Descriptor: "MESH: Cytomegalovirus" - Searchworks@Jio Institute Digital Library Search Results

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32 results on '"MESH: Cytomegalovirus"'

1. Sémiologie et pronostic de l’infection congénitale à cytomégalovirus après primo-infection maternelle du second trimestre de la grossesse

Author: Cornet, Nathalie, Université Paris Descartes - Paris 5 (UPD5), and Yves Ville
Subjects: Diagnostic Imaging, MESH: Cytomegalovirus, Small For Gestational Age, Infection congénitale, MESH: Infant, small for gestational age, Congenital Infection, Pronostic, Cytomegalovirus, Petit poids pour l’âge gestationnel, MESH: Toxoplasmosis, Congenital, Imagerie anténatale, MESH: Prognosis, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: Congenital CMV is a major cause of neurological and sensory impairment in children. Objective - Description of the symptomatology and radiological semiology of infected fetuses in the second trimester of pregnancy; assessment of the prevalence and neonatal and long-term prognosis of fetuses with intrauterine growth restriction (IUGR) and small for gestational age birth weight (SGA). Material and Method - Monocentric retrospective study including patients referred for CMV seroconversion in the second trimester of pregnancy with a positive CMV viral load in amniotic fluid and a known neonatal outcome. Results - 111 fetuses had positive amniocentesis for CMV, 33 of them after second trimester seroconversion (29.7%). 21.2% of the fetuses were symptomatic at birth but none were long-term symptomatic. Radiological semiology, essentially composed of extracerebral signs, was a prognostic factor for birth symptomatology in univariate analysis. In utero, fetuses with IUGR were significantly more symptomatic at birth than eutrophic fetuses (64.7% vs. 12.1%, p
Published: 2019

2. Repertoire characterization and validation of gB-specific human IgGs directly cloned from humanized mice vaccinated with dendritic cells and protected against HCMV

Author: Torsten Witte, Martin Messerle, Andreas Schneider, Johannes Koenig, Sebastian J. Theobald, Michael Mach, Matthias Ballmaier, Simon Danisch, Henning Olbrich, Renata Stripecke, Agnes Bonifacius, Lutz Gieselmann, Constanca Figueiredo, Michael Meyer-Hermann, Meryem S. Ercanoglu, Constantin von Kaisenberg, Frank Klawonn, Britta Eiz-Vesper, Florian Klein, Marija Backovic, Christoph Kreer, Sahamoddin Khailaie, Valery Volk, Hannover Medical School [Hannover] (MHH), German Center for Infection Research - partner site Hannover-Braunschweig (DZIF), University of Cologne, University Hospital of Cologne [Cologne], Helmholtz Centre for Infection Research (HZI), Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Virologie Structurale - Structural Virology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), German Center for Infection Research - Partner Site Bonn-Cologne (DZIF), Ostfalia University of Applied Sciences, Technische Universität Braunschweig = Technical University of Braunschweig [Braunschweig], R.S./S.T./V.V./H.O. Hannover: This work was financed by grants of the German Center for Infections Research (DZIF-TTU07.803 and DZIF-TTU07.805 to R.S.), by a research collaboration grant of 'The Jackson Laboratory' and by the German Research Council (DFG/SFB738 Project A6 to R.S. and MM, DFG/REBIRTH Unit 6.4 to R.S.). S.T. received a RegSci Ph.D. fellowship, H.O. received a DZIF-Strucmed fellowship and V.V. received a DAAD/ZIB Ph.D. fellowship. F.K./C.K./ Univ. Cologne: This work was funded by grants from the German Center for Infection Research (DZIF, F.K.), the German Research Foundation (CRC 1279, F.K., CRC 1310, C.K. and F.K., Heisenberg-Program KL2389/2-1, F.K.) and the European Research Council (ERC-StG639961, F.K.). M.M.-H./ S.K./ Braunschweig: S.K. was supported by the German Federal Ministry of Education and Research (BMBF) for the eMED project SYSIMIT and by the Helmholtz-Gemeinschaft, Zukunftsthema 'Immunology and Inflammation' (ZT-0027)., European Project: 639961,H2020,ERC-2014-STG,HIV1ABTHERAPY(2016), BRICS, Braunschweiger Zentrum für Systembiologie, Rebenring 56,38106 Braunschweig, Germany, HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany., and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: B Cells, Pathology and Laboratory Medicine, Antibodies, Viral, Biochemistry, MESH: Antibodies, Monoclonal, MESH: Antibodies, Neutralizing, Mice, Animal Cells, Cytotoxic T cell, MESH: Animals, Biology (General), Enzyme-Linked Immunoassays, Immune Response, MESH: Immunoglobulin G, 0303 health sciences, 030302 biochemistry & molecular biology, Antibodies, Monoclonal, 3. Good health, Medical Microbiology, Viral Pathogens, MESH: Immunization, Passive, Antibody, Blood cells, QH301-705.5, T cell, Immunology, Cytotoxic T cells, Microbiology, 03 medical and health sciences, Antigen, Genetics, Humans, Antibody-Producing Cells, Immunoassays, Molecular Biology, Microbial Pathogens, MESH: Humans, Organisms, Immunization, Passive, Proteins, Correction, MESH: Cytomegalovirus Infections, Dendritic Cells, biochemical phenomena, metabolism, and nutrition, medicine.disease, Virology, Antibodies, Neutralizing, Animal Studies, Parasitology, Immunologic diseases. Allergy, MESH: Disease Models, Animal, MESH: Antibodies, Viral, Human cytomegalovirus, Physiology, [SDV]Life Sciences [q-bio], Cytomegalovirus, White Blood Cells, Cytomegalovirus Vaccines, Medizinische Fakultät, Immune Physiology, Cellular types, Medicine and Health Sciences, Antigens, Viral, Immune System Proteins, biology, MESH: Dendritic Cells, T Cells, Immune cells, Animal Models, medicine.anatomical_structure, Experimental Organism Systems, Viruses, Cytomegalovirus Infections, Human Cytomegalovirus, Pathogens, MESH: Antigens, Viral, Research Article, MESH: Cytomegalovirus, Cell biology, Herpesviruses, Somatic hypermutation, Mouse Models, Research and Analysis Methods, Antibodies, Immune system, Model Organisms, MESH: Cytomegalovirus Vaccines, medicine, Animals, ddc:610, MESH: Mice, 030304 developmental biology, Biology and life sciences, RC581-607, Disease Models, Animal, Immunization, Immunoglobulin G, biology.protein, Immunologic Techniques, DNA viruses
Abstract: Human cytomegalovirus (HCMV) causes serious complications to immune compromised hosts. Dendritic cells (iDCgB) expressing granulocyte-macrophage colony-stimulating factor, interferon-alpha and HCMV-gB were developed to promote de novo antiviral adaptive responses. Mice reconstituted with a human immune system (HIS) were immunized with iDCgB and challenged with HCMV, resulting into 93% protection. Immunization stimulated the expansion of functional effector memory CD8+ and CD4+ T cells recognizing gB. Machine learning analyses confirmed bone marrow T/CD4+, liver B/IgA+ and spleen B/IgG+ cells as predictive biomarkers of immunization (≈87% accuracy). CD8+ and CD4+ T cell responses against gB were validated. Splenic gB-binding IgM-/IgG+ B cells were sorted and analyzed at a single cell level. iDCgB immunizations elicited human-like IgG responses with a broad usage of various IgG heavy chain V gene segments harboring variable levels of somatic hypermutation. From this search, two gB-binding human monoclonal IgGs were generated that neutralized HCMV infection in vitro. Passive immunization with these antibodies provided proof-of-concept evidence of protection against HCMV infection. This HIS/HCMV in vivo model system supported the validation of novel active and passive immune therapies for future clinical translation., Author summary Human cytomegalovirus (HCMV) is a ubiquitous pathogen. As long as the immune system is functional, T and B cells can control HCMV. Yet, for patients who have debilitated immune functions, HCMV infections and reactivations cause major complications. Vaccines or antibodies to prevent or treat HCMV are not yet approved. Novel animal models for testing new immunization approaches are emerging and are important tools to identify biomedical products with a reasonable chance to work in patients. Here, we used a model based on mice transplanted with human immune cells and infected with a traceable HCMV. We tested a cell vaccine (iDCgB) carrying gB, a potent HCMV antigen. The model showed that iDCgB halted the HCMV infection in more than 90% of the mice. We found that antibodies were key players mediating protection. Using state-of-the-art methods, we were able to use the sequences of the human antibodies generated in the mice to construct and produce monoclonal antibodies in the laboratory. Proof-of-concept experiments indicated that administration of these monoclonal antibodies into mice protected them against HCMV infection. In summary, this humanized mouse model was useful to test a vaccine and to generate and test novel antibodies that can be further developed for human use.
Published: 2019
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3. Pronostic du fœtus infecté par le cytomégalovirus après primo-infection maternelle du premier trimestre

Author: Faure Bardon, Valentine, Université Paris Descartes - Faculté de Médecine (UPD5 Médecine), Université Paris Descartes - Paris 5 (UPD5), and Marianne Leruez-Ville
Subjects: MESH: Cytomegalovirus, Congénital, Pronostic, MESH: Congenital, Hereditary, and Neonatal Diseases and Abnormalities, Cytomégalovirus, MESH: Prognosis, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: L’infection congénitale à cytomégalovirus (CMV) peut entrainer des séquelles chez l’enfant tels qu’un déficit auditif et un retard mental. En France, la prévalence chez le nouveau-né est de 0,37 % et fait suite dans 52 % des cas à une primo-infection maternelle (PIM) et dans 48 % à une infection maternelle secondaire (IMS). L’objectif de ce travail était de définir, grâce aux moyens disponibles en anténatal, le pronostic du fœtus infecté par le CMV après PIM. Dans une première partie nous avons évalué le trimestre de contamination comme facteur pronostique. Nous avons conduit une étude sur 255 patientes et leurs 260 fœtus / 234 nouveau-nés infectés et nous avons montré qu’après PIM, seuls les cas d’infection du 1er trimestre ont présenté des séquelles auditives et/ou neurologiques avec un risque de 32,4 %. Dans une seconde partie, nous avons cherché à préciser le risque de séquelles dans cette population à haut risque en fonction des résultats des examens disponibles en prénatal : échographies, IRM, biologie fœtale. Dans une étude rétrospective, longitudinale, standardisée de 62 fœtus infectés après PIM du 1er trimestre, nous avons montré qu’un suivi anténatal sans aucune anomalie décelable permettait de donner un conseil prénatal rassurant défini comme l’absence de séquelle modérée à sévère. Nous avons montré que les séquelles, après PIM, n’étaient présentes que chez les enfants infectés lors du 1er trimestre et que le pronostic de ces fœtus pouvait être affiné grâce à une combinaison d’examens disponibles en prénatal. Des études s’intéressant spécifiquement aux IMS sont nécessaires afin de compléter ce travail d’évaluation pronostique de l’infection congénitale à CMV.
Published: 2019

4. CMV et grossesse : connaissances et pratiques des professionnels de santé

Author: Fellah, Thanina, Université Paris Descartes - École de sages-femmes Baudelocque (UPD ESF Baudelocque), Université Paris Descartes - Paris 5 (UPD5), Université Paris Descartes - Faculté de Médecine (UPD5 Médecine), and Olivier Picone
Subjects: MESH: Cytomegalovirus, Grossesse, Prévention, MESH: Pregnancy, Congenital infection, Infection congénitale, Prevention, Professionnels de santé, Connaissances, CMV, MESH: Knowledge, [SDV.MHEP.GEO]Life Sciences [q-bio]/Human health and pathology/Gynecology and obstetrics, Health care providers
Abstract: Objectives: the main objective of our work was to evaluate the knowledge and practices of health care providers regarding CMV during pregnancy. Our secondary objective was to compare the results obtained with those of the Cordier et al. study (1) Material and method: quantitative, cross-sectional, observational and multicentric study conducted through an online questionnaire. In order to quantify the knowledge of the interviewees we have set up a 30-point score. Results: we received 597 responses including 199 obstetricians, 17 medical gynecologists, 305 midwives, 29 pediatricians, 7 biologists, 32 interns. Out of the total population (n=597), we found 82.4% who obtained a score greater than or equal to 15/30 on the score used to assess knowledged By including only the gynecologists, obstetricians and midwives, it was found that people with a score superior or equal to 15/30 were informing more the patients about CMV than professionals with less than 15/30 (p=0.005). Concerning practices, 353/521 professionals working in perinatal care provide information about CMV to patients. Professionals working in private practice provide more often information than those working in hospitals (p
Published: 2019

5. Conserved domains and structure prediction of human cytomegalovirus UL27 protein

Author: Gaël Champier, Anthony Couvreux, Laurène Trapes, Sébastien Cotin, Sébastien Hantz, François Denis, Sophie Alain, Serge Bouaziz, Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, Laboratoire d'informatique Fondamentale de Marseille - UMR 6166 (LIF), Université de la Méditerranée - Aix-Marseille 2-Université de Provence - Aix-Marseille 1-Centre National de la Recherche Scientifique (CNRS), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-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), 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)-Institut de Chimie du CNRS (INC), Bouaziz, Serge, Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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), and 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)-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)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Human cytomegalovirus, MESH: Sequence Analysis, DNA, viruses, Cytomegalovirus, MESH: Amino Acid Sequence, medicine.disease_cause, chemistry.chemical_compound, MESH: Structure-Activity Relationship, Pharmacology (medical), Peptide sequence, Conserved Sequence, Mutation, MESH: Conserved Sequence, MESH: Drug Resistance, Viral, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], virus diseases, Maribavir, Cell biology, Phosphotransferases (Alcohol Group Acceptor), Infectious Diseases, MESH: Antiviral Agents, MESH: Cytomegalovirus, MESH: Mutation, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Sequence analysis, Molecular Sequence Data, MESH: Sequence Alignment, Sequence alignment, Biology, Antiviral Agents, Herpesviridae, Structure-Activity Relationship, Drug Resistance, Viral, medicine, Humans, Amino Acid Sequence, Pharmacology, MESH: Ribonucleosides, MESH: Humans, MESH: Molecular Sequence Data, Sequence Analysis, DNA, medicine.disease, Virology, MESH: Phosphotransferases (Alcohol Group Acceptor), MESH: DNA, Viral, chemistry, DNA, Viral, Benzimidazoles, Ribonucleosides, MESH: Benzimidazoles, Sequence Alignment, DNA
Abstract: Background The human cytomegalovirus (HCMV) nuclear UL27 protein (pUL27) could be involved at the stage of nuclear egress. Maribavir is a new anti-HCMV drug that targets nuclear egress through direct inhibition of the HCMV serine–threonine kinase, UL97 protein (pUL97). Because maribavir- resistance-related mutations are observed in both proteins, pUL27 is thought to interfere with pUL97 activity; however, its mechanism of action remains unclear. Methods As there is no available crystal structure for pUL27 or any known structures of its homologous proteins, we attempted to identify pUL27 functional domains by sequence analysis, identification of conserved domains, structure prediction and matching with previously known maribavir resistance mutations. Results The UL27 sequence analysis of 20 HCMV wild-type strains and 8 ganciclovir-resistant HCMV strains allowed us to describe four conserved domains, to localize the putative phosphorylation sites and to identify protein–protein interface domains, suggesting that pUL27 could interact with either pUL97 or itself. Conclusions Although the function of pUL27 is still unknown in the HCMV replication cycle, our approach identified target domains that appeared to be essential to the function of pUL27. This work provides a better understanding on the relative importance of each pUL27 mutation and could form the basis of later comparison analyses, when a three-dimensional structure of a pUL27 homologue will be available.
Published: 2019

6. Étude des facteurs prédictifs de séquelles à 2 ans en lien avec une infection congénitale à Cytomégalovirus

Author: Petermann, Lucile, Université Paris Descartes - Faculté de Médecine (UPD5 Médecine), Université Paris Descartes - Paris 5 (UPD5), and Jean-François Magny
Subjects: MESH: Cytomegalovirus, MESH: Immunologic Deficiency Syndromes, Dépistage sérologique systématique, MESH: Maternal Serum Screening Tests, Handicap, Symptomatic, Trouble neuro-développemental, MESH: Prognosis, Hearing impairment, Congenital infection, Infection congénitale, MESH: Risk Factors, Pronostic, MESH: Therapeutics, Traitement, MESH: Hearing Loss, MESH: Neurodevelopmental Disorders, Facteurs de risques, MESH: Cytomegalovirus Infections, MESH: Disabled Persons, Cytomégalovirus, Déficience, Déficience auditive, Impairment, Symptomatique, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: Objectives: cytomegalovirus (CMV) is the leading cause of congenital infection in the world with a major risk of sequelae including neurological and auditory. The aim of the study is to identify what are the prognostic factors for defining neonates at risk of developing sequelae at 2 years related to congenital CMV infection. Design of the study: monocentric retrospective observational study including 220 patients born between January 2001 and October 2015 with congenital CMV infection. We studied various pre- and postnatal factors and their impact on the outcome at 2 years of age for children with congenital CMV infection. Results: the presence of at least one postnatal abnormality (clinical examination or additional investigations) was a risk factor for developing sequelae at 2 years (p = 0.037). Neonates with maternal seroconversion in the first trimester were more likely to develop sequelae at 2 years (p
Published: 2018

7. Aminobisphosphonates Synergize with Human Cytomegalovirus To Activate the Antiviral Activity of Vγ9Vδ2 Cells

Author: Suzanne Peyrottes, Eric Champagne, Charline Daguzan, Christian Davrinche, Morgane Moulin, Hanna Kulyk-Barbier, Centre de Physiopathologie Toulouse Purpan (ex IFR 30 et IFR 150) (CPTP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre de Physiopathologie Toulouse Purpan ex IFR 30 et IFR 150 (CPTP), Institut National de la Santé et de la Recherche Médicale (INSERM), Université Montpellier 2 - Sciences et Techniques (UM2), Ecole Nationale Supérieure de Chimie de Montpellier, and Grant Number : Fondation pour la Recherche Medicale Grant [DCM20121225761], Comité d'éthique de l'Inserm (CEI), 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), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM), 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), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), and Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)
Subjects: Cytotoxicity, Immunologic, 0301 basic medicine, Human cytomegalovirus, lymphocytes, bisphosphonate, [SDV]Life Sciences [q-bio], Isopentenyl pyrophosphate, Cytomegalovirus, hiv-infection, MESH: T-Lymphocyte Subsets, Lymphocyte Activation, Virus Replication, stimulation, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, chemistry.chemical_compound, zoledronic acid, T-Lymphocyte Subsets, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Immunology and Allergy, Cells, Cultured, ComputingMilieux_MISCELLANEOUS, MESH: Cytokines, Diphosphonates, MESH: Dendritic Cells, Receptors, Antigen, T-Cell, gamma-delta, tumor-cells, MESH: Mevalonic Acid, inhibition, 3. Good health, MESH: Leukocytes, Mononuclear, medicine.anatomical_structure, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Cytomegalovirus Infections, Host-Pathogen Interactions, Cytokines, [SDV.IMM]Life Sciences [q-bio]/Immunology, Tumor necrosis factor alpha, receptor-gamma, delta t-cells, MESH: Cells, Cultured, MESH: Cytomegalovirus, replication, MESH: Diphosphonates, T cell, Immunology, Mevalonic Acid, Biology, Cell Line, 03 medical and health sciences, Antigen, MESH: Receptors, Antigen, T-Cell, gamma-delta, medicine, Humans, MESH: Cytotoxicity, Immunologic, MESH: Lymphocyte Activation, MESH: Humans, T-cell receptor, MESH: Virus Replication, MESH: Host-Pathogen Interactions, Lymphokine, Dendritic Cells, MESH: Cytomegalovirus Infections, [SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy, medicine.disease, Virology, MESH: Cell Line, 030104 developmental biology, chemistry, Cell culture, Leukocytes, Mononuclear, Cancer research, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract: Human V gamma 9V delta 2 T cells are activated through their TCR by neighboring cells producing phosphoantigens. Zoledronate (ZOL) treatment induces intracellular accumulation of the phosphoantigens isopentenyl pyrophosphate and ApppI. Few attempts have been made to use immunomanipulation of V gamma 9V delta 2 lymphocytes in chronic viral infections. Although V gamma 9V delta 2 T cells seem to ignore human CMV (HCMV)-infected cells, we examined whether they can sense HCMV when a TCR stimulus is provided with ZOL. Fibroblasts treated with ZOL activate V gamma 9V delta 2 T cells to produce IFN-gamma but not TNF. Following the same treatment, HCMV-infected fibroblasts stimulate TNF secretion and an increased production of IFN-g, indicating that V gamma 9V delta 2 cells can sense HCMV infection. Increased lymphokine production was observed with most clinical isolates and laboratory HCMV strains, HCMV-permissive astrocytoma, or dendritic cells, as well as "naive" and activated V gamma 9V delta 2 cells. Quantification of intracellular isopentenyl pyrophosphate/ApppI following ZOL treatment showed that HCMV infection boosts their accumulation. This was explained by an increased capture of ZOL and by upregulation of HMG-CoA synthase and reductase transcription. Using an experimental setting where infected fibroblasts were cocultured with gamma delta cells in submicromolar concentrations of ZOL, we show that V gamma 9V delta 2 cells suppressed substantially the release of infectious particles while preserving uninfected cells. V gamma 9V delta 2 cytotoxicity was decreased by HCMV infection of targets whereas anti-IFN-gamma and anti-TNF Abs significantly blocked the antiviral effect. Our experiments indicate that cytokines produced by V gamma 9V delta 2 T cells have an antiviral potential in HCMVinfection. This should lead to in vivo studies to explore the possible antiviral effect of immunostimulation with ZOL in this context.
Published: 2016
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8. Therapeutic drug monitoring of ganciclovir in the management of cytomegalovirus : should we reassess the therapeutic range ? Results of a preliminary study conducted at the University Hospital of Rennes

Author: Arnoult, Claire, Université de Rennes 1 - Faculté des sciences pharmaceutiques et biologiques (UR1 Pharmacie), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), and Florian Lemaitre
Subjects: MESH: Relation dose-effet des médicaments, MESH: Cytomegalovirus, Médicaments -- Relations dose-effet, Cytomégalovirus humain, [SDV]Life Sciences [q-bio], MESH: Ganciclovir, virus diseases, Pharmacologie, Ganciclovir, MESH: Surveillance pharmacologique
Abstract: Cytomegalovirus (CMV) is responsible for potentially severe infections in immunocompromised patients including transplant recipients and AIDS patients. Ganciclovir represents the first-line treatment in the management of these infections. However, the increasing number of resistances highlights the need to use new molecules and find new strategies more effective. Therapeutic drug monitoring is used to obtain an optimal patient exposure to ganciclovir. This work aims to assess the impact of greater ganciclovir exposure on the effectiveness of treatment. The retrospective study compares the evolution of CMV viral load between patients exposed to normal trough plasma concentrations and patients exposed to higher concentrations.; Le cytomegalovirus (CMV) est responsable d'infections potentiellement graves chez les personnes immunodéprimées notamment chez les patients greffés ou atteints du SIDA. Le ganciclovir représente la première ligne de traitement dans la prise en charge de ces infections. Cependant, la mise en évidence de résistances entraine l'utilisation de nouvelles molécules et la recherche de nouvelles stratégies plus efficaces. Le suivi thérapeutique pharmacologique permet d'obtenir une exposition optimale du patient au ganciclovir et représente ainsi une stratégie utile dans la prise en charge du cytomegalovirus. Ce travail a pour objectif d'évaluer si une exposition au ganciclovir plus importante que celle habituellement utilisée n'améliorerait pas l'efficacité du traitement. L'étude réalisée compare l'évolution de la charge virale plasmatique CMV entre des patients exposés à des concentrations plasmatiques résiduelles habituelles et des patients exposés à des concentrations plus élevées.
Published: 2016

9. Development and validation of a non-radioactive DNA polymerase assay for studying cytomegalovirus resistance to foscarnet

Author: Alexandra Ducancelle, Marie-Christine Mazeron, Marie-José Sanson-Le Pors, Françoise Petit, Sophie Alain, Hôpital Lariboisière, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Lariboisière-Université Paris Diderot - Paris 7 (UPD7), Université de Limoges (UNILIM), Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Lariboisière-Fernand-Widal [APHP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)
Subjects: Foscarnet, Human cytomegalovirus, DNA polymerase, Cytomegalovirus, MESH: Catalytic Domain, DNA-Directed DNA Polymerase, chemistry.chemical_compound, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Catalytic Domain, MESH: Colorimetry, MESH: DNA-Directed DNA Polymerase, Polymerase, MESH: Inhibitory Concentration 50, 0303 health sciences, MESH: Drug Resistance, Viral, biology, virus diseases, 3. Good health, MESH: Reproducibility of Results, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Colorimetry, MESH: Foscarnet, medicine.drug, MESH: Antiviral Agents, MESH: Cytomegalovirus, MESH: Mutation, Antiviral Agents, Sensitivity and Specificity, Inhibitory Concentration 50, Viral Proteins, 03 medical and health sciences, Virology, Drug Resistance, Viral, medicine, Gene, 030304 developmental biology, 030306 microbiology, MESH: Digoxigenin, Mutagenesis, Reproducibility of Results, biochemical phenomena, metabolism, and nutrition, medicine.disease, MESH: Viral Proteins, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Molecular biology, MESH: Sensitivity and Specificity, chemistry, Foscarnet Sodium, Mutation, biology.protein, Digoxigenin, DNA
Abstract: International audience; Phenotypic characterisation of the human cytomegalovirus (HCMV) pUL54 DNA polymerase is a useful tool for testing for mutations in the UL54 gene thought to render HCMV resistant to foscarnet. In this study, an in-house non-isotopic method for assessing polymerase enzymatic activity in the presence and absence of foscarnet was developed and its utility for HCMV polymerase phenotyping evaluated. Polymerase activity was assessed by monitoring the incorporation of digoxigenin-labelled nucleotides into the growing DNA chain and foscarnet concentrations inhibiting enzymatic activity by 50% were determined. HCMV DNA polymerases were synthesised in vitro by expression of UL54 under the control of the T7 promoter. Mutations of interest were introduced into the wild-type UL54 gene by site-directed mutagenesis. Mutated polymerases and polymerases from HCMV reference strains were studied. The activity of polymerases containing mutations known to confer resistance to foscarnet (V715M, T700A and N495K) was inhibited by concentrations of foscarnet eight to 14 times higher than those required to inhibit wild-type polymerases. Our in-house non-radioactive phenotypic assay was sensitive and reproducible. It is also easy to perform and could provide a convenient method for characterising mutations conferring resistance to foscarnet in HCMV.
Published: 2007
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10. Retrograde transport is not required for cytosolic translocation of the B-subunit of Shiga toxin

Author: Estelle Dransart, Eric Tartour, Henri-François Renard, Stefan J. Rathjen, Bahne Stechmann, Thi Tran, Mike Lord, Ludger Johannes, Jean-Christophe Cintrat, Jost Enninga, Alexandre Bobard, Maria Daniela Garcia-Castillo, Christophe Lamaze, Chimie biologique des membranes et ciblage thérapeutique (CBMCT - UMR 3666 / U1143), Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Institut Curie [Paris], Paris-Centre de Recherche Cardiovasculaire (PARCC - UMR-S U970), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Université Paris Descartes - Paris 5 (UPD5), Université Sorbonne Paris Cité (USPC), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Dynamique des Interactions Hôte-Pathogène - Dynamics of Host-Pathogen Interactions, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), University of Warwick [Coventry], Service de Chimie Bio-Organique et de Marquage (SCBM), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-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 Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-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 Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), This work was supported by grants from the Institut National du Cancer [grant number PLBIO11-022-IDF-JOHANNES], Agence Nationale pour la Recherche [grant number ANR-11 BSV2 018 03], European Research Council (ERC) advanced grant (project 340485) to L.J., and by fellowships from AXA Research Funds and Association pour la Recherche sur le Cancer to M.D.G-C. The Johannes team is members of Labex CelTisPhyBio [grant number 11-LBX-0038] and of Idex Paris Sciences et Lettres [grant number ANR-10-IDEX-0001-02 PSL]. The Tartour team is members of Labex Immuno-Oncology and SIRIC-CARPEM and is labeled by the Ligue Nationale contre le Cancer. J.E. is supported by an ERC starting grant [Rupteffect, grant number 261166] and is member of the LabEx initiative IBEID. His group has been supported by the CARNOT-MIE program., We also thank the following people for their help with experiments and reagents, as well as scientific discussion: Bruno Goud, Stéphanie Miserey-Lenkei, Sebastian Amigorena and Daniel Gillet., ANR-11-IDEX-0005,USPC,Université Sorbonne Paris Cité(2011), ANR-10-IDEX-0001,PSL,Paris Sciences et Lettres(2010), ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), European Project: 340485,EC:FP7:ERC,ERC-2013-ADG,GALECTCOMPART(2014), European Project: 261166,EC:FP7:ERC,ERC-2010-StG_20091118,RUPTEFFECTS(2011), Langlais, Laurence, Université Sorbonne Paris Cité - - USPC2011 - ANR-11-IDEX-0005 - IDEX - VALID, Initiative d'excellence - Paris Sciences et Lettres - - PSL2010 - ANR-10-IDEX-0001 - IDEX - VALID, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, Endocytic Membrane Compartmentalization by Galectins - GALECTCOMPART - - EC:FP7:ERC2014-04-01 - 2019-03-31 - 340485 - VALID, Revealing the mechanism of host membrane rupture by invasive pathogens and its role in triggering the immune response - RUPTEFFECTS - - EC:FP7:ERC2011-03-01 - 2016-02-29 - 261166 - VALID, Université Paris Descartes - Paris 5 (UPD5)-Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and UCL - SST/ISV - Institut des sciences de la vie
Subjects: Lactamase, MESH: Fluorescence Resonance Energy Transfer, Retro compound, Cytomegalovirus, Chromosomal translocation, CD8-Positive T-Lymphocytes, [SDV.BC.IC] Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Sec22B, Epitopes, chemistry.chemical_compound, MESH: Cell Compartmentation, Cytosol, 0302 clinical medicine, Rab5, MESH: Cytosol, Rab6, Rab7, Cytotoxic T-lymphocyte, [SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB], Fluorescence Resonance Energy Transfer, Golgi, Endosome, PPMP, Microdomain, [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Cancer, 0303 health sciences, Infectious disease, biology, Brefeldin A, Endosomal escape, MESH: CD8-Positive T-Lymphocytes, Endocytosis, Cell biology, Methyl-beta-cyclodextrin, Cholesterol, Biochemistry, Nanodomain, MESH: Protein Biosynthesis, MESH: Endocytosis, Ribosome Inactivating Proteins, Type 1, symbols, Immunotherapy, Bafilomycin A1, Dendritic cell, MESH: Cytomegalovirus, MESH: Epitopes, MESH: Biological Transport, MESH: Shiga Toxin, Endosomes, MESH: Ribosome Inactivating Proteins, Type 1, Major histocompatibility complex, Shiga Toxin, 03 medical and health sciences, symbols.namesake, Antigen, Humans, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Raft, 030304 developmental biology, MESH: Humans, Endoplasmic reticulum, rab7 GTP-Binding Proteins, Biological Transport, Cell Biology, Golgi apparatus, Shiga toxin, Saporins, Cell Compartmentation, MESH: rab GTP-Binding Proteins, Antigen cross-presentation, chemistry, rab GTP-Binding Proteins, MESH: Endosomes, Protein Biosynthesis, MESH: HeLa Cells, biology.protein, Brefeldin-A, 030217 neurology & neurosurgery, HeLa Cells
Abstract: Antigen-presenting cells have the remarkable capacity to transfer exogenous antigens to the cytosol for processing by proteasomes and subsequent presentation on major histocompatibility complex class-I (MHC-I) molecules, a process termed cross-presentation. This is the target of biomedical approaches that aim to trigger a therapeutic immune response. The receptor-binding B-subunit of Shiga toxin (STxB) has been developed as an antigen delivery tool for such immunotherapy applications. In this study, we have analyzed pathways and trafficking factors that are involved in this process. A covalent conjugate between STxB and saporin was generated to quantitatively sample the membrane translocation step to the cytosol in differentiated monocyte-derived THP-1 cells. We have found that retrograde trafficking to the Golgi complex was not required for STxB–saporin translocation to the cytosol or for STxB-dependent antigen cross-presentation. Depletion of endosomal Rab7 inhibited, and lowering membrane cholesterol levels favored STxB–saporin translocation. Interestingly, experiments with reducible and non-reducible linker-arm–STxB conjugates led to the conclusion that after translocation, STxB remains associated with the cytosolic membrane leaflet. In summary, we report new facets of the endosomal escape process bearing relevance to antigen cross-presentation.
Published: 2015
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11. A Novel Mutation in the Ul54 Gene of Human Cytomegalovirus Isolates that Confers Resistance to Foscarnet

Author: Marie-José Sanson-Le Pors, Françoise Petit, Sophie Alain, Gaël Champier, Marie-Christine Mazeron, Alexandra Ducancelle, Bucek, Ingrid, Hôpital Lariboisière, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Université de Limoges (UNILIM), Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Lariboisière-Université Paris Diderot - Paris 7 (UPD7)
Subjects: Foscarnet, Human cytomegalovirus, viruses, Cytomegalovirus, DNA-Directed DNA Polymerase, Drug resistance, Recombinant virus, chemistry.chemical_compound, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: DNA-Directed DNA Polymerase, Pharmacology (medical), Recombination, Genetic, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Microbial Sensitivity Tests, MESH: Drug Resistance, Viral, virus diseases, Infectious Diseases, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Cytomegalovirus Infections, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Recombination, Genetic, MESH: Foscarnet, Cidofovir, medicine.drug, MESH: Antiviral Agents, Ganciclovir, MESH: Cytomegalovirus, MESH: Mutation, Microbial Sensitivity Tests, Biology, Antiviral Agents, Virus, Viral Proteins, Drug Resistance, Viral, medicine, Humans, Pharmacology, MESH: Humans, MESH: Cytomegalovirus Infections, Fibroblasts, biochemical phenomena, metabolism, and nutrition, medicine.disease, MESH: Viral Proteins, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Virology, chemistry, MESH: Fibroblasts, Foscarnet Sodium, Mutation, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
Abstract: International audience; Foscarnet is currently licensed for the treatment of human cytomegalovirus (HCMV) infection. Mutations proven to confer resistance to foscarnet have mostly been mapped to regions II, III and VI of the HCMV UL54-encoded DNA polymerase. We previously showed that sequential foscarnet-resistant HCMV isolates recovered from a patient with lymphoma had change N495K in region delta-C of the DNA polymerase. To evaluate the impact of change N495K on HCMV sensitivity to foscarnet, a recombinant HCMV strain carrying the mutation was produced by homologous recombination. The recombinant virus showed a 3.4-fold increase in foscarnet resistance, and remained sensitive to ganciclovir and cidofovir. In addition, the recombinant strain showed a reduction of infectious virus yield compared with its parent strain. Change N495K should be added to the list of mutations conferring resistance to foscarnet and be taken into account in the genotypic diagnosis of antiviral resistance.
Published: 2005
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12. Human Cytomegalovirus Binding to DC-SIGN Is Required for Dendritic Cell Infection and Target Cell trans-Infection

Author: Thierry Delaunay, Franck Fieschi, Julie Déchanet-Merville, Fernando Arenzana-Seisdedos, Ali Amara, Franck Halary, Martin Messerle, Corinne Houles, Jean Francois Moreau, Hugues Lortat-Jacob, Génomique, développement et pouvoir pathogène (GD2P), Université Bordeaux Segalen - Bordeaux 2-Institut National de la Recherche Agronomique (INRA), Composantes innées de la réponse immunitaire et différenciation (CIRID), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075), 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)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie et Biochimie Moléculaires et Supramoléculaires (ICBMS), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), Pathogénie Virale, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Pathologies infectieuses et cancers : aspects biologiques et thérapeutiques (PICABT), Université Bordeaux Segalen - Bordeaux 2-CHU Bordeaux [Bordeaux]-Institut Bergonié - CRLCC Bordeaux-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Hétérochimie moléculaire et macromoléculaire (HMM), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), 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)-Centre National de la Recherche Scientifique (CNRS), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-École Supérieure Chimie Physique Électronique de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université Bordeaux Segalen - Bordeaux 2-CHU Bordeaux [Bordeaux]-Institut Bergonié [Bordeaux], UNICANCER-UNICANCER-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-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), Université de Lyon-Université de Lyon-École Supérieure de Chimie Physique Électronique de Lyon (CPE)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Pathogénie Virale - Viral Pathogenesis, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Bergonié [Bordeaux], and UNICANCER-UNICANCER-Université Bordeaux Segalen - Bordeaux 2-CHU Bordeaux [Bordeaux]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Human cytomegalovirus, [SDV]Life Sciences [q-bio], Cytomegalovirus, Ligands, Virus Replication, IMMUNOLOGIE, 0302 clinical medicine, Viral Envelope Proteins, MESH: Ligands, Immunology and Allergy, ComputingMilieux_MISCELLANEOUS, MESH: Receptors, Cell Surface, 0303 health sciences, biology, MESH: Dendritic Cells, Cell adhesion molecule, CMV, virus diseases, 3. Good health, DC-SIGN, Infectious Diseases, Cytomegalovirus Infections, MESH: Cell Adhesion Molecules, MESH: Cytomegalovirus, Immunology, Congenital cytomegalovirus infection, Receptors, Cell Surface, Virus, 03 medical and health sciences, Immune system, medicine, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Lectins, C-Type, 030304 developmental biology, MESH: Humans, MESH: Virus Replication, Dendritic cell, MESH: Cytomegalovirus Infections, Dendritic Cells, medicine.disease, Virology, Viral replication, MESH: Viral Envelope Proteins, MESH: HeLa Cells, biology.protein, Cell Adhesion Molecules, MESH: Lectins, C-Type, 030215 immunology, HeLa Cells
Abstract: International audience; Cytomegalovirus (CMV) infection is characterized by host immunosuppression and multiorganic involvement. CMV-infected dendritic cells (DC) were recently shown to display reduced immune functions, but their role in virus dissemination is not clear. In this report, we demonstrated that CMV could be captured by DC through binding on DC-SIGN and subsequently transmitted to permissive cells. Moreover, blocking DC-SIGN by specific antibodies inhibited DC infection by primary CMV isolates and expression of DC-SIGN or its homolog DC-SIGNR rendered susceptible cells permissive to CMV infection. We demonstrated that CMV envelope glycoprotein B is a viral ligand for DC-SIGN and DC-SIGNR. These results provide new insights into the molecular interactions contributing to cell infection by CMV and extend DC-SIGN implication in virus propagation.
Published: 2002
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13. Success and failure of artesunate treatment in five transplant recipients with disease caused by drug-resistant cytomegalovirus

Author: Patrice Morand, Raphaëlle Germi, Paolo Malvezzi, A. Thiebaut, O. Epaulard, Julien Lupo, Sophie Alain, C. Mariette, C. Saint Raymond, J. P. Brion, Unit for Virus Host-Cell Interactions [Grenoble] (UVHCI), Centre National de la Recherche Scientifique (CNRS)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Université Joseph Fourier - Grenoble 1 (UJF), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U1172 Inserm - U837 (JPArc), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Lille Nord de France (COMUE)-Université de Lille, Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, Molécules bioactives, conception, isolement et synthèse (MBCIS), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Service des Maladies Infectieuses, CHU Grenoble, Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche Jean-Pierre AUBERT Neurosciences et Cancer - U837 (JPArc), and Université Lille Nord de France (COMUE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille
Subjects: Male, Foscarnet, Artesunate, Cytomegalovirus, Drug resistance, chemistry.chemical_compound, MESH: Treatment Outcome, 0303 health sciences, MESH: Middle Aged, MESH: Drug Resistance, Viral, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Hematopoietic Stem Cell Transplantation, virus diseases, MESH: Transplantation, Middle Aged, Viral Load, Artemisinins, 3. Good health, Treatment Outcome, Cytomegalovirus Infections, Female, MESH: Viral Load, medicine.drug, Cidofovir, Ganciclovir, MESH: Antiviral Agents, MESH: Cytomegalovirus, Congenital cytomegalovirus infection, MESH: Organ Transplantation, Antiviral Agents, 03 medical and health sciences, Virology, Drug Resistance, Viral, MESH: Artemisinins, medicine, Humans, MESH: Hematopoietic Stem Cell Transplantation, 030304 developmental biology, Pharmacology, Transplantation, MESH: Humans, 030306 microbiology, business.industry, Organ Transplantation, MESH: Cytomegalovirus Infections, medicine.disease, MESH: Male, Multiple drug resistance, chemistry, Immunology, business, MESH: Female
Abstract: International audience; Cytomegalovirus (CMV) strains resistant to ganciclovir, cidofovir and/or foscarnet were genotypically and phenotypically characterised in two haematopoietic stem cell transplant recipients and three solid-organ transplant recipients with CMV disease. The anti-malaria drug artesunate led to a favourable virological and clinical response in three cases with mild CMV diseases (fever and neutropaenia) but was ineffective in two fatal CMV diseases with lung involvement in spite of a decrease in the CMV DNA load in blood and bronchoalveolar fluid.
Published: 2014

14. Human herpes virus co-infection is associated with mortality in HIV-negative patients with Pneumocystis jirovecii pneumonia

Author: Pierre Fillatre, H. Leroy, Y. Le Tulzo, Stéphane Jouneau, Pierre Tattevin, Matthieu Revest, Florence Robert-Gangneux, Sylviane Chevrier, Sophie Minjolle, Arnaud Gacouin, Service des maladies infectieuses et réanimation médicale [Rennes] = Infectious Disease and Intensive Care [Rennes], CHU Pontchaillou [Rennes], Service de Parasitologie-Mycologie [Rennes], Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Hôpital Pontchaillou-CHU Pontchaillou [Rennes], Unité de maladies infectieuses, Centre d'Investigation Clinique [Rennes] (CIC), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale (INSERM)-Service des maladies infectieuses et réanimation médicale [Rennes] = Infectious Disease and Intensive Care [Rennes], CHU Pontchaillou [Rennes]-CHU Pontchaillou [Rennes], Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale (INSERM), Contaminants Chimiques, immunité et Inflammation, Institut de recherche en santé, environnement et travail (Irset), Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Université d'Angers (UA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Laboratoire de Virologie [Rennes] = Virology [Rennes], Fonction structure et inactivation d'ARN bactérien, CHU Pontchaillou [Rennes]-CHU Pontchaillou [Rennes]-Fonction, structure et inactivation d'ARN bactériens, Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université de Rennes (UR)-Hôpital Pontchaillou-CHU Pontchaillou [Rennes], Université de Rennes (UR)-Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rennes (UR)-Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale (INSERM)-Service des maladies infectieuses et réanimation médicale [Rennes] = Infectious Disease and Intensive Care [Rennes], Université de Rennes (UR)-Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale (INSERM), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université d'Angers (UA)-Université de Rennes (UR)-École des Hautes Études en Santé Publique [EHESP] (EHESP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and Le Corre, Morgane
Subjects: Male, ARDS, Cytomegalovirus, MESH: Herpes Simplex, law.invention, Cohort Studies, 0302 clinical medicine, law, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Intensive Care Unit Admission, Simplexvirus, 030212 general & internal medicine, Simplified Acute Physiology Score, Acute Respiratory Distress Syndrome, MESH: Cohort Studies, MESH: Aged, 0303 health sciences, Univariate analysis, MESH: Middle Aged, medicine.diagnostic_test, Coinfection, Pneumonia, Pneumocystis, General Medicine, Middle Aged, Intensive care unit, 3. Good health, Infectious Diseases, Cytomegalovirus Infections, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Female, Bronchoalveolar Lavage Fluid, Human Immunodeficiency Virus, Microbiology (medical), medicine.medical_specialty, MESH: Cytomegalovirus, Intensive Care Unit, Article, 03 medical and health sciences, Internal medicine, medicine, Humans, MESH: Simplexvirus, Aged, Retrospective Studies, MESH: Humans, 030306 microbiology, business.industry, MESH: Bronchoalveolar Lavage Fluid, Retrospective cohort study, Herpes Simplex, MESH: Retrospective Studies, Odds ratio, MESH: Cytomegalovirus Infections, medicine.disease, MESH: Male, Herpes Simplex Virus, MESH: Coinfection, Pneumonia, Bronchoalveolar lavage, Immunology, business, MESH: Female, MESH: Pneumonia, Pneumocystis
Abstract: International audience; The purpose of this investigation was to characterize the management and prognosis of severe Pneumocystis jirovecii pneumonia (PJP) in human immunodeficiency virus (HIV)-negative patients. An observational cohort study of HIV-negative adults with PJP documented by bronchoalveolar lavage (BAL) through Gomori-Grocott staining or immunofluorescence, admitted to one intensive care unit (ICU) for acute respiratory failure, was undertaken. From 1990 to 2010, 70 patients (24 females, 46 males) were included, with a mean age of 58.6 ± 18.3 years. The mean Simplified Acute Physiology Score (SAPS)-II was 36.9 ± 20.4. Underlying conditions included hematologic malignancies (n = 21), vasculitis (n = 13), and solid tumors (n = 13). Most patients were receiving systemic corticosteroids (n = 63) and cytotoxic drugs (n = 51). Not a single patient received trimethoprim-sulfamethoxazole as PJP prophylaxis. Endotracheal intubation (ETI) was required in 42 patients (60.0 %), including 38 with acute respiratory distress syndrome (ARDS). In-ICU mortality was 52.9 % overall, reaching 80.9 % and 86.8 %, respectively, for patients who required ETI and for patients with ARDS. In the univariate analysis, in-ICU mortality was associated with SAPS-II (p = 0.0131), ARDS (p
Published: 2013
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15. Insight into the structure of the pUL89 C-terminal domain of the human cytomegalovirus terminase complex

Author: Couvreux, S, Hantz, S., Marquant, R., Champier, G., Alain, S., Morellet, N., Bouaziz, S., Couvreux, A., Teaching hospital, CHU Limoges, Ecosystèmes forestiers (UR EFNO), Institut national de recherche en sciences et technologies pour l'environnement et l'agriculture (IRSTEA), Institut d'électronique fondamentale (IEF), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Raherison, Sophie, Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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)-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)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-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), 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)-Institut de Chimie du CNRS (INC), Centre National de la Recherche Scientifique (CNRS) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Université Paris Descartes - Paris 5 (UPD5) - 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), and Université de Limoges (UNILIM) - Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503) - Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Models, Molecular, Magnetic Resonance Spectroscopy, MESH : Molecular Sequence Data, MESH : DNA, Cytomegalovirus, MESH: Protein Structure, Secondary, MESH: Amino Acid Sequence, Protein Structure, Secondary, MESH: Protein Structure, Tertiary, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Endonucleases, MESH : RNA, MESH : Endonucleases, MESH : Viral Proteins, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH : Sequence Alignment, MESH : Amino Acid Sequence, MESH: DNA, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH : Protein Structure, Tertiary, MESH: Biocatalysis, MESH: Models, Molecular, MESH: Cytomegalovirus, MESH : Models, Molecular, Molecular Sequence Data, MESH: Sequence Alignment, MESH : Endodeoxyribonucleases, MESH : Cytomegalovirus, Viral Proteins, MESH: RNA, Humans, Amino Acid Sequence, MESH: Endodeoxyribonucleases, Endodeoxyribonucleases, MESH: Humans, MESH: Molecular Sequence Data, MESH: Magnetic Resonance Spectroscopy, MESH : Humans, DNA, Endonucleases, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: Viral Proteins, Protein Structure, Tertiary, Biocatalysis, RNA, MESH : Magnetic Resonance Spectroscopy, MESH : Protein Structure, Secondary, MESH : Biocatalysis, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Sequence Alignment
Abstract: International audience; In a previous study, we identified 12 conserved domains within pUL89, the small terminase subunit of the human cytomegalovirus. A latter study showed that the fragment pUL89(580-600) plays an important role in the formation of the terminase complex by interacting with the large terminase subunit pUL56. In this study, analysis was performed to solve the structure of pUL89(568-635) in 50% H2O/50% acetonitrile (v/v). We showed that pUL89(568-635) consists of four alpha helices, but we did not identify any tertiary structure. The fragment 580-600 formed an amphipathic alpha helix, which had a hydrophobic side highly conserved among herpesviral homologs of pUL89; this was not observed for its hydrophilic side. The modeling of pUL89(457-612) using the recognition fold method allowed us to position pUL89(580-600) within this domain. The theoretical structure highlighted three important features. First, we identified a metal-binding pocket containing residues Asp(463), Glu(534), and Glu(588), which are highly conserved among pUL89 homologs. Second, the model predicted a positively charged surface able to interact with the DNA duplex during the nicking event. Third, a hydrophobic patch on the top of the catalytic site suggested that this may constitute part of the pUL89 site recognized by pUL56 potentially involved in DNA binding.
Published: 2010
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16. Structural bases for the affinity-driven selection of a public TCR against a dominant human cytomegalovirus epitope

Author: Xavier Saulquin, E. Debeaupuis, François Legoux, Madalen Le Gorrec, Adrien Kissenpfennig, Paul Machillot, B. Neveu, Jean-Baptiste Reiser, Marc Bonneville, Stephanie Gras, Dominique Housset, Nicole M. Thielens, Bernard Malissen, Anne Chouquet, Klara Echasserieau, Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-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)-Centre National de la Recherche Scientifique (CNRS), Recherches en cancérologie, Université de Nantes (UN)-IFR26-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre d'Immunologie de Marseille - Luminy (CIML), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)
Subjects: Human cytomegalovirus, Amino Acid Motifs, Cytomegalovirus, Epitopes, T-Lymphocyte, Crystallography, X-Ray, Lymphocyte Activation, Epitope, MESH: Virus Activation, MESH: Amino Acid Motifs, 0302 clinical medicine, Immunology and Allergy, Cytotoxic T cell, Medicine(all), Genetics, 0303 health sciences, MESH: Peptides, MESH: Receptors, Antigen, T-Cell, medicine.anatomical_structure, Cytomegalovirus Infections, [SDV.IMM]Life Sciences [q-bio]/Immunology, Protein Binding, MESH: Cytomegalovirus, T cell, Immunology, Receptors, Antigen, T-Cell, MESH: Immunodominant Epitopes, chemical and pharmacologic phenomena, Biology, Major histocompatibility complex, MESH: Phosphoproteins, Viral Matrix Proteins, MESH: Epitopes, T-Lymphocyte, 03 medical and health sciences, Antigen, HLA-A2 Antigen, medicine, Humans, MESH: Protein Binding, MESH: Lymphocyte Activation, MESH: HLA-A Antigens, 030304 developmental biology, MESH: Viral Matrix Proteins, MESH: Humans, HLA-A Antigens, Immunodominant Epitopes, T-cell receptor, MESH: Clone Cells, MESH: Cytomegalovirus Infections, Phosphoproteins, medicine.disease, MESH: Crystallography, X-Ray, Clone Cells, Transplantation, biology.protein, Virus Activation, Peptides, 030215 immunology
Abstract: Protective T cell responses elicited along chronic human CMV (HCMV) infections are sometimes dominated by CD8 T cell clones bearing highly related or identical public TCR in unrelated individuals. To understand the principles that guide emergence of these public T cell responses, we have performed structural, biophysical, and functional analyses of an immunodominant public TCR (RA14) directed against a major HLA-A*0201-restricted HCMV Ag (pp65495–503) and selected in vivo from a diverse repertoire after chronic stimulations. Unlike the two immunodominant public TCRs crystallized so far, which focused on one peptide hotspot, the HCMV-specific RA14 TCR interacts with the full array of available peptide residues. The conservation of some peptide-MHC complex-contacting amino acids by lower-affinity TCRs suggests a shared TCR-peptide-MHC complex docking mode and supports an Ag-driven selection of optimal TCRs. Therefore, the emergence of a public TCR of an oligoclonal Ag-specific response after repeated viral stimulations is based on a receptor displaying a high structural complementarity with the entire peptide and focusing on three peptide hotspots. This highlights key parameters underlying the selection of a protective T cell response against HCMV infection, which remains a major health issue in patients undergoing bone marrow transplantation.
Published: 2009
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17. Bcl-3-regulated transcription from major immediate-early promoter of human cytomegalovirus in monocyte-derived macrophages

Author: Kashif Aziz Khan, Alain Coaquette, Christian Davrinche, Georges Herbein, Department of Pathology, University of Veterinary and Animal Sciences, Lahore, Departement /u563 : Immunologie et Maladies infectieuses, 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)-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), Agents pathogènes et inflammation - UFC (EA 4266) (API), Université de Franche-Comté (UFC), and Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)
Subjects: Human cytomegalovirus, Time Factors, Transcription, Genetic, viruses, MESH: NF-kappa B, Cytomegalovirus, Virus Replication, Transactivation, 0302 clinical medicine, B-Cell Lymphoma 3 Protein, MESH: RNA, Small Interfering, MESH: Up-Regulation, Immunology and Allergy, RNA, Small Interfering, Cells, Cultured, 0303 health sciences, U937 cell, NF-kappa B, Transfection, MESH: Transcription Factors, MESH: Gene Expression Regulation, 3. Good health, Up-Regulation, 030220 oncology & carcinogenesis, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Cells, Cultured, Protein Binding, MESH: Cytomegalovirus, Immunology, Biology, 03 medical and health sciences, Proto-Oncogene Proteins, medicine, MESH: Protein Binding, Humans, Luciferase, 030304 developmental biology, MESH: Humans, MESH: Transcription, Genetic, Macrophages, MESH: Time Factors, MESH: Virus Replication, MESH: Macrophages, Fibroblasts, medicine.disease, Molecular biology, In vitro, MESH: Proto-Oncogene Proteins, Viral replication, Gene Expression Regulation, MESH: Fibroblasts, Chromatin immunoprecipitation, Transcription Factors
Abstract: Monocytes/macrophages are key cells in the pathogenesis of human CMV (HCMV) infection, but the in vitro rate of viral production in primary human monocyte-derived macrophages (MDM) is considerably lower than in fibroblasts. Considering that the NF-κB signaling pathway is potentially involved in the replication strategy of HCMV through efficient transactivation of the major immediate-early promoter (MIEP), efficient viral replication, and late gene expression, we investigated the composition of the NF-κB complex in HCMV-infected MDMs and fibroblasts. Preliminary studies showed that HCMV could grow in primary MDM culture but that the viral titer in culture supernatants was lower than that observed in the supernatants of more permissive MRC5 fibroblasts. EMSA and microwell colorimetric NF-κB assay demonstrated that HCMV infection of MDMs increased p52 binding activity without activating the canonical p50/p65 complex. Moreover, Bcl-3 was up-regulated and was demonstrated to associate with p52, indicating p52/Bcl-3 complexes as the major component of the NF-κB complex in MDMs. Luciferase assays in promonocytic U937 cells transfected with an MIEP-luciferase reporter construct demonstrated MIEP activation in response to p52 and Bcl-3 overexpression. Chromatin immunoprecipitation assay demonstrated that p52 and Bcl-3 bind the MIEP in acutely HCMV-infected MDMs. In contrast, HCMV infection of MRC5 fibroblasts resulted in activation of p50/p65 heterodimers. Thus, activation of p52/Bcl-3 complexes in MDMs and p50/p65 heterodimers in fibroblasts in response to HCMV infection might explain the low-level growth of the virus in MDMs vs efficient growth in fibroblasts.
Published: 2009
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18. Cytomegalovirus microRNAs

Author: Lars Dölken, Sébastien Pfeffer, Ulrich H. Koszinowski, Max Von Pettenkofer Institute (MVP), Ludwig-Maximilians-Universität München (LMU), Architecture et Réactivité de l'ARN (ARN), Institut de biologie moléculaire et cellulaire (IBMC), and Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Gene Expression Regulation, Viral, MESH: Cytomegalovirus, Pan troglodytes, viruses, Cytomegalovirus, MESH: Pan troglodytes, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Mice, 03 medical and health sciences, MESH: Gene Expression Regulation, Viral, 0302 clinical medicine, Virology, Genetics, Animals, Humans, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecular Biology, MESH: Mice, HCMV, 030304 developmental biology, miRNA, 0303 health sciences, MESH: Humans, CCMV, virus diseases, Herpesvirus, General Medicine, MESH: Cytomegalovirus Infections, 3. Good health, MicroRNAs, 030220 oncology & carcinogenesis, Cytomegalovirus Infections, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: MicroRNAs, MCMV
Abstract: International audience; MicroRNAs (miRNAs) are small, non-coding RNA molecules that regulate gene expression at a post-transcriptional level in virtually all eukaryotic organisms and some viruses, particularly herpesviruses. miRNAs are non-immunogenic, stealthy tools for viruses to regulate their as well as host gene expression. The human cytomegalovirus (HCMV) is the major cause of morbidity in immunocompromised patients and allogenic bone-marrow or organ-transplant recipients and the leading cause of congenital birth defects. HCMV miRNAs may provide valuable targets for new urgently needed antiviral drugs. This review focuses on recent findings for viral miRNAs expressed by cytomegaloviruses (CMV) including data from human, chimpanzee, and murine CMV. These are discussed in the context of findings for other viruses to highlight potentially conserved roles exerted by viral miRNAs.
Published: 2009
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19. Direct genotyping of cytomegalovirus envelope glycoproteins from toddler's saliva samples

Author: Jérôme Grosjean, P. Brosset, François Denis, Sébastien Hantz, Sophie Alain, F. Undreiner, C. Pasquier, L. Trapes, B. Virey, Marie-Claire Baclet, Catherine Mengelle, Sébastien Cotin, Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, 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), Service de Pédiatrie médicale [CHU Limoges], Association Française de Pédiatrie Ambulatoire (AFPA), and Raherison, Sophie
Subjects: Human cytomegalovirus, Saliva, MESH: Child Day Care Centers, Cytomegalovirus, Polymerase Chain Reaction, law.invention, MESH: Genotype, 0302 clinical medicine, Viral Envelope Proteins, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, law, Genotype, 030212 general & internal medicine, Polymerase chain reaction, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Infectivity, Genetics, 0303 health sciences, MESH: Polymorphism, Single Nucleotide, MESH: Infant, 3. Good health, Infectious Diseases, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Cytomegalovirus Infections, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Restriction fragment length polymorphism, Polymorphism, Restriction Fragment Length, MESH: Cytomegalovirus, Biology, Polymorphism, Single Nucleotide, 03 medical and health sciences, Viral Proteins, Virology, medicine, Humans, MESH: Saliva, Typing, Genotyping, 030304 developmental biology, MESH: Humans, MESH: Polymorphism, Restriction Fragment Length, Infant, MESH: Polymerase Chain Reaction, MESH: Cytomegalovirus Infections, Child Day Care Centers, medicine.disease, MESH: Viral Proteins, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: Viral Envelope Proteins, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
Abstract: Centre Hospitalier Universitaire (CHU) Limoges, Centre National de Reference du CMV, Laboratoire de Bactériologie-Virologie-Hygiène, Limoges, France; International audience; BACKGROUND: The polymorphism of genes encoding CMV envelope protein is used for strain classification and may influence pathogenesis and/or infectivity. CMV genotyping is usually based on sequencing or acrylamide gel-RFLP, but these methods are not suited to rapid screening of large populations. OBJECTIVES: We developed a high-throughput method to analyze CMV strains diversity and to detect multiple-strain infection in a large population of toddlers (six daycare centers (DCC) and an emergency unit (EU)). METHODS: We developed a new PCR-RFLP method coupled with capillary electrophoresis fragment detection for UL55-gB, UL75-gH and UL73-gN genotyping. To detect gB recombinants, gpUL55 typing was applied to two variable zones (NTerminal and central). We applied this method to 212 CMV-positive saliva samples and controlled the results by direct sequencing of PCR products. RESULTS: We identified 112 strains, that fell into eight groups in UL55-gB, two groups in UL75-gH, and seven groups in UL73-gN. The 79 samples from the emergency unit contained 30 strains, 28 children harboring 2 strains. The samples (n=133) from the six daycare centers contained respectively 4, 1, 6, 1 and 11 strains. Fifteen percent of strains were UL55-gB recombinants. CONCLUSION: Our new method can simultaneously determine gB, gH and gN genotypes and offers more precise classification of CMV strains than previous RFLP-based methods. This could constitute the basis for a new classification, particularly in UL55-gB. Easy direct identification of multiple strains and recombinants in pathological samples could facilitate large epidemiologic studies.
Published: 2009
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20. Rapid determination of antiviral drug susceptibility of human cytomegalovirus by real-time PCR

Author: Marie-Christine Mazeron, Marie-José Sanson-Le Pors, Françoise Petit, Sophie Alain, Nicolas Boiteau, Nathalie Schnepf, Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Limoges (UNILIM), Hôpital Lariboisière, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Lariboisière-Université Paris Diderot - Paris 7 (UPD7), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Lariboisière-Fernand-Widal [APHP], and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)
Subjects: Foscarnet, Human cytomegalovirus, viruses, Cytomegalovirus, MESH: DNA Replication, Viral Plaque Assay, Polymerase Chain Reaction, chemistry.chemical_compound, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Cells, Cultured, 0303 health sciences, virus diseases, 3. Good health, MESH: Reproducibility of Results, Real-time polymerase chain reaction, Cytomegalovirus Infections, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, medicine.drug, Cidofovir, MESH: Cells, Cultured, DNA Replication, Ganciclovir, MESH: Antiviral Agents, MESH: Plaque Assay, MESH: Cytomegalovirus, medicine.drug_class, Congenital cytomegalovirus infection, Biology, Antiviral Agents, 03 medical and health sciences, Virology, medicine, Humans, 030304 developmental biology, Pharmacology, MESH: Humans, 030306 microbiology, Reproducibility of Results, MESH: Polymerase Chain Reaction, MESH: Cytomegalovirus Infections, biochemical phenomena, metabolism, and nutrition, medicine.disease, Molecular biology, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: DNA, Viral, chemistry, DNA, Viral, Antiviral drug
Abstract: International audience; A quantitative real-time PCR-based assay was developed for determination of cytomegalovirus (HCMV) susceptibility to antiviral drugs. After HCMV isolate-growth for 4 days, antiviral drug susceptibility was determined by measuring the reduction of intracellular HCMV DNA in the presence of increasing concentrations of either ganciclovir, or foscarnet or cidofovir. The 50% inhibitory concentration (IC(50)) was the drug concentration that reduced the number of HCMV genome copies by 50%. The IC(50) values were measured for seven HCMV reference strains sensitive or resistant to one or more antiviral drugs. The antiviral susceptibility of 21 HCMV isolates was then tested and the results were consistent with prior determination of their phenotype and/or genotype by plaque reduction assay and sequencing. The real-time PCR susceptibility assay reported here was found to be highly reproducible, simpler to perform than the plaque reduction assay, and amenable to use in the routine diagnostic virology laboratory.
Published: 2009
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21. Early immune response against retrovirally transduced herpes simplex virus thymidine kinase-expressing gene-modified T cells coinfused with a T cell-depleted marrow graft: an altered immune response?

Author: Christophe Ferrand, Jean Marie Certoux, Bruno Lioure, Delphine Sauce, Patricia Mercier-Letondal, Pierre Tiberghien, Eric Deconinck, Eric Robinet, Noel Milpied, Marina Deschamps, Jean-Yves Cahn, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Plateforme de Biomonitoring, Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté]), Service des maladies du sang, CHU Bordeaux [Bordeaux]-Hôpital Haut-Lévêque [CHU Bordeaux], CHU Bordeaux [Bordeaux]-Groupe Hospitalier Sud, Service d'onco-hématologie, CHU Strasbourg-Hôpital de Hautepierre [Strasbourg], Hematology, Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble, Service greffe de moelle osseuse, Université Paris Diderot - Paris 7 (UPD7)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Laboratoire de biologie moléculaire, EFS, Saas, Philippe, Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], CHU Bordeaux [Bordeaux]-Groupe Hospitalier Sud-Hôpital Haut-Lévêque [CHU Bordeaux], and CHU Bordeaux [Bordeaux]
Subjects: Cytotoxicity, Immunologic, Herpesvirus 4, Human, Genetic enhancement, T-Lymphocytes, Cytomegalovirus, Graft vs Host Disease, MESH: Neomycin, MESH: Amino Acid Sequence, 0302 clinical medicine, Interferon, Transduction, Genetic, MESH: Genetic Vectors, Cytotoxic T cell, Simplexvirus, MESH: Bone Marrow Transplantation, Bone Marrow Transplantation, 0303 health sciences, MESH: Middle Aged, Middle Aged, MESH: Transduction, Genetic, 3. Good health, medicine.anatomical_structure, surgical procedures, operative, MESH: Retroviridae, 030220 oncology & carcinogenesis, Molecular Medicine, [SDV.IMM]Life Sciences [q-bio]/Immunology, medicine.drug, Adult, MESH: Thymidine Kinase, MESH: Cytomegalovirus, MESH: Lymphocyte Count, [SDV.IMM] Life Sciences [q-bio]/Immunology, CD3, T cell, Genetic Vectors, MESH: Graft vs Host Disease, chemical and pharmacologic phenomena, Biology, In Vitro Techniques, Thymidine Kinase, Lymphocyte Depletion, 03 medical and health sciences, Viral Proteins, Immune system, Genetics, medicine, MESH: Transplantation, Homologous, Humans, Transplantation, Homologous, MESH: Simplexvirus, Amino Acid Sequence, Lymphocyte Count, MESH: Cytotoxicity, Immunologic, Molecular Biology, 030304 developmental biology, MESH: Lymphocyte Depletion, MESH: Humans, Neomycin, MESH: Adult, MESH: Herpesvirus 4, Human, MESH: Viral Proteins, Retroviridae, MESH: T-Lymphocytes, Thymidine kinase, Immunology, biology.protein, Bone marrow
Abstract: International audience; Administration of herpes simplex thymidine kinase (HSV-tk)-expressing, gene-modified T cells (GMCs) with T cell-depleted bone marrow transplantation (TCD-BMT) can allow modulation of posttransplantation alloreactivity. Twelve patients received 2 x 10(5) to 2 x 10(6) CD3+ donor GMCs per kilogram with HLA-identical sibling TCD-BMT. Despite extensive T cell depletion of bone marrow, an intensive conditioning regimen, and immunosuppressive graft-versus-host disease (GvHD) prophylaxis, infusion at the time of TCD-BMT of this low number of GMCs sufficed to induce a rapid GMC-specific immune response, as detected by interferon- enzyme- linked immunospot assay in six of eight patients, preferentially targeting HSV-tk. Maximal responses were reached early (median time, 49 [35-68] days post-BMT), with a subsequent rapid and significant decrease in five of six evaluable patients. Immune responses were negatively correlated with the maximal circulating GMC counts. However, such immune response did not result in the elimination of circulating GMCs and was not associated with measurable ex vivo cytotoxic activity against GMCs. Furthermore, alloreactive GMCs still could induce GCV-sensitive GvHD in one patient despite an ongoing immune response. Overall, infusion of HSV-tk-expressing GMCs at the time of BMT results in an early immune response. Such immune response may be altered and may not prevent persistent GCV-sensitive alloreactivity.
Published: 2008

22. Replication and packaging of an infectious bursal disease virus segment A-derived minigenome

Author: Bernard Delmas, Helmi Mardassi, N. Ben Abdeljelil, Institut Pasteur de Tunis, Réseau International des Instituts Pasteur (RIIP), Unité de recherche Virologie et Immunologie Moléculaires (VIM (UR 0892)), Institut National de la Recherche Agronomique (INRA), and Unité de recherche Virologie et Immunologie Moléculaires (VIM)
Subjects: Cancer Research, Cytomegalovirus, Virus Replication, Infectious bursal disease virus, Infectious bursal disease, Green fluorescent protein, chemistry.chemical_compound, Plasmid, Genes, Reporter, RNA polymerase, Chlorocebus aethiops, MESH: Animals, Viral, Promoter Regions, Genetic, Catalytic, 0303 health sciences, Genome, biology, MESH: Infectious bursal disease virus, Ribozyme, MESH: COS Cells, RNA silencing, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Infectious Diseases, MESH: RNA, Viral, COS Cells, RNA, Viral, MESH: Genome, Viral, Hepatitis Delta Virus, MESH: Cytomegalovirus, animal structures, MESH: Virus Assembly, Green Fluorescent Proteins, MESH: Vero Cells, Genome, Viral, Virus, Cercopithecus aethiops, Promoter Regions, 03 medical and health sciences, MESH: Green Fluorescent Proteins, Genetic, Virology, MESH: RNA, Catalytic, MESH: Promoter Regions, Genetic, medicine, Animals, RNA, Catalytic, Gene, Reporter, Vero Cells, 030304 developmental biology, 030306 microbiology, Virus Assembly, MESH: Virus Replication, MESH: Genes, Reporter, medicine.disease, Molecular biology, MESH: Cercopithecus aethiops, MESH: Hepatitis Delta Virus, chemistry, Genes, biology.protein, RNA
Abstract: International audience; A synthetic cytomegalovirus (CMV) promoter-driven cDNA minigenome containing the enhanced green fluorescence protein (EGFP) gene as a reporter was derived from the genomic segment A of infectious bursal disease virus (IBDV). The 5'-end of the minigenome was fused to the transcription start site of the immediate early CMV promoter, and the hepatitis delta virus (HDV) ribozyme sequence was added at its 3'-end. We show that co-transfection of the minigenome with a plasmid encoding the IBDV RNA-dependent RNA polymerase VP1, results in a consistent increase of the EGFP expression, as measured by fluorescence microscopy and flow cytometry assays. Replication of the minigenome-derived transcript was evidenced by real-time RT-PCR analyses targeted to both the plus- and minus-sense strands. When cells were infected with IBDV and transfected with the plasmid carrying the minigenome, the minigenome was packaged and EGFP was found to be expressed in a second cycle of infection. These results show the potential use of this system as a new tool to characterize IBDV replication and genome packaging.
Published: 2008
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23. Putative functional domains of human cytomegalovirus pUL56 involved in dimerization and benzimidazole D-ribonucleoside activity

Author: Armelle Rametti, Serge Bouaziz, François Denis, Sébastien Hantz, Sophie Alain, Marie-Christine Mazeron, Anthony Couvreux, Gaël Champier, Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Limoges (UNILIM), Hôpital Lariboisière, Université Paris Diderot - Paris 7 (UPD7)-Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-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), 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)-Institut de Chimie du CNRS (INC), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, Laboratoire de Biochimie Pharmaceutique, Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Laboratoire d'informatique Fondamentale de Marseille - UMR 6166 (LIF), Université de la Méditerranée - Aix-Marseille 2-Université de Provence - Aix-Marseille 1-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Lariboisière-Université Paris Diderot - Paris 7 (UPD7), Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-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)-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)-Centre National de la Recherche Scientifique (CNRS), Bouaziz, Serge, Université de Rennes (UR), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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)-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)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Limoges (UNILIM) - Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503) - Institut National de la Santé et de la Recherche Médicale (INSERM), Assistance publique - Hôpitaux de Paris (AP-HP) - Hôpital Lariboisière - Université Paris Diderot - Paris 7 (UPD7), Centre National de la Recherche Scientifique (CNRS) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Université Paris Descartes - Paris 5 (UPD5) - Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), and 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)
Subjects: Human cytomegalovirus, Models, Molecular, MESH: Sequence Analysis, DNA, MESH : Molecular Sequence Data, viruses, Cytomegalovirus, MESH: Amino Acid Sequence, medicine.disease_cause, MESH : Ribonucleosides, chemistry.chemical_compound, Letermovir, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Pharmacology (medical), MESH : DNA, Viral, Cells, Cultured, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], MESH : Amino Acid Sequence, MESH : Viral Structural Proteins, virus diseases, Infectious Diseases, MESH : Benzimidazoles, Biochemistry, MESH : Dimerization, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Dna packaging, Dimerization, MESH: Models, Molecular, MESH : Virus Assembly, medicine.drug, MESH: Cells, Cultured, Benzimidazole, MESH: Cytomegalovirus, MESH: Virus Assembly, MESH : Models, Molecular, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Molecular Sequence Data, MESH: Viral Structural Proteins, Virus, Herpesviridae, MESH : Cytomegalovirus, Betaherpesvirinae, MESH : Cells, Cultured, MESH : Fibroblasts, medicine, Humans, Amino Acid Sequence, Pharmacology, Viral Structural Proteins, MESH: Ribonucleosides, MESH: Humans, MESH: Molecular Sequence Data, Virus Assembly, MESH : Humans, Sequence Analysis, DNA, Fibroblasts, biochemical phenomena, metabolism, and nutrition, Ribonucleoside, biology.organism_classification, medicine.disease, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: DNA, Viral, chemistry, MESH: Dimerization, MESH: Fibroblasts, DNA, Viral, Benzimidazoles, Ribonucleosides, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: Benzimidazoles, MESH : Sequence Analysis, DNA
Abstract: Background Benzimidazole d-ribonucleosides inhibit DNA packaging during human cytomegalovirus (HCMV) replication. Although they have been shown to target pUL56 and pUL89 (the large and small subunits of the HCMV terminase, respectively) their mechanism of action is not yet fully understood. We aimed here to better understand HCMV DNA maturation and the mechanism of action of benzimidazole derivatives. Methods The HCMV pUL56 protein was studied by sequence analysis of the HCMV UL56 gene and herpesvirus counterparts combined with primary structure analysis of the corresponding amino acid sequences. Results The UL56 sequence analysis of 45 HCMV strains and counterparts among herpesviruses allowed the identification of 12 conserved regions. Moreover, comparison with the product of gene 49 (gp49) of bacteriophage T4 suggested that the pUL56 zinc finger is localized close to the dimerization site of pUL56, providing a spatial organization of the catalytic site that allows recognition and cleavage of DNA. Conclusions This study provides a basis to investigate the mechanism of concatemeric DNA cleavage and a biochemical basis for DNA packaging inhibition by benzimidazole derivatives.
Published: 2008

24. Maintenance of HIV-specific central and effector memory CD4 and CD8 T cells requires antigen persistence

Author: Gennady Bocharov, Urban Sester, Hans Köhler, Barbara Gärtner, Martina Sester, Hans W. Pees, Andreas Meyerhans, Simon Wain-Hobson, Biologie et Gestion des Adventices ( BGA ), Etablissement National d'Enseignement Supérieur Agronomique de Dijon ( ENESAD ) -Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ), Rétrovirologie Moléculaire, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Biologie et Gestion des Adventices (BGA), Université de Bourgogne (UB)-Institut National de la Recherche Agronomique (INRA)-Etablissement National d'Enseignement Supérieur Agronomique de Dijon (ENESAD), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Etablissement National d'Enseignement Supérieur Agronomique de Dijon (ENESAD)-Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)
Subjects: CD4-Positive T-Lymphocytes, Cytotoxic, T-Lymphocytes, Cytomegalovirus, HIV Infections, MESH : Viral Load, MESH: T-Lymphocyte Subsets, CD8-Positive T-Lymphocytes, MESH: Antiretroviral Therapy, Highly Active, MESH: HIV-1, 0302 clinical medicine, T-Lymphocyte Subsets, Antiretroviral Therapy, Highly Active, Cytotoxic T cell, 0303 health sciences, biology, MESH: Kinetics, Effector, virus diseases, MESH: CD4-Positive T-Lymphocytes, MESH: HIV Infections, Viral Load, MESH : CD8-Positive T-Lymphocytes, MESH: CD8-Positive T-Lymphocytes, 3. Good health, Infectious Diseases, medicine.anatomical_structure, MESH : CD4-Positive T-Lymphocytes, Lentivirus, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Immunologic Memory, MESH : Kinetics, MESH: Viral Load, MESH : HIV-1, MESH: Cytomegalovirus, MESH : Immunologic Memory, T cell, Immunology, Antiretroviral Therapy, [ SDV.MP.VIR ] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH : T-Lymphocytes, Cytotoxic, MESH : Cytomegalovirus, 03 medical and health sciences, Antigen, Virology, medicine, MESH : HIV Infections, Humans, Highly Active, 030304 developmental biology, MESH: Humans, MESH : Humans, T lymphocyte, biology.organism_classification, MESH : Antiretroviral Therapy, Highly Active, MESH : T-Lymphocyte Subsets, Kinetics, HIV-1, Memory T cell, Immunologic Memory, MESH: T-Lymphocytes, Cytotoxic, CD8, T-Lymphocytes, Cytotoxic, 030215 immunology
Abstract: International audience; The HIV-specific central and effector CD4 and CD8 memory T cell populations disappear from the peripheral blood of infected individuals under highly active antiretroviral therapy (HAART) with a mean half-life of 6.0 and 7.7 months, respectively. By contrast, cytomegalovirus (CMV)-specific responses are stable or increase. The striking quantitative differences between T cell memory to two persistent viral infections are instructive as to how antigen dosage contributes to the maintenance of antigen-specific memory T cell responses in humans.
Published: 2007
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25. [Cytomegalovirus and cryptogenic inflammatory bowel disease]

Author: Pofelski, Joanna, Heluwaert, Frédéric, Roblin, Xavier, Morand, Patrice, Gratacap, Bénédicte, Germain, Emmanuel, Brion, Jean-Paul, Salon, Caroline, Bonaz, Bruno, Sinniger, Valérie, Service d'hépato-gastroentérologie, CHU Grenoble, Laboratoire de virologie moléculaire et structurale, Département de médecine aiguë spécialisée, CHU Grenoble-Hôpital Michallon, and Département d'anatomo-pathologie
Subjects: MESH: Antiviral Agents, MESH: Cytomegalovirus, MESH: Humans, MESH: Immunologic Factors, MESH: Virus Replication, Cytomegalovirus, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, MESH: Cytomegalovirus Infections, MESH: Inflammatory Bowel Diseases, Inflammatory Bowel Diseases, Virus Replication, digestive system, Antiviral Agents, digestive system diseases, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Cytomegalovirus Infections, Humans, Immunologic Factors
Abstract: International audience; CMV infection has been reported in association with some flares of IBD. Its prevalence varies with the method of diagnosis and the severity of IBD. Although the link between CMV and IBD is not clear, the immunomodulator properties of the virus may play a role in the evolution of IBD. Besides the necessity of immunosuppression to treat IBD, inflammation per se can maintain in situ viral replication. Antiviral treatment can be useful in some situations. New molecular methods will permit earlier and more sensitive diagnosis of CMV infection and a better evaluation of treatment efficacy.
Published: 2007

26. Monitoring human cytomegalovirus (HCMV) in HCMV-seropositive orthotopic liver-transplant recipients by means of quantitative real-time polymerase chain reaction

Author: Mengelle, Catherine, Legrand-Abravanel, Florence, Kamar, Nassim, Alain, Sophie, Basse, Grégoire, Pillet, Adèle, Lavayssière, Laurence, Suc, Bertrand, Izopet, Jacques, Rostaing, Lionel, Université de Limoges (UNILIM), Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Bucek, Ingrid
Subjects: Graft Rejection, MESH: Cytomegalovirus, MESH: Liver Transplantation, Time Factors, Cytomegalovirus, MESH: Graft Rejection, MESH: Creatinine, Polymerase Chain Reaction, Postoperative Complications, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Postoperative Complications, Humans, Retrospective Studies, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Humans, MESH: Time Factors, virus diseases, MESH: Polymerase Chain Reaction, MESH: Retrospective Studies, MESH: Cytomegalovirus Infections, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Liver Transplantation, Creatinine, Cytomegalovirus Infections, Epidemiological Monitoring, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: Environmental Monitoring, Environmental Monitoring
Abstract: International audience; OBJECTIVE: Human cytomegalovirus can be reactivated after orthotopic liver transplantation in patients who are seropositive for cytomegalovirus. Whether those cytomegalovirus-seropositive patients require immediate posttransplant (anti)- cytomegalovirus prophylactic therapy or preemptive treatment as opposed to deferred treatment remains controversial. The aims of our study were to evaluate the relevance of cytomegalovirus monitoring with quantitative real-time polymerase chain reaction in whole blood and to analyze the factors that determine the treatment of the first episode of cytomegalovirus infection with intravenous ganciclovir in seropositive liver-transplant patients. PATIENTS AND METHODS: Forty-two cytomegalovirus-seropositive liver-transplant patients were assessed for cytomegalovirus DNAemia every 2 weeks until posttransplant day 90 and every 3 to 4 weeks until day 180. Biochemical and hematologic parameters were also prospectively monitored. RESULTS: Cytomegalovirus DNAemia was detected at least once in 27 patients (64%). Treatment was initiated in 12 patients (group 1) but not in 15 others (group 2). Median HCMV viral loads of the first positive and the highest DNAemia were statistically higher in group 1 than in group 2 (P = 0.01). Univariate analysis of DNAemia showed that alkaline phosphatase levels were significantly higher in group 1 than in group 2 (P = .0011) and that hemoglobin levels were significantly lower in group 1 than in group 2 (P = .0443). The results of multivariate analysis showed that the only factor that predicted the treatment of the first episode of HCMV DNAemia was a level of alkaline phosphatase greater than 150 IU/L [odds ratio, 20; range, 1.97-203.32; P = .01]. CONCLUSIONS: A combination of criteria, including viral-load kinetics, clinical factors, alkaline phosphatase levels (in particular), and the patient's immune condition, is required to efficiently monitor patients who are seropositive for cytomegalovirus after orthotopic liver transplantation.
Published: 2006

27. Multiplex real-time PCR assay for simultaneous quantitation of human cytomegalovirus and herpesvirus-6 in polymorphonuclear and mononuclear cells of transplant recipients

Author: Jean-Christophe Szelag, François Denis, Ludovic Couty, Yann Le Meur, Florence Rolle, Karine Pradeau, Pascal Turlure, Pierre Ferrat, Sylvie Ranger-Rogez, Dominique Bordessoule, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service d'Hématologie clinique et thérapie cellulaire [CHU Limoges], CHU Limoges, Service d'Hématologie biologique [CHU Limoges], Université de Limoges (UNILIM), Physiologie Moléculaire de la Réponse Immune et des Lymphoproliférations (PMRIL), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Centre National de la Recherche Scientifique (CNRS), Biologie moléculaire et cellulaire des microorganismes (EA3175), and Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: Male, Human cytomegalovirus, MESH: Fluorescence Resonance Energy Transfer, Neutrophils, MESH: Oligonucleotide Probes, Herpesvirus 6, Human, viruses, Cytomegalovirus, MESH: Neutrophils, medicine.disease_cause, Polymerase Chain Reaction, Fluorescence Resonance Energy Transfer, Multiplex, Antigens, Viral, 0303 health sciences, MESH: Middle Aged, biology, virus diseases, MESH: Transplantation, Middle Aged, 3. Good health, MESH: Leukocytes, Mononuclear, Real-time polymerase chain reaction, [SDV.IMM]Life Sciences [q-bio]/Immunology, Female, Human herpesvirus 6, MESH: Antigens, Viral, Adult, MESH: Cytomegalovirus, Sensitivity and Specificity, Peripheral blood mononuclear cell, Herpesviridae, 03 medical and health sciences, Virology, Multiplex polymerase chain reaction, medicine, TaqMan, Humans, 030304 developmental biology, Transplantation, MESH: Humans, 030306 microbiology, MESH: Adult, MESH: Polymerase Chain Reaction, biochemical phenomena, metabolism, and nutrition, medicine.disease, biology.organism_classification, MESH: Herpesvirus 6, Human, Molecular biology, MESH: Sensitivity and Specificity, MESH: Male, MESH: DNA, Viral, DNA, Viral, Leukocytes, Mononuclear, Oligonucleotide Probes, MESH: Female
Abstract: International audience; Human cytomegalovirus (HCMV) and human herpesvirus-6 (HHV-6) are two closely related viruses, which belong to the Herpesviridae family. Following primary infection, they are thought to persist for life as latent forms in mononuclear cells. HCMV and HHV-6 can cause considerable morbidity in immunocompromised individuals, such as transplant patients. A sensitive and specific LightCycler multiplex real-time PCR assay based on fluorescence energy transfer (known as FRET) was developed. This assay, by using two sets of hybridization probes specific for HHV-6 (A and B) and HCMV, can differentiate reliably and quantify simultaneously both viruses in order to diagnose reactivation processes. The assay was optimized and the lower limit of detection for both viruses was determined to be 10 viral genome copies per reaction. Both viruses were quantified in 83 peripheral blood mononuclear cells (PBMCs) and 87 polymorphonuclear leukocytes (PMNLs) collected from 32 transplant recipients. This multiplex real-time quantitative PCR was finally compared with two other quantitation and detection assays used daily in laboratory (PCR DIG detection and antigenemia for HCMV, TaqMan Assay for HHV-6). This technique can be useful for the differentiation and quantitation of HCMV and HHV-6 for monitoring transplant patients.
Published: 2006
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28. Phenotypic characterisation of cytomegalovirus DNA polymerase: a method to study cytomegalovirus isolates resistant to foscarnet

Author: Marie-José Sanson-Le Pors, Jérôme Gravisse, Marie-Christine Mazeron, Anne-Marie Fillet, Françoise Petit, Sophie Alain, Alexandra Ducancelle, Hôpital Lariboisière, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Limoges (UNILIM), Service de virologie [CHU Pitié-Salpêtrière], Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Lariboisière-Université Paris Diderot - Paris 7 (UPD7), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Pitié-Salpêtrière [APHP]
Subjects: Human cytomegalovirus, Foscarnet, MESH: Cytomegalovirus, DNA polymerase, Cytomegalovirus, DNA-Directed DNA Polymerase, 03 medical and health sciences, chemistry.chemical_compound, Transcription (biology), [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Virology, Drug Resistance, Viral, MESH: DNA-Directed DNA Polymerase, medicine, MESH: Genetic Variation, Gene, Polymerase, 030304 developmental biology, 0303 health sciences, Expression vector, MESH: Drug Resistance, Viral, biology, 030306 microbiology, Genetic Variation, virus diseases, MESH: Cytomegalovirus Infections, biochemical phenomena, metabolism, and nutrition, medicine.disease, Molecular biology, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 3. Good health, chemistry, Cytomegalovirus Infections, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, biology.protein, DNA, medicine.drug, MESH: Foscarnet
Abstract: International audience; A phenotypic method was developed to test mutations in the human cytomegalovirus (HCMV) DNA polymerase gene (UL54) suspected to confer resistance to foscarnet. This method was used to determine the biochemical phenotype of wild-type and mutated HCMV DNA polymerases that had been synthesised in vitro as follows. The UL54 genes were amplified from foscarnet-resistant and -sensitive isolates by PCR and the products were cloned into an expression vector under the control of a T7 promoter. Mutations were introduced by site-directed mutagenesis into wild-type gene UL54 and then polymerases were synthesised by using a commercially available coupled transcription/translation system. Polymerase activity was measured with and without foscarnet by detecting the incorporation of digoxigenin-labelled nucleotides into the growing DNA chain. The results of this non-radioactive assay were consistent with those obtained with the conventional radioactive assay. It was found that the activity of polymerases containing the V715M or E756K mutations was inhibited by foscarnet at concentrations 70- and 30-fold higher than that of wild-type polymerase, respectively. Change N495K and a combination of changes K415R and S291P, both observed in foscarnet-resistant isolates, induced a 5- and 10-fold decrease in susceptibility to foscarnet, respectively. This non-radioactive phenotypic assay could be useful for the characterisation of mutations that confer HCMV resistance to foscarnet.
Published: 2005
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29. Assessment of estrogenic endocrine-disrupting chemical actions in the brain using in vivo somatic gene transfer

Author: Caroline Alliot, Vance L. Trudeau, Sébastien Le Mével, Natacha Gallant, Farzad Pakdel, Nathalie Turque, Laurent Coen, Barbara A. Demeneix, Centre for Advanced Research in Environmental Genomics, University of Ottawa [Ottawa], Evolution des régulations endocriniennes (ERE), Muséum national d'Histoire naturelle (MNHN)-Centre National de la Recherche Scientifique (CNRS), Interactions cellulaires et moléculaires (ICM), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes 1 (UR1), and Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Somatic cell, Health, Toxicology and Mutagenesis, bisphenol A, Cytomegalovirus, 010501 environmental sciences, MESH: Goldfish, 01 natural sciences, MESH: Estrogens, Xenopus laevis, Gene expression, MESH: Animals, Luciferases, 0303 health sciences, Gene Transfer Techniques, Transfection, Articles, MESH: Water Pollutants, Chemical, ethinylestradiol, Biological Assay, hormones, hormone substitutes, and hormone antagonists, MESH: Thymidine Kinase, medicine.medical_specialty, MESH: Cytomegalovirus, brain, somatic gene transfer, Endocrine System, MESH: Gene Transfer Techniques, Biology, MESH: Biological Assay, Thymidine Kinase, 03 medical and health sciences, Vitellogenin, MESH: Gene Expression Profiling, MESH: Brain, In vivo, MESH: Xenopus laevis, Internal medicine, Goldfish, medicine, Animals, Luciferase, MESH: Endocrine System, 030304 developmental biology, 0105 earth and related environmental sciences, Hormone response element, Gene Expression Profiling, Research, MESH: Transfection, Public Health, Environmental and Occupational Health, Estrogens, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, estrogen response element, Gene expression profiling, Endocrinology, biology.protein, MESH: Luciferases, Water Pollutants, Chemical
Abstract: International audience; Estrogenic endocrine-disrupting chemicals abnormally stimulate vitellogenin gene expression and production in the liver of many male aquatic vertebrates. However, very few studies demonstrate the effects of estrogenic pollutants on brain function. We have used polyethylenimine-mediated in vivo somatic gene transfer to introduce an estrogen response element-thymidine kinase-luciferase (ERE-TK-LUC) construct into the brain. To determine if waterborne estrogenic chemicals modulate gene transcription in the brain, we injected the estrogen-sensitive construct into the brains of Nieuwkoop-Faber stage 54 Xenopus laevis tadpoles. Both ethinylestradiol (EE2; p < 0.002) and bisphenol A (BPA; p < 0.03) increased luciferase activity by 1.9- and 1.5-fold, respectively. In contrast, low physiologic levels of 17ss-estradiol had no effect (p > 0.05). The mixed antagonist/agonist tamoxifen was estrogenic in vivo and increased (p < 0.003) luciferase activity in the tadpole brain by 2.3-fold. There have been no previous reports of somatic gene transfer to the fish brain; therefore, it was necessary to optimize injection and transfection conditions for the adult goldfish (Carassius auratus). Following third brain ventricle injection of cytomegalovirus (CMV)-green fluorescent protein or CMV-LUC gene constructs, we established that cells in the telencephalon and optic tectum are transfected. Optimal transfections were achieved with 1 microg DNA complexed with 18 nmol 22 kDa polyethylenimine 4 days after brain injections. Exposure to EE2 increased brain luciferase activity by 2-fold in males (p < 0.05) but not in females. Activation of an ERE-dependent luciferase reporter gene in both tadpole and fish indicates that waterborne estrogens can directly modulate transcription of estrogen-responsive genes in the brain. We provide a method adaptable to aquatic organisms to study the direct regulation of estrogen-responsive genes in vivo.
Published: 2005
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30. Specificity of T cells in synovial fluid: high frequencies of CD8(+) T cells that are specific for certain viral epitopes

Author: Tan, LC, Mowat, AG, Fazou, C, Rostron, T, Roskell, H, Dunbar, PR, Tournay, C, Romagné, F, Peyrat, MA, Houssaint, E, Bonneville, M, Rickinson, AB, Mcmichael, AJ, Callan, MF, The Weatherall Institute of Molecular Medicine, University of Oxford, Nuffield Orthopaedic Centre, Immunotech SA, Immunointervention dans les allo et xénotransplantations, Université de Nantes (UN)-IFR26-Institut National de la Santé et de la Recherche Médicale (INSERM)-ITUN, School of Cancer Sciences, University of Birmingham [Birmingham], and Autard, Delphine
Subjects: rheumatoid arthritis, Epstein-Barr Virus Infections, Herpesvirus 4, Human, MESH: Cytomegalovirus, human leucocyte antigen peptide tetrameric complex, MESH: Lymphocyte Count, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Antigens, Surface, MESH: Influenza A virus, Cytomegalovirus, Epitopes, T-Lymphocyte, MESH: Arthritis, CD8-Positive T-Lymphocytes, MESH: Phenotype, MESH: Epitopes, T-Lymphocyte, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Synovial Fluid, Humans, Lymphocyte Count, Antigens, Viral, [SDV.MHEP.RSOA] Life Sciences [q-bio]/Human health and pathology/Rhumatology and musculoskeletal system, MESH: Humans, MESH: Epstein-Barr Virus Infections, Arthritis, MESH: Chronic Disease, Epstein-Barr virus lytic cycle, MESH: Herpesvirus 4, Human, MESH: CD8-Positive T-Lymphocytes, Phenotype, [SDV.MHEP.RSOA]Life Sciences [q-bio]/Human health and pathology/Rhumatology and musculoskeletal system, Influenza A virus, viral immunity, Antigens, Surface, Chronic Disease, MESH: Research Supp, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, [SDV.IMM]Life Sciences [q-bio]/Immunology, Primary Research, CD8+ T cell, MESH: Antigens, Viral
Abstract: CD8+ T cells dominate the lymphocyte population in synovial fluid in chronic inflammatory arthritis. It is known that these CD8+ T cells are often clonally or oligoclonally expanded, but their specificity and their relevance to the pathogenesis of joint disease has remained unclear. We found that as many as 15.5% of synovial CD8+ T cells may be specific for a single epitope from an Epstein-Barr virus lytic cycle protein. The virus-specific T cells within the joint showed increased expression of markers of activation and differentiation compared with those in the periphery, and retained their functional capacity to secrete proinflammatory cytokines on stimulation. These activated, virus-specific CD8+ T cells could therefore interact with synoviocytes, either by cell-cell contact or by a cytokine network, and play a 'bystander' role in the maintenance of inflammation in patients with arthritis., Introduction: Epstein-Barr virus (EBV) is transmitted orally, replicates in the oropharynx and establishes life-long latency in human B lymphocytes. T-cell responses to latent and lytic/replicative cycle proteins are readily detectable in peripheral blood from healthy EBV-seropositive individuals. EBV has also been detected within synovial tissue, and T-cell responses to EBV lytic proteins have been reported in synovial fluid from a patient with rheumatoid arthritis (RA). This raises the question regarding whether T cells specific for certain viruses might be present at high frequencies within synovial fluid and whether such T cells might be activated or able to secrete cytokines. If so, they might play a 'bystander' role in the pathogenesis of inflammatory joint disease. Objectives: To quantify and characterize T cells that are specific for epitopes from EBV, cytomegalovirus (CMV) and influenza in peripheral blood and synovial fluid from patients with arthritis. Methods: Peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) were obtained from patients with inflammatory arthritis (including those with RA, osteoarthritis, psoriatic arthritis and reactive arthritis). Samples from human leucocyte antigen (HLA)-A2-positive donors were stained with fluorescent-labelled tetramers of HLA-A2 complexed with the GLCTLVAML peptide epitope from the EBV lytic cycle protein BMLF1, the GILGFVFTL peptide epitope from the influenza A matrix protein, or the NLVPMVATV epitope from the CMV pp65 protein. Samples from HLA-B8-positive donors were stained with fluorescent-labelled tetramers of HLA-B8 complexed with the RAKFKQLL peptide epitope from the EBV lytic protein BZLF1 or the FLRGRAYGL peptide epitope from the EBV latent protein EBNA3A. All samples were costained with an antibody specific for CD8. CD4+ T cells were not analyzed. Selected samples were costained with antibodies specific for cell-surface glycoproteins, in order to determine the phenotype of the T cells within the joint and the periphery. Functional assays to detect release of IFN-γ or tumour necrosis factor (TNF)-α were also performed on some samples. Results: The first group of 15 patients included 10 patients with RA, one patient with reactive arthritis, one patient with psoriatic arthritis and three patients with osteoarthritis. Of these, 11 were HLA-A2 positive and five were HLA-B8 positive. We used HLA-peptide tetrameric complexes to analyze the frequency of EBV-specific T cells in PBMCs and SFMCs (Figs 1 and 2). Clear enrichment of CD8+ T cells specific for epitopes from the EBV lytic cycle proteins was seen within synovial fluid from almost all donors studied, including patients with psoriatic arthritis and osteoarthritis and those with RA. In donor RhA6, 9.5% of CD8+ SFMCs were specific for the HLA-A2 restricted GLCTLVAML epitope, compared with 0.5% of CD8+ PBMCs. Likewise in a donor with osteoarthritis (NR4), 15.5% of CD8+ SFMCs were specific for the HLA-B8-restricted RAKFKQLL epitope, compared with 0.4% of CD8+ PBMCs. In contrast, we did not find enrichment of T cells specific for the HLA-B8-restricted FLRGRAYGL epitope (from the latent protein EBNA3A) within SFMCs compared with PBMCs in any donors. In selected individuals we performed ELISpot assays to detect IFN-γ secreted by SFMCs and PBMCs after a short incubation in vitro with peptide epitopes from EBV lytic proteins. These assays confirmed enrichment of T cells specific for epitopes from EBV lytic proteins within synovial fluid and showed that subpopulations of these cells were able to secrete proinflammatory cytokines after short-term stimulation. We used a HLA-A2/GILGFVFTL tetramer to stain PBMCs and SFMCs from six HLA-A2-positive patients. The proportion of T cells specific for this influenza epitope was low (
Published: 2000

31. Human cytomegalovirus (HCMV) late-mRNA detection in peripheral blood of AIDS patients: diagnostic value for HCMV disease compared with those of viral culture and HCMV DNA detection

Author: J M Salord, Christos Chouaid, Marie-Caroline Meyohas, C Duvivier, Jean-Claude Petit, O Picard, Joël Gozlan, Epidémiologie des maladies infectieuses et modélisation (ESIM), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Gozlan J, JM Salord, C Chouaïd, Duvivier C, O Picard, Meyohas MC, JC Petit ., and Annesi-Maesano, Isabella
Subjects: Male, Human cytomegalovirus, viruses, Cytomegalovirus, MESH: Base Sequence, medicine.disease_cause, Polymerase Chain Reaction, law.invention, law, Polymerase chain reaction, Viral culture, MESH: AIDS-Related Opportunistic Infections, Evaluation Studies as Topic, MESH: RNA, Viral, Cytomegalovirus Infections, MESH: Evaluation Studies as Topic, RNA, Viral, Female, Viral disease, Research Article, Adult, Microbiology (medical), MESH: Cytomegalovirus, Virus Cultivation, Molecular Sequence Data, Biology, Sensitivity and Specificity, Virus, Herpesviridae, MESH: Virus Cultivation, medicine, Humans, RNA, Messenger, Viremia, MESH: Viremia, MESH: RNA, Messenger, MESH: Molecular Sequence Data, MESH: Humans, AIDS-Related Opportunistic Infections, Base Sequence, MESH: Adult, MESH: Polymerase Chain Reaction, MESH: Cytomegalovirus Infections, medicine.disease, Virology, Reverse transcriptase, MESH: Sensitivity and Specificity, MESH: Male, MESH: DNA, Viral, Viral replication, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, DNA, Viral, Immunology, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, MESH: Female
Abstract: International audience; A reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect a human cytomegalovirus (HCMV) late mRNA in peripheral blood leukocytes (PBL) of 102 human immunodeficiency virus-infected individuals. The clinical value of this new technique for the diagnosis of acute HCMV disease was evaluated in comparison with viral culture and direct amplification of viral DNA (PCR). The sensitivity of the RT-PCR was slightly lower than that of the two other methods, but its specificity was 94%, compared to 55 and 32% for culture and PCR, respectively. Transcription of this late mRNA is linked to viral replication, and its detection in PBL confirms that these cells can support a complete viral cycle. The relationship between complete replicative cycles and HCMV disease makes RT-PCR a useful clinical tool.
Published: 1993

32. New functional domains of human cytomegalovirus pUL89 predicted by sequence analysis and three-dimensional modelling of the catalytic site DEXDc

Author: Serge Bouaziz, Sophie Alain, François Denis, Marie-Christine Mazeron, Stéphanie Stuppfler, Anthony Couvreux, Gaël Champier, Sébastien Hantz, Biologie moléculaire et cellulaire des microorganismes (EA3175), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Bactériologie, Virologie, Hygiène [CHU Limoges], CHU Limoges, Hôpital Lariboisière, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-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)-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)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'informatique Fondamentale de Marseille - UMR 6166 (LIF), Université de la Méditerranée - Aix-Marseille 2-Université de Provence - Aix-Marseille 1-Centre National de la Recherche Scientifique (CNRS), Université Paris Diderot - Paris 7 (UPD7)-Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-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), 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)-Institut de Chimie du CNRS (INC), Bucek, Ingrid, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Lariboisière-Université Paris Diderot - Paris 7 (UPD7), Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL (ENSCP)-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)-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)-Centre National de la Recherche Scientifique (CNRS), Université de Limoges (UNILIM), and Bouaziz, Serge
Subjects: Models, Molecular, Human cytomegalovirus, Protein Conformation, viruses, Cytomegalovirus, MESH: Catalytic Domain, MESH: Amino Acid Sequence, MESH: Drug Design, Virus Replication, Conserved sequence, chemistry.chemical_compound, MESH: Protein Structure, Tertiary, MESH: Protein Conformation, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Catalytic Domain, Pharmacology (medical), Peptide sequence, Conserved Sequence, Adenosine Triphosphatases, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Zinc finger, Genetics, MESH: Conserved Sequence, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], virus diseases, Zinc Fingers, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], Infectious Diseases, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Models, Molecular, MESH: Antiviral Agents, MESH: Cytomegalovirus, [SDV.BBM.BS] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Sequence analysis, Molecular Sequence Data, MESH: Sequence Alignment, Sequence alignment, Biology, Antiviral Agents, Viral Proteins, DNA Packaging, MESH: Polymorphism, Genetic, medicine, MESH: Adenosine Triphosphatases, Humans, MESH: Zinc Fingers, Amino Acid Sequence, MESH: Endodeoxyribonucleases, Binding site, MESH: DNA Packaging, Pharmacology, Endodeoxyribonucleases, Polymorphism, Genetic, MESH: Humans, MESH: Molecular Sequence Data, MESH: Virus Replication, medicine.disease, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, MESH: Viral Proteins, Protein Structure, Tertiary, MESH: DNA, Viral, chemistry, Drug Design, DNA, Viral, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Sequence Alignment, DNA
Abstract: Introduction Benzimidazole d-ribonucleosides inhibit DNA packaging during human cytomegalovirus (HCMV) replication. Although they have been shown to target pUL56 and pUL89, the large and small subunits of the HCMV terminase respectively, their mechanism of action is not yet fully understood. Methods and results To better understand HCMV DNA maturation and the mechanism of action of benzimidazole derivatives, we studied the HCMV pUL89 protein by a genetic approach combined with primary structure analysis. The pUL89 sequence analysis of 25 HCMV strains and counterparts among herpesviruses allowed identification of 12 conserved regions. We also built a three-dimensional model of the pUL89 ATPasic catalytic site, including ATPase motor motifs I, II and III, that may facilitate the development of future antiviral drugs active against HCMV. Finally, we identified several putative functional domains in pUL89, such as pUL89 zinc finger (pUL89-ZF), DNA cutting sites and portal binding sites, that are probably involved in CMV DNA cleavage and packaging.

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32 results on '"MESH: Cytomegalovirus"'

1. Sémiologie et pronostic de l’infection congénitale à cytomégalovirus après primo-infection maternelle du second trimestre de la grossesse

2. Repertoire characterization and validation of gB-specific human IgGs directly cloned from humanized mice vaccinated with dendritic cells and protected against HCMV

3. Pronostic du fœtus infecté par le cytomégalovirus après primo-infection maternelle du premier trimestre

4. CMV et grossesse : connaissances et pratiques des professionnels de santé

5. Conserved domains and structure prediction of human cytomegalovirus UL27 protein

6. Étude des facteurs prédictifs de séquelles à 2 ans en lien avec une infection congénitale à Cytomégalovirus

7. Aminobisphosphonates Synergize with Human Cytomegalovirus To Activate the Antiviral Activity of Vγ9Vδ2 Cells

8. Therapeutic drug monitoring of ganciclovir in the management of cytomegalovirus : should we reassess the therapeutic range ? Results of a preliminary study conducted at the University Hospital of Rennes

9. Development and validation of a non-radioactive DNA polymerase assay for studying cytomegalovirus resistance to foscarnet

10. Retrograde transport is not required for cytosolic translocation of the B-subunit of Shiga toxin

11. A Novel Mutation in the Ul54 Gene of Human Cytomegalovirus Isolates that Confers Resistance to Foscarnet

12. Human Cytomegalovirus Binding to DC-SIGN Is Required for Dendritic Cell Infection and Target Cell trans-Infection

13. Success and failure of artesunate treatment in five transplant recipients with disease caused by drug-resistant cytomegalovirus

14. Human herpes virus co-infection is associated with mortality in HIV-negative patients with Pneumocystis jirovecii pneumonia

15. Insight into the structure of the pUL89 C-terminal domain of the human cytomegalovirus terminase complex

16. Structural bases for the affinity-driven selection of a public TCR against a dominant human cytomegalovirus epitope

17. Bcl-3-regulated transcription from major immediate-early promoter of human cytomegalovirus in monocyte-derived macrophages

18. Cytomegalovirus microRNAs

19. Direct genotyping of cytomegalovirus envelope glycoproteins from toddler's saliva samples

20. Rapid determination of antiviral drug susceptibility of human cytomegalovirus by real-time PCR

21. Early immune response against retrovirally transduced herpes simplex virus thymidine kinase-expressing gene-modified T cells coinfused with a T cell-depleted marrow graft: an altered immune response?

22. Replication and packaging of an infectious bursal disease virus segment A-derived minigenome

23. Putative functional domains of human cytomegalovirus pUL56 involved in dimerization and benzimidazole D-ribonucleoside activity

24. Maintenance of HIV-specific central and effector memory CD4 and CD8 T cells requires antigen persistence

25. [Cytomegalovirus and cryptogenic inflammatory bowel disease]

26. Monitoring human cytomegalovirus (HCMV) in HCMV-seropositive orthotopic liver-transplant recipients by means of quantitative real-time polymerase chain reaction

27. Multiplex real-time PCR assay for simultaneous quantitation of human cytomegalovirus and herpesvirus-6 in polymorphonuclear and mononuclear cells of transplant recipients

28. Phenotypic characterisation of cytomegalovirus DNA polymerase: a method to study cytomegalovirus isolates resistant to foscarnet

29. Assessment of estrogenic endocrine-disrupting chemical actions in the brain using in vivo somatic gene transfer

30. Specificity of T cells in synovial fluid: high frequencies of CD8(+) T cells that are specific for certain viral epitopes

31. Human cytomegalovirus (HCMV) late-mRNA detection in peripheral blood of AIDS patients: diagnostic value for HCMV disease compared with those of viral culture and HCMV DNA detection

32. New functional domains of human cytomegalovirus pUL89 predicted by sequence analysis and three-dimensional modelling of the catalytic site DEXDc

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32 results on '"MESH: Cytomegalovirus"'

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