: Aged, MESH: Cytokines, MESH: Middle Aged, MESH : Immune Tolerance, Liver Neoplasms, T-Lymphocytes, Helper-Inducer, MESH : Adult, Middle Aged, Hepatitis C, Interleukin-10, MESH: Hepatitis C, Chronic, Liver, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Disease Progression, Cytokines, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH : DNA Primers, MESH: Disease Progression, [SDV.IB]Life Sciences [q-bio]/Bioengineering, MESH: Liver Cirrhosis, MESH : Carcinoma, Hepatocellular, Adult, MESH: DNA Primers, Carcinoma, Hepatocellular, MESH: Immune Tolerance, MESH : Hepatitis C, Chronic, Antigens, CD, MESH : Interleukin-10, MESH : Antigens, CD, Immune Tolerance, MESH : Liver Cirrhosis, Humans, MESH : Middle Aged, MESH: T-Lymphocytes, Helper-Inducer, MESH: Transforming Growth Factor beta, Aged, DNA Primers, MESH: Hepatitis C, MESH: Humans, MESH: T-Lymphocytes, Regulatory, MESH : Humans, MESH : Liver, MESH: Adult, MESH : Disease Progression, MESH : Hepatitis C, Hepatitis C, Chronic, MESH : Transforming Growth Factor beta, MESH : Biopsy, MESH: Liver
Abstract

International audience; Hepatitis C virus (HCV) becomes chronic in about 85 % of infected individuals, whereas only 15 % of infected people clear spontaneously the virus. The progression of hepatitis C to chronic status is associated to a profound down-regulation of CD4 and CD8 multispecific immune response. This immune defect may participate to the immune tolerance of VHC and consequently to its persistence. Recent findings indicate that T regulatory cells as Tr1 play an inhibitory role on T helper responses notably in the context of auto-immune or inflammatory disorders. The existence of immunosuppressive mechanisms supported by Tr1 lymphocytes and their IL-10 production represent an attractive hypothesis. We have previously evaluated the existence of regulatory T cells (Tr1) via high production of IL-10, in liver biopsies of three well-defined cohorts of HCV-1b infected patients. To this purpose, we compared liver biopsies of chronically infected patients including patients without liver lesions, with cirrhosis and with hepatocellular carcinoma (HCC). Using quantitative real time PCR, the results obtained demonstrate, an increased expression of interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta)_, in liver biopsies with more severe fibrosis. This observation was correlated with an increased expression during the pathogenesis progression, of the three specific markers of the Tr1 cells sub-population, recently described and confirming the Tr1 phenotype. Evidence of regulatory T cells installation in the liver of chronically infected patient and increased frequency in cirrhosis and HCC suggest a main role of these cells in the aggravation of the liver pathology. This study should bring insight of T regulatory cell implications in VHC persistence and in the pathology progression.

Published
2008
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10. Production of biologically active forms of recombinant hepcidin, the iron-regulatory hormone

Author

Gagliardo, Bruno, Faye, Audrey, Jaouen, Maryse, Deschemin, Jean-Christophe, Canonne-Hergaux, François, Vaulont, Sophie, Sari, Marie-Agnès, Institut Pasteur de Dakar, Réseau International des Instituts Pasteur (RIIP), Service de pédiatrie générale, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 (UPD7), Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Laboratoire de métallurgie physique (LMP), Centre National de la Recherche Scientifique (CNRS)-Université de Poitiers, Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie des Substances Naturelles (ICSN), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Génétique et pathologie moléculaires, Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de génétique moléculaire (CGM), Centre National de la Recherche Scientifique (CNRS), Institut Pasteur de Dakar-Réseau International des Instituts Pasteur ( RIIP ), Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 ( UPD7 ), Institut de biologie et chimie des protéines [Lyon] ( IBCP ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de métallurgie physique ( LMP ), Université de Poitiers-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques ( LCBPT - UMR 8601 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de Chimie des Substances Naturelles ( ICSN ), Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Cochin ( UMR_S567 / UMR 8104 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Centre de génétique moléculaire ( CGM ), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 (UPD7), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)

Subjects
inorganic chemicals, MESH: DNA Primers, MESH : Recombinant Proteins, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH: Base Sequence, MESH: Iron-Regulatory Proteins, MESH : Electrophoresis, Polyacrylamide Gel, Mass Spectrometry, MESH: Recombinant Proteins, Mice, MESH : Chromatography, High Pressure Liquid, MESH : Mass Spectrometry, Hepcidins, hemic and lymphatic diseases, MESH : Mice, Animals, Humans, MESH: Animals, MESH: Chromatography, High Pressure Liquid, MESH: Mice, Chromatography, High Pressure Liquid, DNA Primers, MESH: Mass Spectrometry, MESH: Humans, Base Sequence, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, MESH : Humans, MESH: Antimicrobial Cationic Peptides, Iron-Regulatory Proteins, nutritional and metabolic diseases, MESH : Iron-Regulatory Proteins, MESH : Antimicrobial Cationic Peptides, Recombinant Proteins, Electrophoresis, Polyacrylamide Gel, MESH : Base Sequence, MESH : DNA Primers, MESH : Animals, Antimicrobial Cationic Peptides, MESH: Electrophoresis, Polyacrylamide Gel
Abstract

International audience; Hepcidin is a liver produced cysteine-rich peptide hormone that acts as the central regulator of body iron metabolism. Hepcidin is synthesized under the form of a precursor, prohepcidin, which is processed to produce the biologically active mature 25 amino acid peptide. This peptide is secreted and acts by controlling the concentration of the membrane iron exporter ferroportin on intestinal enterocytes and macrophages. Hepcidin binds to ferroportin, inducing its internalization and degradation, thus regulating the export of iron from cells to plasma. The aim of the present study was to develop a novel method to produce human and mouse recombinant hepcidins, and to compare their biological activity towards their natural receptor ferroportin. Hepcidins were expressed in Escherichia coli as thioredoxin fusion proteins. The corresponding peptides, purified after cleavage from thioredoxin, were properly folded and contained the expected four-disulfide bridges without the need of any renaturation or oxidation steps. Human and mouse hepcidins were found to be biologically active, promoting ferroportin degradation in macrophages. Importantly, biologically inactive aggregated forms of hepcidin were observed depending on purification and storage conditions, but such forms were unrelated to disulfide bridge formation.

Published
2008
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11. Diversity surveys and evolutionary relationships of aoxB genes in aerobic arsenite-oxidizing bacteria

Author

Marianne Quéméneur, Michel Jauzein, Audrey Heinrich-Salmeron, Didier Lièvremont, Catherine Joulian, Daniel Muller, Philippe N. Bertin, Francis Garrido, Génétique moléculaire, génomique, microbiologie (GMGM), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Lyon (ENVL)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Laboratoire de Biologie Tumorale, CRLCC Paul Strauss, XLIM (XLIM), Université de Limoges (UNILIM)-Centre National de la Recherche Scientifique (CNRS), Bureau de Recherches Géologiques et Minières (BRGM) (BRGM), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Ecole Nationale Vétérinaire de Lyon (ENVL), Génétique moléculaire, génomique, microbiologie ( GMGM ), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique ( CNRS ), Ecologie microbienne ( EM ), Centre National de la Recherche Scientifique ( CNRS ) -Ecole Nationale Vétérinaire de Lyon ( ENVL ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique ( INRA ) -VetAgro Sup ( VAS ), XLIM ( XLIM ), Université de Limoges ( UNILIM ) -Centre National de la Recherche Scientifique ( CNRS ), and Bureau de Recherches Géologiques et Minières (BRGM) ( BRGM )

Subjects
Chemoautotrophic Growth, MESH: Sequence Analysis, Protein, MESH: Amino Acid Sequence, MESH: Base Sequence, Polymerase Chain Reaction, MESH : Arsenites, Conserved sequence, Sequence Analysis, Protein, RNA, Ribosomal, 16S, MESH : Genetic Markers, MESH : DNA, Bacterial, Soil Pollutants, MESH: Genetic Variation, MESH: Phylogeny, MESH : Proteobacteria, Phylogeny, 0303 health sciences, MESH: Conserved Sequence, MESH : Amino Acid Sequence, MESH : Genes, Bacterial, Bacterial, MESH : Oxidoreductases, MESH: RNA, Ribosomal, 16S, MESH : DNA Primers, MESH: Arsenites, Proteobacteria, Genetic Markers, MESH: DNA Primers, 16S, Sequence analysis, Arsenites, MESH : Soil Microbiology, Molecular Sequence Data, MESH: Biodiversity, 03 medical and health sciences, Phylogenetics, MESH : Conserved Sequence, MESH : Bacteria, Amino Acid Sequence, MESH: Oxidoreductases, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Oxidation-Reduction, Ribosomal, MESH: Soil Pollutants, MESH: Molecular Sequence Data, Bacteria, 030306 microbiology, Alphaproteobacteria, Genetic Variation, MESH: Polymerase Chain Reaction, DNA, Ribosomal RNA, MESH: DNA, Bacterial, Genes, MESH : Sequence Analysis, Protein, MESH : Sequence Analysis, DNA, MESH: Oxidation-Reduction, MESH: Sequence Analysis, DNA, MESH : Molecular Sequence Data, MESH: Genetic Markers, Applied Microbiology and Biotechnology, MESH : Soil Pollutants, MESH : Biodiversity, MESH : Polymerase Chain Reaction, Betaproteobacteria, Conserved Sequence, Soil Microbiology, Genetics, Ecology, biology, Biodiversity, MESH: Genes, Bacterial, Oxidoreductases, Oxidation-Reduction, Sequence Analysis, Biotechnology, DNA, Bacterial, MESH : Chemoautotrophic Growth, Microbial Ecology, MESH : Genetic Variation, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, 030304 developmental biology, DNA Primers, Base Sequence, Protein, MESH : Phylogeny, Sequence Analysis, DNA, MESH: Proteobacteria, biology.organism_classification, 16S ribosomal RNA, MESH: Chemoautotrophic Growth, MESH : RNA, Ribosomal, 16S, MESH: Bacteria, MESH: Soil Microbiology, Genes, Bacterial, RNA, MESH : Base Sequence, Food Science
Abstract

A new primer set was designed to specifically amplify ca. 1,100 bp of aoxB genes encoding the As(III) oxidase catalytic subunit from taxonomically diverse aerobic As(III)-oxidizing bacteria. Comparative analysis of AoxB protein sequences showed variable conservation levels and highlighted the conservation of essential amino acids and structural motifs. AoxB phylogeny of pure strains showed well-discriminated taxonomic groups and was similar to 16S rRNA phylogeny. Alphaproteobacteria -, Betaproteobacteria -, and Gammaproteobacteria -related sequences were retrieved from environmental surveys, demonstrating their prevalence in mesophilic As-contaminated soils. Our study underlines the usefulness of the aoxB gene as a functional marker of aerobic As(III) oxidizers.

Published
2008
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12. MicroRNA166 controls root and nodule development in Medicago truncatula

Author

Carole Laffont, Martin Crespi, Andreas Niebel, Julie Plet, Philippe Laporte, Jean-Philippe Combier, Mariana Jovanovic, Florian Frugier, Adnane Boualem, Institut des sciences du végétal (ISV), Centre National de la Recherche Scientifique (CNRS), Nouveaux Outils pour La Coopération et l'Education (NOCE), Laboratoire d'Informatique Fondamentale de Lille (LIFL), Université de Lille, Sciences et Technologies-Institut National de Recherche en Informatique et en Automatique (Inria)-Université de Lille, Sciences Humaines et Sociales-Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Sciences et Technologies-Institut National de Recherche en Informatique et en Automatique (Inria)-Université de Lille, Sciences Humaines et Sociales-Centre National de la Recherche Scientifique (CNRS), Université de Lille, Sciences et Technologies-Institut National de Recherche en Informatique et en Automatique (Inria)-Université de Lille, Sciences Humaines et Sociales-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Ecole Nationale Vétérinaire de Lyon (ENVL), Laboratoire des interactions plantes micro-organismes (LIPM), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Université de Lyon-Université de Lyon-Ecole Nationale Vétérinaire de Lyon (ENVL)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Lyon (ENVL)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Institut des sciences du végétal ( ISV ), Centre National de la Recherche Scientifique ( CNRS ), Nouveaux Outils pour La Coopération et l'Education ( NOCE ), Laboratoire d'Informatique Fondamentale de Lille ( LIFL ), Université de Lille, Sciences et Technologies-Institut National de Recherche en Informatique et en Automatique ( Inria ) -Université de Lille, Sciences Humaines et Sociales-Centre National de la Recherche Scientifique ( CNRS ) -Université de Lille, Sciences et Technologies-Institut National de Recherche en Informatique et en Automatique ( Inria ) -Université de Lille, Sciences Humaines et Sociales-Centre National de la Recherche Scientifique ( CNRS ), Université de Lille, Sciences et Technologies-Institut National de Recherche en Informatique et en Automatique ( Inria ) -Université de Lille, Sciences Humaines et Sociales-Centre National de la Recherche Scientifique ( CNRS ), Ecologie microbienne ( EM ), Centre National de la Recherche Scientifique ( CNRS ) -Ecole Nationale Vétérinaire de Lyon ( ENVL ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique ( INRA ) -VetAgro Sup ( VAS ), Laboratoire des interactions plantes micro-organismes ( LIPMO ), and Institut National de la Recherche Agronomique ( INRA ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
0106 biological sciences, MESH : Plant Roots, MESH : Brain Chemistry, MESH: Plant Roots, clustered miRNA, Plant Science, MESH: Amino Acid Sequence, MESH: Base Sequence, 01 natural sciences, MESH : Cattle, MESH: Cattle Diseases, MESH: Animals, In Situ Hybridization, 0303 health sciences, MESH : Amino Acid Sequence, MESH : Reverse Transcriptase Polymerase Chain Reaction, MESH : In Situ Hybridization, Medicago truncatula, Cell biology, MESH : Proton-Translocating ATPases, MESH : DNA Primers, MESH: Proteolipids, MESH: DNA Primers, MESH: Mitochondria, Organogenesis, MESH: Microscopy, Electron, MESH : Electrophoresis, Polyacrylamide Gel, 03 medical and health sciences, MESH: In Situ Hybridization, microRNA, Botany, Genetics, MESH: Blotting, Northern, MESH: Mass Spectrometry, MESH : Cattle Diseases, MESH: Molecular Sequence Data, MESH : Medicago truncatula, Lateral root, HD ZIP III transcription factors, MESH : Pigments, Biological, MESH: Neuronal Ceroid-Lipofuscinoses, MESH : Brain, MESH : Amino Acids, MESH : Gene Expression Regulation, Plant, MESH: MicroRNAs, MESH: Female, MESH: Liver, [ SDV.BV ] Life Sciences [q-bio]/Vegetal Biology, MESH : Molecular Sequence Data, MESH: Amino Acids, Plant Roots, lateral root, MESH: Proton-Translocating ATPases, Gene Expression Regulation, Plant, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Female, MESH: Brain Chemistry, MESH : Pancreas, MESH: Medicago truncatula, biology, Reverse Transcriptase Polymerase Chain Reaction, MESH : Blotting, Northern, MESH: Pancreas, MESH: Lipoproteins, MESH : Kidney, MESH: Cattle, MESH : Mitochondria, Rhizobium, MESH: Pigments, Biological, Transgene, MESH : Lipoproteins, MESH: Brain, MESH : Mass Spectrometry, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, MESH: Gene Expression Regulation, Plant, Transcription factor, Gene, 030304 developmental biology, DNA Primers, Base Sequence, polycistronic miRNA, MESH : Liver, MESH : Lipids, Cell Biology, MESH: Kidney, Meristem, biology.organism_classification, Blotting, Northern, symbiotic nodulation, MESH: Lipids, MESH : Microscopy, Electron, MESH : MicroRNAs, MicroRNAs, MESH : Proteolipids, MESH : Base Sequence, MESH : Animals, MESH : Neuronal Ceroid-Lipofuscinoses, 010606 plant biology & botany, MESH: Electrophoresis, Polyacrylamide Gel
Abstract

International audience; Legume root architecture is characterized by the development of two de novo meristems, leading to the formation of lateral roots or symbiotic nitrogen-fixing nodules. Organogenesis involves networks of transcription factors, the encoding mRNAs of which are frequently targets of microRNA (miRNA) regulation. Most plant miRNAs, in contrast with animal miRNAs, are encoded as single entities in an miRNA precursor. In the model legume Medicago truncatula, we have identified the MtMIR166a precursor containing tandem copies of MIR166 in a single transcriptional unit. These miRNAs post-transcriptionally regulate a new family of transcription factors associated with nodule development, the class-III homeodomain-leucine zipper (HD-ZIP III) genes. In situ expression analysis revealed that these target genes are spatially co-expressed with MIR166 in vascular bundles, and in apical regions of roots and nodules. Overexpression of the tandem miRNA precursor correlated with MIR166 accumulation and the downregulation of several class-III HD-ZIP genes, indicating its functionality. MIR166 overexpression reduced the number of symbiotic nodules and lateral roots, and induced ectopic development of vascular bundles in these transgenic roots. Hence, plant polycistronic miRNA precursors, although rare, can be processed, and MIR166-mediated post-transcriptional regulation is a new regulatory pathway involved in the regulation of legume root architecture.

Published
2008
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13. Analysis of human papillomavirus type 16 (HPV16) DNA load and physical state for identification of HPV16-infected women with high-grade lesions or cervical carcinoma

Author

Sylvain Monnier-Benoit, Frédéric Mauny, Christiane Mougin, Jean-Luc Prétet, Didier Riethmuller, Véronique Dalstein, Elisabeth Schwarz, Jenny Briolat, Maëlle Saunier, Bernadette Kantelip, 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 ), Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Laboratoire d'anatomie pathologique [Besancon], Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Jean Minjoz-Université de Franche-Comté ( UFC ), Laboratoire Chrono-environnement - CNRS - UFC (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC (EA 3181) (CEF2P / CARCINO), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Service d'Anatomie pathologique [CHRU Besançon], and Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)

Subjects
Oncology, Pathology, Statistics as Topic, MESH : Statistics as Topic, Uterine Cervical Neoplasms, MESH: Papillomavirus Infections, Polymerase Chain Reaction, law.invention, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, chemistry.chemical_compound, 0302 clinical medicine, law, MESH : Female, MESH : DNA, Viral, MESH : Polymerase Chain Reaction, Polymerase chain reaction, MESH: Human papillomavirus 16, Cervical cancer, MESH : Papillomavirus Infections, Human papillomavirus 16, 0303 health sciences, [ SDV.SPEE ] Life Sciences [q-bio]/Santé publique et épidémiologie, MESH : Oncogene Proteins, Viral, 3. Good health, DNA-Binding Proteins, MESH: Uterine Cervical Neoplasms, MESH: Repressor Proteins, 030220 oncology & carcinogenesis, Female, MESH : DNA Primers, MESH : DNA-Binding Proteins, medicine.symptom, MESH : Repressor Proteins, Viral load, Microbiology (medical), MESH: DNA Primers, medicine.medical_specialty, MESH : Uterine Cervical Neoplasms, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, Virus, Lesion, 03 medical and health sciences, Virology, Internal medicine, medicine, Humans, Gene, MESH: Statistics as Topic, DNA Primers, 030304 developmental biology, MESH: Humans, Papillomavirus Infections, MESH : Humans, MESH : Human papillomavirus 16, Cancer, MESH: Polymerase Chain Reaction, Oncogene Proteins, Viral, medicine.disease, MESH: DNA, Viral, Repressor Proteins, chemistry, DNA, Viral, MESH: Oncogene Proteins, Viral, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, MESH: Female, DNA, MESH: DNA-Binding Proteins
Abstract

Integration of human papillomavirus (HPV) DNA into the host cell genome is a frequent event in cervical carcinogenesis, even though this phenomenon does not seem to be mandatory for cervical cancer development. Our objective was to describe the load and physical state of HPV type 16 (HPV16) DNA in a series of cervical samples representative of the natural history of cervical cancer. We used a combination of three real-time PCR assays targeting E6, E2, and albumin genes to calculate HPV16 load (E6 and albumin) and the E2/E6 ratio as a surrogate of integration. This method was applied to 173 HPV16-positive cervical samples. Results show that viral load increases with the lesion grade (from 102 HPV16 DNA copies per 10 3 cells in normal samples up to 56,354 copies per 10 3 cells in cancers), while E2/E6 ratio decreases (from 1 in normal samples down to 0.36 in cancers). We propose that, according to this technique, an HPV16 viral load of higher than 22,000 copies/10 3 cells or an E2/E6 ratio of lower than 0.50 allows the identification of women with prevalent high-grade lesions or worse with a high specificity. In conclusion, both viral load and E2/E6 ratio, used in combination with an appropriate cutoff value, are suitable to screen women with prevalent cervical intraepithelial neoplasia grade 2 or 3 or cancer. Therefore, these assays would be useful in addition to routine HPV testing to more accurately identify women with (pre)cancerous lesions.

Published
2008
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14. Glycogen synthase 2 is a novel target gene of peroxisome proliferator-activated receptors

Author

Mandard, Stéphane, Stienstra, Rinke, Escher, Pascal, Tan, Nguan Soon, Kim, Insook, Gonzalez, Frank, Wahli, Walter, Desvergne, Béatrice, Müller, Michael, Kersten, Sander, Nutrition, Metabolism and Genomics group and Nutrigenomics Consortium ( NMG ), Wageningen University and Research Centre [Wageningen] ( WUR ), Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Institute of Physiology, University of Basel ( Unibas ), Center for Integrative Genomics - Institute of Bioinformatics, Génopode ( CIG ), University of Lausanne, Laboratory of Metabolism, National Cancer Institute ( NIH ), This study was supported by the Netherlands Organization for Scientific Research (NWO), with additional support from the Dutch Diabetes Foundation, the Royal Netherlands Academy of Art and Sciences (KNAW), Wageningen Center for Food Sciences, the Center for Human Nutrigenomics, the Swiss National Science Foundation and the Human Frontier Science Program., Nutrition, Metabolism and Genomics group and Nutrigenomics Consortium (NMG), Wageningen University and Research [Wageningen] (WUR), Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), University of Basel (Unibas), Center for Integrative Genomics - Institute of Bioinformatics, Génopode (CIG), Swiss Institute of Bioinformatics [Lausanne] (SIB), Université de Lausanne (UNIL)-Université de Lausanne (UNIL), National Cancer Institute [Bethesda] (NCI-NIH), and National Institutes of Health [Bethesda] (NIH)-National Institutes of Health [Bethesda] (NIH)

Subjects
MESH: DNA Primers, MESH: Rats, glycogen synthase 2, MESH: Peroxisome Proliferator-Activated Receptors, MESH : Hepatocytes, PPRE, liver, MESH: Mice, Knockout, PPAR, MESH: Chromatin, MESH : Gene Expression Regulation, Enzymologic, MESH: Hepatocytes, MESH: RNA, MESH : Mice, MESH : RNA, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Mice, HNF4α, MESH : Polymerase Chain Reaction, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Glycogen Synthase, MESH : Chromatin, MESH : Rats, MESH: Transcription, Genetic, MESH: Gene Expression Regulation, Enzymologic, MESH : Transcription, Genetic, MESH: Polymerase Chain Reaction, MESH : Peroxisome Proliferator-Activated Receptors, adipose tissue, gene transcription, MESH : Mice, Knockout, MESH : DNA Primers, MESH : Animals, microarray, MESH : Glycogen Synthase
Abstract

International audience; Glycogen synthase 2 (Gys-2) is the ratelimiting enzyme in the storage of glycogen in liver and adipose tissue, yet little is known about regulation of Gys-2 transcription. The peroxisome proliferator-activated receptors (PPARs) are transcription factors involved in the regulation of lipid and glucose metabolism and might be hypothesized to govern glycogen synthesis as well. Here, we show that Gys-2 is a direct target gene of PPARalpha, PPARbeta/delta and PPARgamma. Expression of Gys-2 is significantly reduced in adipose tissue of PPARalpha-/-, PPARbeta/delta-/- and PPARgamma+/- mice. Furthermore, synthetic PPARbeta/delta, and gamma agonists markedly up-regulate Gys-2 mRNA and protein expression in mouse 3T3-L1 adipocytes. In liver, PPARalpha deletion leads to decreased glycogen levels in the refed state, which is paralleled by decreased expression of Gys-2 in fasted and refed state. Two putative PPAR response elements (PPREs) were identified in the mouse Gys-2 gene: one in the upstream promoter (DR-1prom) and one in intron 1 (DR-1int). It is shown that DR-1int is the response element for PPARs, while DR-1prom is the response element for Hepatic Nuclear Factor 4 alpha (HNF4alpha). In adipose tissue, which does not express HNF4alpha, DR-1prom is occupied by PPARbeta/delta and PPARgamma, yet binding does not translate into transcriptional activation of Gys-2. Overall, we conclude that mouse Gys-2 is a novel PPAR target gene and that transactivation by PPARs and HNF4alpha is mediated by two distinct response elements.

Published
2007
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15. A PCR for the detection of mycoplasmas belonging to the Mycoplasma mycoides cluster : application to the diagnosis of contagious agalactia

Author

Lucia Manso-Silvan, François Poumarat, Marie-Pierre Pellet, François Thiaucourt, Patrice Gaurivaud, Sophie Lorenzon, V.P. Singh, Salah Woubit, Contrôle des maladies animales exotiques et émergentes ( Contrôle des maladies ), Centre de Coopération Internationale en Recherche Agronomique pour le Développement ( CIRAD ), Mycoplasmoses des ruminants [Lyon], Université de Lyon-VetAgro Sup ( VAS ), Agence Française de Sécurité Sanitaire des Aliments ( AFSSA ), AFSSA, IVRI, National Referral Lab on Mycoplasma, Contrôle des maladies animales exotiques et émergentes (Cirad-EMVT-UPR 15 Contrôle des maladies), Département Elevage et médecine vétérinaire (EMVT), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Mycoplasmoses des Ruminants - UMR (MYCO), Université de Lyon-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Agence Française de Sécurité Sanitaire des Aliments (AFSSA), and Département Elevage et médecine vétérinaire (Cirad-EMVT)

Subjects
MESH : Molecular Sequence Data, MESH : Operon, Hydrolases, Mycoplasma agalactiae, ved/biology.organism_classification_rank.species, Pcr cloning, MESH: Base Sequence, [ SDV.MP.BAC ] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, medicine.disease_cause, Polymerase Chain Reaction, Mycoplasma capricolum, Serology, 0403 veterinary science, MESH : Hydrolases, MESH: Animals, MESH: Phylogeny, MESH: Pleuropneumonia, Contagious, MESH : Polymerase Chain Reaction, Base Pairing, Phylogeny, 0303 health sciences, biology, XUMYCOPLASME, MESH : Sequence Alignment, Goats, Mycoplasma mycoides, [ SDV.SPEE ] Life Sciences [q-bio]/Santé publique et épidémiologie, 04 agricultural and veterinary sciences, 3. Good health, MESH: Hydrolases, MESH: Reproducibility of Results, MESH : Pleuropneumonia, Contagious, Milk, MESH: Mycoplasma mycoides, MESH : DNA Primers, MESH: DNA Primers, MESH: Operon, 040301 veterinary sciences, MESH: Base Pairing, Molecular Sequence Data, MESH: Sequence Alignment, MESH: Goats, MYCOPLASME, Microbiology, 03 medical and health sciences, Operon, medicine, Animals, L50 - Physiologie et biochimie animales, Pleuropneumonia, Contagious, Molecular Biology, Gene, DNA Primers, MESH: Molecular Sequence Data, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Base Sequence, 030306 microbiology, ved/biology, MESH : Reproducibility of Results, MESH : Phylogeny, Reproducibility of Results, MESH: Polymerase Chain Reaction, Cell Biology, Mycoplasma, biology.organism_classification, MESH : Base Pairing, Virology, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: Milk, MESH : Goats, MESH : Milk, HAFSSA, Mycoplasma putrefaciens, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, MESH : Base Sequence, MESH : Animals, [ SDV.BA.MVSA ] Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, MESH : Mycoplasma mycoides, Sequence Alignment
Abstract

Contagious agalactia is a mycoplasmal infection caused by Mycoplasma agalactiae, Mycoplasma mycoides subsp. mycoides LC, M. mycoides subsp. capri, Mycoplasma capricolum subsp. capricolum and Mycoplasma putrefaciens. Identification of the causative organisms is usually performed by isolation and classical biochemical and serological tests, though this is a lengthy and cumbersome process for mycoplasmas. Specific PCR assays have been developed for the identification of Mycoplasma agalactiae and M. putrefaciens. For members of the M. mycoides cluster existing PCR tests are based on the amplification of highly conserved genes coding for ribosomal proteins, hence a possibility of cross-reactions. The gene glk, coding for a glucokinase, that is found in this cluster is very distantly related to any other bacterial glucokinase described so far. It was therefore chosen as target to design a new PCR test. The validation was performed independently in three laboratories in France and India using over 100 mycoplasma strains of various geographical origins. All strains belonging to the M. mycoides cluster were detected by amplification of the expected PCR product (428 bp) while no amplification was obtained from M. agalactiae strains. Our results demonstrate the universality of this PCR in spite of the great heterogeneity found within this cluster. This new tool may be of great help for the implementation of control measures directed towards contagious agalactia. © 2007 Elsevier Ltd All rights reserved.

Published
2007
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16. Use of differentiating embryonic stem cells in pharmacological studies

Author

Wdziekonski, B., Villageois, P., Vernochet, C., Phillips, B., Christian Dani, Institut de signalisation, biologie du développement et cancer ( ISBDC ), Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS ), Institut de signalisation, biologie du développement et cancer (ISBDC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA)

Subjects
Cell Culture Techniques, Drug Evaluation, Preclinical, Gene Expression, MESH : Adipocytes, MESH: Base Sequence, Mice, Osteogenesis, MESH: Reverse Transcriptase Polymerase Chain Reaction, Adipocytes, MESH : Osteogenesis, MESH : RNA, MESH : Embryo, Mammalian, MESH: Animals, MESH : Drug Evaluation, Preclinical, MESH: Osteogenesis, [SDV.BDD]Life Sciences [q-bio]/Development Biology, Adipogenesis, Reverse Transcriptase Polymerase Chain Reaction, MESH : Adipogenesis, MESH : Reverse Transcriptase Polymerase Chain Reaction, Cell Differentiation, MESH : Pluripotent Stem Cells, MESH: Pluripotent Stem Cells, MESH: Drug Evaluation, Preclinical, MESH : DNA Primers, MESH : Cell Differentiation, Pluripotent Stem Cells, MESH: Cell Differentiation, MESH: DNA Primers, MESH : Cell Culture Techniques, MESH: Gene Expression, MESH: RNA, MESH : Mice, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, [ SDV.BDD ] Life Sciences [q-bio]/Development Biology, MESH: Mice, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Adipocytes, MESH : Osteoblasts, DNA Primers, MESH: Osteoblasts, MESH: Cell Culture Techniques, Osteoblasts, Base Sequence, MESH: Embryo, Mammalian, Embryo, Mammalian, MESH : Gene Expression, RNA, MESH : Base Sequence, MESH : Animals, MESH: Adipogenesis
Abstract

International audience; Adipocytes and osteoblasts are derived from a common precursor cell. It has been proposed that the bone loss commonly seen during aging or in the pathology of osteoporosis might be partly caused by a deregulation of the normal balance between osteoblast and adipocyte differentiation. In vitro differentiation of mouse embryonic stem cells toward the adipogenic and the osteogenic lineages provides a powerful system for testing effects of compounds on the developmental switch between adipogenesis and osteogenesis and identification of pharmacological targets. In this chapter, an improved protocol to commit mouse embryonic stem cells into both lineages at a correct rate is detailed.

Published
2006
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17. Mastocytosis in mice expressing human Kit receptor with the activating Asp816Val mutation

Author

Roland S. Liblau, Jacques P. Zappulla, Sébastien Letard, Michel Arock, Patrice Dubreuil, Nadine Ben Hamouda, Marc Daëron, Georges Delsol, Sabine Desbois, Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Cancérologie (Inserm U599/IPC), Université de la Méditerranée - Aix-Marseille 2-Institut Paoli-Calmettes, Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Fédération nationale des Centres de lutte contre le Cancer (FNCLCC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Cytokines, hématopoïèse et réponse immune (CHRI), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Allergologie Moléculaire et Cellulaire, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d'immunologie, Hôpital de Rangueil, CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse], Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Département Immunologie [CHU Toulouse], Institut Fédératif de Biologie (IFB), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Pôle Biologie [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Slama, Catherine, Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de Physiopathologie Toulouse Purpan (ex IFR 30 et IFR 150), Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IFR30-IFR150-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Cancérologie, Université de la Méditerranée - Aix-Marseille 2-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Cytokines, hématopoïèse et réponse immune ( CHRI ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), and Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM )

Subjects
Mutant, Cell Culture Techniques, MESH : Blotting, Western, medicine.disease_cause, Mice, 0302 clinical medicine, MESH: Reverse Transcriptase Polymerase Chain Reaction, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, Immunology and Allergy, MESH: Animals, Mast Cells, 0303 health sciences, Mutation, Reverse Transcriptase Polymerase Chain Reaction, MESH : Reverse Transcriptase Polymerase Chain Reaction, MESH: Mast Cells, Hyperplasia, MESH : Mice, Transgenic, 3. Good health, MESH: Proto-Oncogene Proteins c-kit, Proto-Oncogene Proteins c-kit, 030220 oncology & carcinogenesis, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH : DNA Primers, MESH : Mast Cells, Mastocytosis, Genetically modified mouse, MESH: DNA Primers, MESH : Cell Culture Techniques, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Mice, Transgenic, Transgene, Blotting, Western, Immunology, Mutation, Missense, Mice, Transgenic, MESH : Mastocytosis, Biology, MESH : Proto-Oncogene Proteins c-kit, 03 medical and health sciences, MESH : Mice, medicine, Animals, Humans, MESH: Blotting, Western, MESH: Mice, DNA Primers, 030304 developmental biology, MESH: Cell Culture Techniques, MESH: Mutation, Missense, MESH: Humans, Point mutation, MESH : Humans, Brief Definitive Report, medicine.disease, Mast cell sarcoma, Cancer research, MESH: Mastocytosis, MESH : Animals, MESH : Mutation, Missense
Abstract

Mastocytosis is a rare neoplastic disease characterized by a pathologic accumulation of tissue mast cells (MCs). Mastocytosis is often associated with a somatic point mutation in the Kit protooncogene leading to an Asp/Val substitution at position 816 in the kinase domain of this receptor. The contribution of this mutation to mastocytosis development remains unclear. In addition, the clinical heterogeneity presented by mastocytosis patients carrying the same mutation is unexplained. We report that a disease with striking similarities to human mastocytosis develops spontaneously in transgenic mice expressing the human Asp816Val mutant Kit protooncogene specifically in MCs. This disease is characterized by clinical signs ranging from a localized and indolent MC hyperplasia to an invasive MC tumor. In addition, bone marrow–derived MCs from transgenic animals can be maintained in culture for >24 mo and acquire growth factor independency for proliferation. These results demonstrate a causal link in vivo between the Asp816Val Kit mutation and MC neoplasia and suggest a basis for the clinical heterogeneity of human mastocytosis.

Published
2005
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18. NMR-based identification of peptides that specifically recognize the d-arm of tRNA

Author

Frédéric Dardel, Florence Guillière, Carine Tisné, Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), and Université Paris Descartes - Paris 5 (UPD5) - Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH : Nucleic Acid Conformation, MESH: Reverse Transcription, D arm, Ligands, 01 natural sciences, Biochemistry, MESH: HIV-1, RNA, Transfer, MESH: Nuclear Magnetic Resonance, Biomolecular, MESH: Ligands, Nucleotide, Genetics, chemistry.chemical_classification, 0303 health sciences, MESH : Ligands, MESH : Peptides, MESH: Peptides, Nucleic acid sequence, General Medicine, 3. Good health, MESH: Nucleic Acid Conformation, Transfer RNA, MESH : DNA Primers, MESH : HIV-1, MESH: DNA Primers, MESH : RNA, Transfer, Biology, MESH : Nuclear Magnetic Resonance, Biomolecular, 03 medical and health sciences, MESH : Peptide Library, Peptide Library, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Peptide library, Nuclear Magnetic Resonance, Biomolecular, 030304 developmental biology, DNA Primers, Binding Sites, MESH: Humans, 010405 organic chemistry, MESH : Humans, RNA, Reverse Transcription, MESH: RNA, Transfer, Reverse transcriptase, 0104 chemical sciences, MESH : Reverse Transcription, chemistry, MESH: Binding Sites, HIV-1, Nucleic Acid Conformation, MESH: Peptide Library, Primer (molecular biology), Peptides, MESH : Binding Sites
Abstract

Human tRNA 3 Lys is used by HIV virus as a primer for the reverse transcription of its genome. The 18 nucleotides at the 3′-end of the tRNA 3 Lys are hybridized to a complementary sequence of the viral RNA called the primer-binding site. A screen against the human tRNA 3 Lys over a peptide library designed to target RNA has been performed. Of the 175 hexapeptides tested, three were found to bind to the d -stem of tRNA 3 Lys . Alanine-scanning was used to define the determinants of the interaction between the peptides and tRNA 3 Lys . They also bind to two other tested tRNAs, also at the level of the d -stem and loop, although the nucleotide sequence of the stem differs in one of them. These short peptides thus recognize specific structural features within the d -stem and loop of tRNAs. Associated with other pharmacophores, they could be useful to design optimized ligands targeting specific tRNAs such as retroviral replication primers.

Published
2005
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19. Comparative transductions of breast cancer cells by three DNA viruses

Author

Françoise Vignon, Eric J. Kremer, Silvio Hemmi, José Luis, Annick Lucas, Gwendal Lazennec, Endocrinologie moléculaire et cellulaire des cancers, Université Montpellier 1 ( UM1 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), Molecular Biology I, University of Zürich [Zürich] ( UZH ), Cytosquelette et Intégration des Signaux du Micro-Environnement Tumoral - UMR 6032 ( CISMET ), Université de la Méditerranée - Aix-Marseille 2-Université de Provence - Aix-Marseille 1-Centre National de la Recherche Scientifique ( CNRS ), Lazennec, Gwendal, Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Universität Zürich [Zürich] = University of Zurich (UZH), Cytosquelette et Intégration des Signaux du Micro-Environnement Tumoral - UMR 6032 (CISMET), Université de la Méditerranée - Aix-Marseille 2-Université de Provence - Aix-Marseille 1-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), and Centre National de la Recherche Scientifique (CNRS)-Université de Provence - Aix-Marseille 1-Université de la Méditerranée - Aix-Marseille 2

Subjects
Serotype, MESH: DNA Primers, viruses, Biophysics, [SDV.CAN]Life Sciences [q-bio]/Cancer, Breast Neoplasms, MESH : Transduction, Genetic, MESH: Base Sequence, MESH : Breast Neoplasms, medicine.disease_cause, Biochemistry, Virus, Flow cytometry, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, 03 medical and health sciences, 0302 clinical medicine, Breast cancer, [SDV.CAN] Life Sciences [q-bio]/Cancer, Transduction, Genetic, medicine, Humans, Vector (molecular biology), Molecular Biology, Adeno-associated virus, 030304 developmental biology, DNA Primers, 0303 health sciences, MESH: Humans, biology, medicine.diagnostic_test, Base Sequence, MESH : Humans, DNA Viruses, Cancer, Cell Biology, medicine.disease, MESH: Transduction, Genetic, Virology, MESH: DNA Viruses, 3. Good health, 030220 oncology & carcinogenesis, biology.protein, MESH : DNA Viruses, MESH : Base Sequence, MESH : DNA Primers, Antibody, MESH: Breast Neoplasms
Abstract

Defining the ideal vectors to transduce breast cancer using viruses is currently under intense pre-clinical evaluation. Our study constitutes the first direct comparison of the infection efficiencies of a human serotype 5 (Ad5), a canine serotype 2 (CAV-2) adenovirus, and a human serotype 2 adeno-associated virus (AAV-2) in breast cancer cells. We observed an excellent infection efficiency for Ad5 vector, whereas both CAV-2 and AAV-2 vectors lead to low infection of these cells. Real-time PCR, flow cytometry, and antibody blocking studies suggest that Ad5 and CAV-2 infection ability is not strictly dependent on coxsackie adenovirus receptor (CAR) or alpha(v) integrin levels. In conclusion, our data suggest that human adenoviruses are excellent transducers of breast cancer cells, though it may be difficult to predict the extent of infection solely on CAR or alpha(v) integrin levels.

Published
2003
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20. Truncation of C-mip (Tc-mip), a new proximal signaling protein, induces c-maf Th2 transcription factor and cytoskeleton reorganization

Author

Sabine Le gouvelo, Asta Valanciuté, Philippe Grimbert, Georges Guellaen, Albert Bensman, Djillali Sahali, André Pawlak, Philippe Lang, Vincent Audard, Patrick Niaudet, Régulation des gènes et signalisation cellulaire, Institut National de la Santé et de la Recherche Médicale ( INSERM ), Service de néphrologie et transplantation, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ), Service de néphrologie pédiatrique [CHU Necker], Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Service de néphrologie et pédiatrie générale, CHU Trousseau [APHP]-Assistance publique - Hôpitaux de Paris (AP-HP)-Université Pierre et Marie Curie - Paris 6 ( UPMC ), Institut National de la Santé et de la Recherche Médicale (INSERM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service de néphrologie et pédiatrie générale [CHU Trousseau], Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), and Maillochon, Franck

Subjects
[ SDV.MHEP.UN ] Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology, Nephrotic Syndrome, MESH: Cytoskeletal Proteins, T-Lymphocytes, 030232 urology & nephrology, MESH: Base Sequence, Polymerase Chain Reaction, Jurkat cells, [SDV.MHEP.UN]Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology, MESH: Recombinant Proteins, Jurkat Cells, MESH : Cytoskeleton, 0302 clinical medicine, MESH : Child, T-Lymphocyte Subsets, MESH: Child, MESH: Jurkat Cells, Immunology and Allergy, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, Interferon gamma, Child, MESH : Polymerase Chain Reaction, Cytoskeleton, MESH : Jurkat Cells, Sequence Deletion, 0303 health sciences, Reverse Transcriptase Polymerase Chain Reaction, Kinase, MESH : Adult, Recombinant Proteins, DNA-Binding Proteins, Pleckstrin homology domain, medicine.anatomical_structure, lipoid nephrosis, Proto-Oncogene Proteins c-maf, MESH : Proto-Oncogene Proteins, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH : DNA Primers, MESH : DNA-Binding Proteins, MESH: Proto-Oncogene Proteins c-maf, signal transduction, medicine.drug, Proto-oncogene tyrosine-protein kinase Src, Adult, MESH: DNA Primers, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH : Recombinant Proteins, T cell, MESH : Nephrotic Syndrome, Immunology, T lymphocytes, MESH : Researc, MESH : Proto-Oncogene Proteins c-maf, Biology, Transfection, Article, src Homology Domains, 03 medical and health sciences, Th2 Cells, Proto-Oncogene Proteins, medicine, MESH: Cytoskeleton, Humans, Transcription factor, Interleukin 4, Adaptor Proteins, Signal Transducing, DNA Primers, 030304 developmental biology, pleckstrin domain, MESH: Humans, Base Sequence, MESH : Humans, MESH: Researc, MESH: Adult, MESH: Polymerase Chain Reaction, MESH : Cytoskeletal Proteins, Molecular biology, [SDV.MHEP.UN] Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology, MESH: Proto-Oncogene Proteins, Cytoskeletal Proteins, MESH : Base Sequence, MESH: Nephrotic Syndrome, MESH: DNA-Binding Proteins, Transcription Factors
Abstract

Several arguments suggest that minimal change nephrotic syndrome (MCNS) results from yet unknown systemic disorder of T cell function. By screening a cDNA library from T cell relapse, we identified a new pleckstrin homology (PH) domain-containing protein encoded by a gene located on chromosome 16q24. Two alternative transcripts were identified. The first species (c-mip) was expressed in fetal liver, kidney, and peripheral blood mononuclear cells (PBMCs), but weakly detected in PBMCs from MCNS patients. The second form (Tc-mip, standing for truncated c-maf inducing protein), corresponds to subtracted transcript and lacks the NH2-terminal PH domain. The expression of Tc-mip was restricted to fetal liver, thymus, and MCNS PBMCs where it was specifically recruited in CD4+ T cells subset. Overexpression of Tc-mip in T cell Jurkat induced c-maf, transactivated the interleukin 4 gene and down-regulated the interferon γ expression, characteristic of a Th2 commitment. Moreover, the overexpression of Tc-mip induced Src phosphorylation, T cell clustering, and a cellular redistribution of the cytoskeleton-associated L-plastin, by a PI3 kinase independent pathway. Tc-mip represents therefore the first identified protein, which links proximal signaling to c-maf induction.

Published
2003
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21. Interleukin-1beta induces glycosaminoglycan synthesis via the prostaglandin E2 pathway in cultured human cervical fibroblasts

Author

Michelle Breuiller-Fouché, Françoise Ferré, Thomas Schmitz, Dominique Cabrol, Emmanuelle Dallot, Marie-Josèphe Leroy, Breuiller-Fouche, Michelle, La grossesse normale et pathologique: développement et fonctions du placenta et de l'utérus (UMR_S 767), Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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 de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Maternité Port-Royal [CHU Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Génomique et épigénétique des pathologies placentaires (Inserm U709), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5), La grossesse normale et pathologique: développement et fonctions du placenta et de l'utérus ( UMR_S 767 ), Institut des sciences du Médicament -Toxicologie - Chimie - Environnement ( IFR71 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP], Génomique et épigénétique des pathologies placentaires ( Inserm U709 ), and Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Paris Descartes - Paris 5 ( UPD5 )

Subjects
MESH : RNA, Messenger, Transcription, Genetic, medicine.medical_treatment, MESH: Sulfonamides, Cervix Uteri, MESH: Base Sequence, 0302 clinical medicine, Cells, Cultured, Glycosaminoglycans, MESH : Isoenzymes, Sulfonamides, 0303 health sciences, 030219 obstetrics & reproductive medicine, MESH: Dinoprostone, MESH: Cervix Uteri, Receptors, Prostaglandin E, EP3 Subtype, MESH : Biphenyl Compounds, MESH : DNA Primers, MESH: Membrane Proteins, Prostaglandin E, MESH: Cells, Cultured, MESH: DNA Primers, medicine.medical_specialty, MESH : Cyclooxygenase 2, MESH: Prostaglandin-Endoperoxide Synthases, Dinoprostone, Article, 03 medical and health sciences, MESH: Cyclooxygenase Inhibitors, MESH : Fibroblasts, Genetics, Humans, Cyclooxygenase Inhibitors, RNA, Messenger, Molecular Biology, Nitrobenzenes, MESH: Humans, Cyclooxygenase 2 Inhibitors, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, Biphenyl Compounds, MESH : Humans, MESH: Interleukin-1, Fibroblasts, Receptors, Prostaglandin E, EP2 Subtype, MESH : Nitrobenzenes, Endocrinology, Reproductive Medicine, MESH : Membrane Proteins, MESH: Female, Developmental Biology, Embryology, MESH: Cyclooxygenase 2 Inhibitors, MESH: Receptors, Prostaglandin E, MESH : Cyclooxygenase 2 Inhibitors, MESH : Female, Prostaglandin E2, Receptor, MESH : Prostaglandin-Endoperoxide Synthases, MESH: Kinetics, Obstetrics and Gynecology, Interleukin, MESH: Glycosaminoglycans, MESH : Cyclooxygenase Inhibitors, Receptor antagonist, Receptors, Prostaglandin E, EP1 Subtype, Isoenzymes, MESH: Isoenzymes, lipids (amino acids, peptides, and proteins), Female, Signal transduction, MESH : Kinetics, MESH: Cyclooxygenase 2, MESH : Receptors, Prostaglandin E, MESH: Biphenyl Compounds, medicine.drug, MESH : Dinoprostone, medicine.drug_class, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, MESH : Interleukin-1, Paracrine signalling, Internal medicine, MESH : Cells, Cultured, medicine, MESH : Glycosaminoglycans, Receptors, Prostaglandin E, Autocrine signalling, MESH : Sulfonamides, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, DNA Primers, 030304 developmental biology, MESH: RNA, Messenger, Base Sequence, MESH: Transcription, Genetic, Membrane Proteins, MESH : Transcription, Genetic, MESH: Nitrobenzenes, Cell Biology, Kinetics, Cyclooxygenase 2, Prostaglandin-Endoperoxide Synthases, MESH: Fibroblasts, MESH : Cervix Uteri, MESH : Base Sequence, Receptors, Prostaglandin E, EP4 Subtype, Interleukin-1
Abstract

International audience; The aim of this study was to identify, in cultured human cervical fibroblasts, the mechanisms by which interleukin (IL)-1beta induces the synthesis of glycosaminoglycans (GAG) and to explore the putative role of prostaglandin E(2) (PGE(2)) in this process. Exposure of the cells for 24 h to IL-1beta induced a significant (P < 0.05) dose-dependent increase in GAG synthesis. IL-1beta (1 ng/ml) induced the expression of cyclooxygenase-2 (COX-2) protein 6 h after treatment, accompanied by a 7.5-fold increase in PGE(2) production. We confirmed that NS398, a selective COX-2 inhibitor, dose-dependently blocked PGE(2) augmentation following IL-1beta treatment. AH23848, the selective EP(4) receptor antagonist, completely abolished IL-1beta-induced GAG synthesis, whereas AH6809, an EP(2) receptor antagonist, had no effect on the stimulatory effects of IL-1beta. Furthermore, we demonstrated that 6 h exposure to IL-1beta induced a notable increase in EP(4) receptor mRNA expression and a decrease in EP(1) receptor mRNA but had no effect on the expression of EP(2) and EP(3) receptor transcripts. In conclusion, these findings indicate that IL-1beta not only induced GAG synthesis by increasing COX-2 protein expression and the subsequent PGE(2) production but also enhanced the responsiveness of cervical fibroblasts to PGE(2) by selectively up-regulating EP(4) receptor mRNA expression. These results suggest that PGE(2) may regulate human cervical ripening in an autocrine/paracrine manner via EP(4) receptors.

Published
2003

22. Q/R editing of the rat GluR5 and GluR6 kainate receptors in vivo and in vitro: evidence for independent developmental, pathological and cellular regulation

Author

A, Bernard, L, Ferhat, F, Dessi, G, Charton, A, Represa, Y, Ben-Ari, M, Khrestchatisky, Tyzio, Roman, Epilepsie et Ischemie Cerebrale, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM )

Subjects
Male, MESH : RNA, Messenger, MESH: Hippocampus, MESH : Cerebellum, MESH: Neurons, MESH : Hippocampus, MESH : Receptors, Kainic Acid, Hippocampus, MESH: Pregnancy, Receptors, Kainic Acid, Pregnancy, MESH : Glutamic Acid, Cerebellum, MESH: Gene Expression Regulation, Developmental, Excitatory Amino Acid Agonists, MESH : Female, MESH: Animals, MESH: Receptors, Kainic Acid, MESH : Olfactory Bulb, Cells, Cultured, Cerebral Cortex, Neurons, Kainic Acid, MESH : Rats, MESH : Neuroglia, MESH : Epilepsy, Age Factors, Gene Expression Regulation, Developmental, MESH: Glutamic Acid, Olfactory Bulb, MESH: Epilepsy, MESH : Cerebral Cortex, Female, MESH : DNA Primers, MESH: Neuroglia, Neuroglia, MESH: Cells, Cultured, MESH: Olfactory Bulb, MESH: DNA Primers, MESH: Rats, MESH : Male, Glutamic Acid, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, MESH : Rats, Wistar, MESH : Neurons, MESH : Cells, Cultured, MESH : RNA Editing, Animals, MESH: RNA Editing, RNA, Messenger, Rats, Wistar, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, DNA Primers, MESH: RNA, Messenger, MESH : Excitatory Amino Acid Agonists, MESH: Age Factors, Epilepsy, [ SDV.BC ] Life Sciences [q-bio]/Cellular Biology, MESH: Rats, Wistar, MESH: Kainic Acid, MESH: Male, MESH: Cerebellum, MESH: Cerebral Cortex, Rats, MESH : Kainic Acid, MESH : Pregnancy, MESH : Potassium, MESH: Potassium, Potassium, RNA Editing, MESH : Age Factors, MESH : Animals, MESH : Gene Expression Regulation, Developmental, MESH: Excitatory Amino Acid Agonists, MESH: Female
Abstract

International audience; Kainate (KA) is a potent neuroexcitatory agent in several areas of the adult brain, with convulsant and excitotoxic properties that increase as ontogeny proceeds. Besides its depolarizing actions, KA may enhance intracellular accumulation of Ca2+ to promote selective neuronal damage. The effects of KA are mediated by specific receptors recently considered to be involved in fast neurotransmission and that can be activated synaptically. KA receptors, e.g. GluR5 and GluR6 have been characterized by molecular cloning. Structure-function relationships indicate that in the MII domain of these KA receptors, a glutamine (Q) or arginine (R) residue determines ion selectivity. The arginine stems from post-transcriptional editing of the GluR5 and GluR6 pre-RNAs, and the unedited and edited versions of GluR6 elicit distinct Ca2+ permeability. Using a PCR-based approach, we show that in vivo, Q/R editing in the GluR5 and GluR6 mRNAs is modulated during ontogeny and differs substantially in a variety of nervous tissues. GluR5 editing is highest in peripheral nervous tissue, e.g. the dorsal root ganglia, where GluR6 expression is barely detectable. In contrast, GluR6 editing is maximal in forebrain and cerebellar structures where GluR5 editing is lower. Intra-amygdaloid injections of KA provide a model of temporal lobe epilepsy, and we show that following seizures, the extent of GluR5 and GluR6 editing is altered in the hippocampus. However, in vitro, high levels of glutamate and potassium-induced depolarizations have no effect on GluR5 and GluR6 Q/R editing. GluR6 editing is rapidly enhanced to maximal levels in primary cultures of cerebellar granule neurons but not in cultured hippocampal pyramidal neurons. Finally, we show that cultured glial cells express partially edited GluR6 mRNAs. Our results indicate that Q/R editing of GluR5 and GluR6 mRNAs is structure-, cell type- and time-dependent, and suggest that editing of these mRNAs is not co-regulated.

Published
1999

23. Introduced Siberian Chipmunks (Tamias sibiricus barberi) Contribute More to Lyme Borreliosis Risk than Native Reservoir Rodents

Author

Benoît Pisanu, Patrick Gasqui, Elisabeth Ferquel, Jean-Louis Chapuis, Sébastien Masséglia, Anne Dozières, Maud Marsot, Gwenaël Vourc’h, Centre d'Ecologie et des Sciences de la COnservation (CESCO), Muséum national d'Histoire naturelle (MNHN)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherches sur les Herbivores - UMR 1213 (UMRH), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Recherche Agronomique (INRA), Centre National de Référence des Borrelia (CNR), Institut Pasteur [Paris], Unité d'épidémiologie animale, Institut National de la Recherche Agronomique (INRA), Institut Pasteur [Paris] (IP), Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Muséum national d'Histoire naturelle (MNHN), Centre d'Ecologie et des Sciences de la COnservation ( CESCO ), Centre National de la Recherche Scientifique ( CNRS ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Muséum National d'Histoire Naturelle ( MNHN ), Unité Mixte de Recherches sur les Herbivores ( UMR 1213 Herbivores ), VetAgro Sup ( VAS ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de la Recherche Agronomique ( INRA ), and Centre National de Référence des Borrelia ( CNR )

Subjects
0106 biological sciences, Disease reservoir, MESH : Prevalence, Epidemiology, BURGDORFERI SENSU-LATO, Polymerase Chain Reaction, 01 natural sciences, Mice, MESH : Murinae, RNA, Ribosomal, 16S, MESH: Animals, lcsh:Science, Ground squirrel, 0303 health sciences, Ecology, ASSOCIATION, MESH: RNA, Ribosomal, 16S, MESH: Introduced Species, Medicine, MESH : DNA Primers, MESH: Tick Infestations, MESH: DNA Primers, EUROPE, Ixodes ricinus, MESH: Population Density, MESH: Murinae, Tick, Microbiology, MESH: Ixodes, 03 medical and health sciences, MESH : Population Density, Biology, MESH: Prevalence, MESH : Borrelia burgdorferi, MESH: Disease Reservoirs, [ SDV ] Life Sciences [q-bio], Ixodes, 030306 microbiology, lcsh:R, IXODES-RICINUS TICKS, Vectors and Hosts, MESH: Polymerase Chain Reaction, MESH: Arvicolinae, bacterial infections and mycoses, medicine.disease, MESH : Ixodes, Borrelia burgdorferi, MESH : Tick Infestations, Veterinary Science, lcsh:Q, DISEASE SPIROCHETE, DIFFERENTIAL TRANSMISSION, FRANCE, HOSTS, DIVERSITY, INFECTION, Bacterial Diseases, MESH: Sciuridae, MESH: Lyme Disease, [SDV]Life Sciences [q-bio], lcsh:Medicine, Wildlife, Ticks, Lyme disease, Zoonoses, MESH : Disease Reservoirs, Prevalence, MESH : Polymerase Chain Reaction, MESH : Lyme Disease, Lyme Disease, Multidisciplinary, biology, Arvicolinae, MESH : Sciuridae, MESH : Introduced Species, Sciuridae, Biodiversity, Siberian chipmunk, Infectious Diseases, MESH: Borrelia burgdorferi, France, Research Article, Animal Types, 010603 evolutionary biology, Infectious Disease Epidemiology, Veterinary Epidemiology, MESH : Mice, parasitic diseases, medicine, Animals, MESH : France, MESH: Mice, DNA Primers, Disease Reservoirs, Population Density, biology.organism_classification, Borrelia Infection, Tick Infestations, MESH : Arvicolinae, MESH : RNA, Ribosomal, 16S, MESH: France, Emerging Infectious Diseases, MESH : Animals, Murinae, Introduced Species
Abstract

International audience; The variation of the composition in species of host communities can modify the risk of disease transmission. In particular, the introduction of a new host species can increase health threats by adding a new reservoir and/or by amplifying the circulation of either exotic or native pathogens. Lyme borreliosis is a multi-host vector-borne disease caused by bacteria belonging to the Borrelia burgdorferi sensu lato complex. It is transmitted by the bite of hard ticks, especially Ixodes ricinus in Europe. Previous studies showed that the Siberian chipmunk, Tamias sibiricus barberi, an introduced ground squirrel in the Forest of Sénart (near Paris, France) was highly infested by I. ricinus, and consequently infected by B. burgdorferi sl. An index of the contribution of chipmunks to the density of infected questing nymphs on the vegetation (i.e., the acarological risk for humans) was compared to that of bank voles (Myodes glareolus) and of wood mice (Apodemus sylvaticus), two known native and sympatric competent reservoir hosts. Chipmunks produced nearly 8.5 times more infected questing nymphs than voles and mice. Furthermore, they contribute to a higher diversity of B. burgdorferi sl genospecies (B. afzelii, B. burgdorferi sensu stricto and B. garinii). The contribution of chipmunks varied between years and seasons, according to tick availability. As T. s. barberi must be a competent reservoir, it should amplify B. burgdorferi sl infection, hence increasing the risk of Lyme borreliosis in humans.

Published
2013
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24. Persistent Wolbachia and Cultivable Bacteria Infection in the Reproductive and Somatic Tissues of the Mosquito Vector Aedes albopictus

Author

Karima Zouache, Laurence Mousson, Van Tran-Van, Denis Voronin, Anna-Bella Failloux, Patrick Mavingui, Laboratoire d'Ecologie Microbienne - UMR 5557 (LEM), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Ecole Nationale Vétérinaire de Lyon (ENVL), Génétique Moléculaire des Bunyavirus, Institut Pasteur [Paris], KZ was supported by a PhD fellowship from the French Ministère de l'Education Nationale, de la Recherche et des Nouvelles Technologies. This work was funded by the CNRS and the Agence Nationale de Recherche (ChikVendoM ANR-06-SEST07). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., ANR-06-SEST-0007,CHIKVENDOM,Interactions multipartites entre le virus chikungunya, les endosymbiostes et les moustiques. Impact sur la transmission virale et la dynamique des populations vectorielles(2006), Ecologie microbienne ( EM ), Centre National de la Recherche Scientifique ( CNRS ) -Ecole Nationale Vétérinaire de Lyon ( ENVL ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique ( INRA ) -VetAgro Sup ( VAS ), ANR-06-SEST-0007,CHIKVENDOM,Interactions multipartites entre le virus chikungunya, les endosymbiostes et les moustiques. Impact sur la transmission virale et la dynamique des populations vectorielles ( 2006 ), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Lyon (ENVL)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Université de Lyon-Université de Lyon-Ecole Nationale Vétérinaire de Lyon (ENVL)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
viruses, lcsh:Medicine, MESH: Base Sequence, MESH: Reproduction, medicine.disease_cause, Polymerase Chain Reaction, law.invention, Dengue fever, Aedes, law, MESH : Insect Vectors, MESH : Female, MESH: Animals, MESH: In Situ Hybridization, Fluorescence, Chikungunya, lcsh:Science, MESH : Polymerase Chain Reaction, In Situ Hybridization, Fluorescence, Polymerase chain reaction, 0303 health sciences, Multidisciplinary, Reproduction, virus diseases, MESH : Reproduction, MESH: Aedes, Genitalia, Female, 3. Good health, MESH : In Situ Hybridization, Fluorescence, MESH : Genitalia, Female, MESH : Wolbachia, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Wolbachia, MESH : DNA Primers, Female, Wolbachia, Microbiology/Cellular Microbiology and Pathogenesis, Research Article, MESH: DNA Primers, Aedes albopictus, MESH: Insect Vectors, Biology, [ SDV.MP.VIR ] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Microbiology, 03 medical and health sciences, parasitic diseases, medicine, Animals, Microbiology/Environmental Microbiology, MESH : Aedes, DNA Primers, 030304 developmental biology, Base Sequence, 030306 microbiology, Host (biology), lcsh:R, MESH: Polymerase Chain Reaction, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, medicine.disease, Virology, Insect Vectors, Vector (epidemiology), lcsh:Q, MESH : Base Sequence, MESH : Animals, MESH: Genitalia, Female, MESH: Female, Bacteria
Abstract

International audience; BACKGROUND:Commensal and symbiotic microbes have a considerable impact on the behavior of many arthropod hosts, including hematophagous species that transmit pathogens causing infectious diseases to human and animals. Little is known about the bacteria associated with mosquitoes other than the vectorized pathogens. This study investigated Wolbachia and cultivable bacteria that persist through generations in Ae. albopictus organs known to host transmitted arboviruses, such as dengue and chikungunya.METHODOLOGY/PRINCIPAL FINDINGS:We used culturing, diagnostic and quantitative PCR, as well as in situ hybridization, to detect and locate bacteria in whole individual mosquitoes and in dissected tissues. Wolbachia, cultivable bacteria of the genera Acinetobacter, Comamonas, Delftia and Pseudomonas co-occurred and persisted in the bodies of both males and females of Ae. albopictus initially collected in La Réunion during the chikungunya outbreak, and maintained as colonies in insectaries. In dissected tissues, Wolbachia and the cultivable Acinetobacter can be detected in the salivary glands. The other bacteria are commonly found in the gut. Quantitative PCR estimates suggest that Wolbachia densities are highest in ovaries, lower than those of Acinetobacter in the gut, and approximately equal to those of Acinetobacter in the salivary glands. Hybridization using specific fluorescent probes successfully localized Wolbachia in all germ cells, including the oocytes, and in the salivary glands, whereas the Acinetobacter hybridizing signal was mostly located in the foregut and in the anterior midgut.CONCLUSIONS/SIGNIFICANCE:Our results show that Proteobacteria are distributed in the somatic and reproductive tissues of mosquito where transmissible pathogens reside and replicate. This location may portend the coexistence of symbionts and pathogens, and thus the possibility that competition or cooperation phenomena may occur in the mosquito vector Ae. albopictus. Improved understanding of the vectorial system, including the role of bacteria in the vector's biology and competence, could have major implications for understanding viral emergences and for disease control.

Published
2009
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