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45 results on '"Neuro-Immunologie Virale"'

1. Predominant role of microglia in brain iron retention in Sanfilippo syndrome, a pediatric neurodegenerative disease

Author

Laurent Puy, Cathy Gomila, Jérôme Ausseil, Agnès Boullier, Zoubida Karim, Stéphanie Trudel, Christelle Lony, Sandrine Vitry, Thibaud Lefebvre, Walaa Darwiche, Vincent Puy, Université de Picardie Jules Verne (UPJV), North Hospital, Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Inserm, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre hospitalier universitaire d'Amiens (CHU Amiens-Picardie), CHU Amiens-Picardie, Mécanismes physiologiques et conséquences des calcifications cardiovasculaires: rôle des remodelages cardiovasculaires et osseux, Université de Picardie Jules Verne (UPJV)-Institut National de la Santé et de la Recherche Médicale (INSERM), HEMATIM - Hématopoïèse et immunologie - UR UPJV 4666 (HEMATIM), CHU Amiens-Picardie-Université de Picardie Jules Verne (UPJV)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale - Viral Neuro-immunology, This work was funded by the Vaincre les Maladies Lysosomales (VML) charity., We thank Professor Elizabeth Neufeld for providing the MPSIIIB mice, and Dr. Bénédicte Heron (APHP, Hôpital Trousseau, Paris, France) for collecting urine samples from patients. We are grateful to Camille Rottier (CHU Amiens Picardie, Amiens, France), Dr. Julie Bruyère (Institut des Neurosciences, Grenoble) and Nathalie Dessendier (Centre Français des Porphyries ‐ Laboratoire de Biochimie, Hôpital Louis Mourier, Colombes, France) for excellent technical assistance. We thank the staff of ICAP plateform, Dr. Paulo Marcelo and Mrs. Luciane Zabijak, for assistance with confocal microscopy experiments (ICAP plateform, Université de Picardie Jules Verne, Amiens, France)., Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Mécanismes physiopathologiques et conséquences des calcifications vasculaires - UR UPJV 7517 (MP3CV), and Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie

Subjects
0301 basic medicine, Sanfilippo syndrome, MESH: Neurons, microglia, MESH: Mice, Knockout, [SDV.MHEP.UN]Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology, neuroinflammation, MESH: Neurodegenerative Diseases, Mucopolysaccharidosis III, chemistry.chemical_compound, iron, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Animals, MESH: Mucopolysaccharidosis III, Mice, Knockout, Neurons, MESH: Iron, Microglia, Brain, Neurodegenerative Diseases, [SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology, Heparan sulfate, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, MESH: Microglia, medicine.anatomical_structure, Neurology, Cerebral cortex, medicine.medical_specialty, Mucopolysaccharidosis type III, Biology, MESH: Brain, 03 medical and health sciences, Cellular and Molecular Neuroscience, Hepcidin, Internal medicine, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], medicine, Animals, Neuroinflammation, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, medicine.disease, MESH: Astrocytes, 030104 developmental biology, Endocrinology, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, chemistry, Astrocytes, TLR4, biology.protein, hepcidin, 030217 neurology & neurosurgery
Abstract

International audience; Neuroinflammation and iron accumulation are hallmarks of a variety of adult neurodegenerative diseases. In Sanfilippo syndrome (mucopolysaccharidosis type III, MPSIII, a pediatric neurodegenerative disease that shares some features with adult neurodegenerative diseases), the progressive accumulation of heparan sulfate oligosaccharides (HSOs) induces microglia and astrocytes to produce pro-inflammatory cytokines leading to severe neuroinflammation. The objectives of the present study were (1) to measure the local iron concentration and to assess iron metabolism in the brain of a MPSIIIB murine model and (2) to identify the brain cells involved in this accumulation. We found that iron accumulation in MPSIIIB mice primarily affected the cerebral cortex where hepcidin levels were higher than in wild-type mice, and increased with aging. This increase was correlated with low expression of ferroportin 1 (FPN1), and thus brain iron retention. Moreover, we showed in vitro that HSOs are directly responsible for the production of hepcidin and the relative decrease in FPN1 expression when added to cultures of microglia and, to a lesser extent, to cultures of astrocytes. In contrast, no significant differences were observed in neurons. Hepcidin induction results from activation of the TLR4 pathway and STAT3 signaling, and leads to iron retention within microglia. Our results show that microglia have a key role in cerebral hepcidin overexpression and thus in the brain iron accumulation observed in the MPSIIIB model.

Published
2018

2. Structure-based optimization of a PDZ binding motif within a viral peptide stimulates neurite outgrowth

Author

Cynthia Lefebvre-Omar, Alain Chaffotte, Florent Delhommel, Christophe Prehaud, Elouan Terrien, Sandrine Vitry, Renaud Vincentelli, Clara Bernard, Zakir Khan, Kevin Ricquier, Delphine Bohl, Monique Lafon, Nicolas Wolff, Henri Buc, Juan Ramirez, Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Résonance Magnétique Nucléaire des Biomolécules, Institut du Cerveau = Paris Brain Institute (ICM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Architecture et fonction des macromolécules biologiques (AFMB), Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), This work was supported by the following grants: ANR (Agence Nationale de la Recherche), BSV8- 0008-01 MAST-2-PTEN. AFM (Association Française contre les Myopathies) Characterization of pathological defects in motor neurons derived from patients Amyotrophic Lateral Sclerosis (ALS). Grand Prix de l’Académie des Sciences/Fondation Générale de Santé, and Incentive grant from Institut Carnot -Pasteur Maladies Infectieuses Unraveling the neuroprotective functions and optimization of new neuroprotective compound . The research leading to the results on IP SC has received funding from the programs Investissements d’Avenir ANR -10- IAIHU -06 and ANR -11- INBS -0011- NeurATRIS : Translational Research Infrastructure for Biotherapies in Neurosciences. ET and FD are recipients of PhD fellowships from the Ministère de l’ Enseignement Supérieur et de la Recherche. CL-O was supported by a PhD fellowship from Revive (ANR -10 -LABX -73). We thank Sylviane Hoos (Biophysics Core, Institut Pasteur) for ITC expertise and the team of the Technology Core of the Center for Translational Science (CRT), Institut Pasteur, for their help our with the Incucyte Technology., ANR-13-BSV8-0008,MAST-2-PTEN,Etude structurale et fonctionnelle du complexe MAST2/PTEN, impliqué dans le contrôle de la survie neuronale.(2013), ANR-11-INBS-0011,NeurATRIS,Infrastructure de Recherche Translationnelle pour les Biothérapies en Neurosciences(2011), ANR-10-LABX-0073,REVIVE,Stem Cells in Regenerative Biology and Medicine(2010), Binet, Caroline, Blanc 2013 - Etude structurale et fonctionnelle du complexe MAST2/PTEN, impliqué dans le contrôle de la survie neuronale. - - MAST-2-PTEN2013 - ANR-13-BSV8-0008 - Blanc 2013 - VALID, Infrastructures - Infrastructure de Recherche Translationnelle pour les Biothérapies en Neurosciences - - NeurATRIS2011 - ANR-11-INBS-0011 - INBS - VALID, Laboratoires d'excellence - Stem Cells in Regenerative Biology and Medicine - - REVIVE2010 - ANR-10-LABX-0073 - LABX - VALID, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut du Cerveau et de la Moëlle Epinière = Brain and Spine Institute (ICM), Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [APHP]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], ANR: 11-INBS-0011,NeurATRIS,Infrastructure de Recherche Translationnelle pour les Biothérapies en Neurosciences(2011), ANR-10-LABX-0073/10-LABX-0073,REVIVE,Stem Cells in Regenerative Biology and Medicine(2010), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut National de la Santé et de la Recherche Médicale (INSERM)-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) (AP-HP)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA)

Subjects
0301 basic medicine, viral protein, neurite outgrowth, Neurite, Viral protein, structure function analysis, Nuclear Magnetic Resonance, Induced Pluripotent Stem Cells, Neuronal Outgrowth, PDZ domain, PDZ Domains, Peptide, Serine threonine protein kinase, Protein Serine-Threonine Kinases, medicine.disease_cause, Microtubules, Biochemistry, Protein–protein interaction, protein-protein interaction, Structure-Activity Relationship, Viral Proteins, 03 medical and health sciences, [SCCO]Cognitive science, Neurites, medicine, Humans, Protein Interaction Domains and Motifs, neurodegenerative diseases, Molecular Biology, Neurons, chemistry.chemical_classification, 030102 biochemistry & molecular biology, Kinase, [SCCO.NEUR]Cognitive science/Neuroscience, [SCCO.NEUR] Cognitive science/Neuroscience, Cell Biology, [SCCO] Cognitive science, serine/threonine protein kinase, Cell biology, 030104 developmental biology, chemistry, Rabies virus, Cytoplasm, Central Nervous System Stimulants, Interfering peptide, Peptides, therapeutical drug candidate
Abstract

International audience; Protection of neuronal homeostasis is a major goal in the management of neurodegenerative diseases. Microtubule-associated Ser/Thre kinase 2 (MAST2), inhibits neurite outgrowth, and its inhibition therefore represents a potential therapeutic strategy. We previously reported that a viral protein (G-protein from rabies virus) capable of interfering with protein-protein interactions between the PDZ domain of MAST2 and the C-terminal moieties of its cellular partners counteracts MAST2-mediated suppression of neurite outgrowth. Here, we designed peptides derived from the native viral protein to increase the affinity of these peptides for the MAST2 PDZ domain. Our strategy involved modifying the length and flexibility of the non-interacting sequence linking the two subsites anchoring the peptide to the PDZ domain. Three peptides, Neurovita1 (NV1), NV2, and NV3, were selected, and we found that they all had increased affinities for the MAST2 PDZ domain, with Kd values decreasing from 1300 to 60 nM, while target selectivity was maintained. A parallel biological assay evaluating neurite extension and branching in cell cultures revealed that the NV peptides gradually improved neural activity, with the efficacies of these peptides for stimulating neurite outgrowth mirroring their affinities for MAST2-PDZ. We also show that NVs can be delivered into the cytoplasm of neurons as a gene or peptide. In summary, our findings indicate that virus-derived peptides targeted to MAST2-PDZ stimulate neurite outgrowth in several neuron types, opening up promising avenues for potentially using NVs in the management of neurodegenerative diseases.

Published
2019

3. A Human Blood-Brain Interface Model to Study Barrier Crossings by Pathogens or Medicines and Their Interactions with the Brain

Author

Anaelle da Costa, Monique Lafon, Florian Bakoa, Pierre Lafaye, Philippe V. Afonso, Pierre-Emmanuel Ceccaldi, Christophe Prehaud, Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Epidémiologie et Physiopathologie des Virus Oncogènes (EPVO (UMR_3569 / U-Pasteur_3)), Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)-Institut Pasteur [Paris], Ingénierie des Anticorps (plate-forme) - Antibody Engineering (Platform), This study was supported by internal grants from Institut Pasteur including an Incitative grant (PTR 435) and by a grant 'Contrat de Soutien à la Recherche' provided by Sanofi Pasteur to Institut Pasteur. A. da Costa was supported by the Sanofi-Pasteur grant and Florian Bakoa is recipient of a PhD grant provided by ANRT (Association Nationale de la Recherche et de la Technologie), We are indebted to Pr Pierre-Olivier Couraud and Dr Florence Miller for helpful discussions., Neuro-Immunologie Virale, Institut Pasteur [Paris]-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Nervous system, Drug, Interface model, General Chemical Engineering, media_common.quotation_subject, Models, Neurological, 030231 tropical medicine, Neuroprotection, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, [SCCO]Cognitive science, 0302 clinical medicine, Parenchyma, Animals, Humans, Medicine, Animal testing, Cells, Cultured, media_common, Neurons, General Immunology and Microbiology, business.industry, General Neuroscience, [SCCO.NEUR]Cognitive science/Neuroscience, Brain, Endothelial Cells, Human brain, 3. Good health, Neuroprotective Agents, 030104 developmental biology, medicine.anatomical_structure, Drug development, nervous system, Blood-Brain Barrier, Astrocytes, cardiovascular system, business, Neuroscience
Abstract

International audience; The early screening of nervous system medicines on a pertinent and reliable in cellulo BBB model for their penetration and their interaction with the barrier and the brain parenchyma is still an unmet need. To fill this gap, we designed a 2D in cellulo model, the BBB-Minibrain, by combining a polyester porous membrane culture insert human BBB model with a Minibrain formed by a tri-culture of human brain cells (neurons, astrocytes and microglial cells). The BBB-Minibrain allowed us to test the transport of a neuroprotective drug candidate (e.g., Neurovita), through the BBB, to determine the specific targeting of this molecule to neurons and to show that the neuroprotective property of the drug was preserved after the drug had crossed the BBB. We have also demonstrated that BBB-Minibrain constitutes an interesting model to detect the passage of virus particles across the endothelial cells barrier and to monitor the infection of the Minibrain by neuroinvasive virus particles. The BBB-Minibrain is a reliable system, easy to handle for researcher trained in cell culture technology and predictive of the brain cells phenotypes after treatment or insult. The interest of such in cellulo testing would be twofold: introducing derisking steps early in the drug development on the one hand and reducing the use of animal testing on the other hand.

Published
2019

4. Botulinum Neurotoxin Type B Uses a Distinct Entry Pathway Mediated By Cdc42 Into Intestinal Cells Versus Neuronal Cells

Author

Marie Voillequin, Michel R. Popoff, Christelle Mazuet, Sandrine Vitry, Alain Vandewalle, Christian Lévêque, Carolina Varela Chavez, Chloé Connan, Bactéries anaérobies et Toxines, Institut Pasteur [Paris], Unité de Neurobiologie des canaux Ioniques et de la Synapse (UNIS - Inserm U1072), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale, and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7)

Subjects
0301 basic medicine, Cell type, Endosome, Immunology, intestinal cells, Biology, medicine.disease_cause, Endocytosis, Microbiology, neuronal cells, 03 medical and health sciences, Mice, Intestinal mucosa, Virology, medicine, Clostridium botulinum, Animals, endocytosis, Botulism, Cdc42, Botulinum Toxins, Type A, cdc42 GTP-Binding Protein, Neurons, 030102 biochemistry & molecular biology, Infant Botulism, Synaptotagmin I, botulinum neurotoxin, Epithelial Cells, medicine.disease, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, gangliosides, 3. Good health, Cell biology, 030104 developmental biology, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]
Abstract

Botulinum neurotoxins (BoNTs) are responsible for severe flaccid paralysis by inhibiting the release of acetylcholine at the neuromuscular junctions. BoNT type B (BoNT/B) most often induces mild forms of botulism with predominant dysautonomic symptoms. In food borne botulism and botulism by intestinal colonisation such as infant botulism, which are the most frequent naturally acquired forms of botulism, the digestive tract is the main entry route of BoNTs into the organism. We previously showed that BoNT/B translocates through mouse intestinal barrier by an endocytosis-dependent mechanism and subsequently targets neuronal cells, mainly cholinergic neurons, in the intestinal mucosa and musculosa. Here, we investigated the entry pathway of BoNT/B using fluorescent C-terminal domain of the heavy chain (HcB), which is involved in the binding to specific receptor(s) and entry process into target cells. While the combination of gangliosides GD1a /GD1b /GT1b and synaptotagmin I and to a greater extent synaptotagmin II constitutes the functional HcB receptor on NG108-15 neuronal cells, HcB only uses the gangliosides GD1a /GD1b /GT1b to efficiently bind to m-ICcl2 intestinal cells. HcB enters both cell types by a dynamin-dependent endocytosis, which is efficiently prevented by Dynasore, a dynamin inhibitor, and reaches a common early endosomal compartment labeled by early endosome antigen (EEA1). In contrast to neuronal cells, HcB uses a Cdc42-dependent pathway to enter intestinal cells. Then, HcB is transported to late endosomes in neuronal cells, whereas it exploits a nonacidified pathway from apical to basal lateral side of m-ICcl2 cells supporting a transcytotic route in epithelial intestinal cells.

Published
2017

5. Role of the glycoprotein thorns in anxious effects of rabies virus: Evidence from an animal study

Author

Soheil, Ghassemi, Hamid Gholami, Pourbadie, Christophe, Prehaud, Monique, Lafon, Mohammad, Sayyah, Institut Pasteur d'Iran, Réseau International des Instituts Pasteur (RIIP), Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and This work is part of the Ph.D. dissertation of Soheil Ghassemi ( PP. 9349) and supported by Pasteur Institute of Iran.

Subjects
Hypothalamo-Hypophyseal System, Rabies, [SDV]Life Sciences [q-bio], General Neuroscience, Pituitary-Adrenal System, Anxiety, Rabies envelope, Rats, Rabies virus, PBM, Animals, Humans, Corticosterone, Elevated plus maze, Glycoproteins
Abstract

International audience; Rabies is a lethal infectious disease caused by rabies virus (RABV). Fear and anxiety are the distinguished symptoms in rabies patients. Fusion of RABV envelope glycoprotein (RVG) to host cell membrane initiates rabies pathogenesis via interacting with PDZ domain of signaling proteins. We assessed the anxiety-like behaviors, and hypothalamic-pituitary-adrenal axis (HPA) response to RVG infection. Contribution of PDZ binding motif (PBM) of RVG to the observed effects was also examined using a mutant form of RVG, ΔRVG, with deleted last four amino acids at PBM C-terminus. Lentiviral vectors containing RVG and/or ΔRVG genes were injected into the rat brain areas involved in anxiety including hypothalamus, dorsal hippocampus, and amygdala. RVG/ΔRVG neural expression was examined by fluorescent microscopy. Anxiety-like behaviors were assessed by elevated plus maze (EPM) and open field (OF) tasks. HPA response was evaluated via measuring corticosterone serum level by ELISA technique. RVG/ΔRVG were successfully expressed in neurons of the injected areas. RVG, but not ΔRVG, infection of hypothalamus and amygdala increased the time spent in EPM open arms, and OF total distance moved and velocity. RVG, but not ΔRVG, infection of hypothalamus and dorsal hippocampus increased corticosterone level. The anxiety-like behaviors and exploratory/locomotor activities of rats with RVG infection in hypothalamus, and amygdala are mediated by PBM of RVG. The HPA response to RVG infection of hypothalamus and dorsal hippocampus is dependent to PBM of RVG. Triggering anxiety-related signaling by PBM of RVG seems to be one of the mechanisms involved in anxiety behaviors seen in patients with rabies.

Published
2022

6. Lentiviral Expression of Rabies Virus Glycoprotein in the Rat Hippocampus Strengthens Synaptic Plasticity

Author

Kayhan Azadmanesh, Mohammad Sayyah, Monique Lafon, Nima Naderi, Soheil Ghassemi, Alireza Gholami, Hadi Mirzapourdelavar, Christophe Prehaud, Hamid Gholami Pourbadie, Shayan Aliakbari, Tara Asgari, Institut Pasteur d'Iran, Réseau International des Instituts Pasteur (RIIP), Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Shahid Beheshti University of Medical Sciences [Tehran] (SBUMS), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Shahid Beheshti University

Subjects
0301 basic medicine, Plasticity, Long-Term Potentiation, Population, Hippocampus, Hippocampal formation, Biology, medicine.disease_cause, 03 medical and health sciences, Cellular and Molecular Neuroscience, 0302 clinical medicine, Postsynaptic potential, medicine, Animals, Dentate gyrus, education, Glycoproteins, education.field_of_study, Neuronal Plasticity, Rabies virus, Long-term potentiation, Cell Biology, General Medicine, Long term potentiation, Electric Stimulation, Rats, PDZ binding motif, 3. Good health, Cell biology, 030104 developmental biology, Synaptic plasticity, [SDV.IMM]Life Sciences [q-bio]/Immunology, 030217 neurology & neurosurgery
Abstract

“This is a pre-print of an article published in journal of Cellular and Molecular Neurobiology. The final authenticated version is available online at: https://doi.org/10.1007/s10571-020-01032-9"; International audience; Rabies virus (RABV) is a neurotropic virus exclusively infecting neurons in the central nervous system. RABV encodes five proteins. Among them, the viral glycoprotein (RVG) plays a key role in viral entry into neurons and rabies pathogenesis. It was shown that the nature of the C-terminus of the RABV G protein, which possesses a PDZ-binding motif (PBM), modulates the virulence of the RABV strain. The neuronal protein partners recruited by this PBM may alter host cell function. This study was conducted to investigate the effect of RVG on synaptic function in the hippocampal dentate gyrus (DG) of rat. Two μl (108 T.U./ml) of the lentiviral vector containing RVG gene was injected into the DG of rat hippocampus. After 2 weeks, the rat’s brain was cross-sectioned and RVG-expressing cells were detected by fluorescent microscopy. Hippocampal synaptic activity of the infected rats was then examined by recording the local field potentials from DG after stimulation of the perforant pathway. Short-term synaptic plasticity was also assessed by double pulse stimulation. Expression of RVG in DG increased long-term potentiation population spikes (LTP-PS), whereas no facilitation of LTP-PS was found in neurons expressing δRVG (deleted PBM). Furthermore, RVG and δRVG strengthened paired-pulse facilitation. Heterosynaptic long-term depression (LTD) in the DG was significantly blocked in RVG-expressing group compared to the control group. This blockade was dependent to PBM motif as rats expressing δRVG in the DG-expressed LTD comparable to the RVG group. Our data demonstrate that RVG expression facilitates both short- and long-term synaptic plasticity in the DG indicating that it may involve both pre- and postsynaptic mechanisms to alter synaptic function. Further studies are needed to elucidate the underlying mechanisms.

Published
2021

7. Genomic diversity contributes to the neuroinvasiveness of the Yellow fever French neurotropic vaccine

Author

Bakoa, Florian, Préhaud, Christophe, Beauclair, Guillaume, Chazal, Maxime, Mantel, Nathalie, Lafon, Monique, Jouvenet, Nolwenn, Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Sanofi Pasteur [Marcy-l'Étoile, France], Collège doctoral [Sorbonne universités], Sorbonne Université (SU), Signalisation antivirale - Virus sensing and signaling, Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), We thank the Association Nationale de la Recherche et de la Technologie (CIFRE 2017/0314), Sanofi-Pasteur and Institut Pasteur for funding., Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Collège Doctoral

Subjects
Vaccines, Live attenuated vaccines, Virology, [SDV]Life Sciences [q-bio], Neoplasms. Tumors. Oncology. Including cancer and carcinogens, Immunologic diseases. Allergy, RC581-607, Article, RC254-282
Abstract

International audience; Mass vaccination with the live attenuated vaccine YF-17D is the current way to prevent infection with Yellow fever virus (YFV). However, 0.000012-0.00002% of vaccinated patients develop post-vaccination neurological syndrome (YEL-AND). Understanding the factors responsible for neuroinvasion, neurotropism, and neurovirulence of the vaccine is critical for improving its biosafety. The YF-FNV vaccine strain, known to be associated with a higher frequency of YEL-AND (0.3-0.4%) than YF-17D, is an excellent model to study vaccine neuroinvasiveness. We determined that neuroinvasiveness of YF-FNV occured both via infection and passage through human brain endothelial cells. Plaque purification and next generation sequencing (NGS) identified several neuroinvasive variants. Their neuroinvasiveness was not higher than that of YF-FNV. However, rebuilding the YF-FNV population diversity from a set of isolated YF-FNV-N variants restored the original neuroinvasive phenotype of YF-FNV. Therefore, we conclude that viral population diversity is a critical factor for YFV vaccine neuroinvasiveness.

Published
2021

8. Rabies virus glycoprotein enhances spatial memory via the PDZ binding motif

Author

Soheil Ghassemi, Monique Lafon, Christophe Prehaud, Nima Naderi, Alireza Gholami, Tara Asgari, Kayhan Azadmanesh, Mohammad Sayyah, Hamid Gholami Pourbadie, Institut Pasteur d'Iran, Réseau International des Instituts Pasteur (RIIP), Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Shahid Beheshti University of Medical Sciences [Tehran] (SBUMS), Shahid Beheshti University, and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, 0301 basic medicine, Gene Expression, Morris water navigation task, Hippocampus, Hippocampal formation, medicine.disease_cause, CA1, Stereotaxic Techniques, 0302 clinical medicine, Genes, Reporter, Transgenes, Spatial Memory, Neurons, chemistry.chemical_classification, 3. Good health, Cell biology, PDZ binding motif, Neurology, [SDV.IMM]Life Sciences [q-bio]/Immunology, Signal transduction, learning and memory, dorsal hippocampus, Recombinant Fusion Proteins, Genetic Vectors, Green Fluorescent Proteins, PDZ domain, Biology, Viral vector, Viral Proteins, 03 medical and health sciences, Cellular and Molecular Neuroscience, Virology, Avoidance Learning, medicine, Animals, Rats, Wistar, Maze Learning, CA1 Region, Hippocampal, Glycoproteins, Injections, Intraventricular, Lentivirus, Rabies virus, Rats, 030104 developmental biology, chemistry, rabies glycoprotein, Neurology (clinical), Glycoprotein, 030217 neurology & neurosurgery
Abstract

“This is a pre-print of an article published in J. Neurovirol. The final authenticated version is available online at: https://doi.org/10.1007/s13365-021-00972-2”; International audience; Rabies is a life-threatening viral infection of the brain. Rabies virus (RABV) merely infects excitable cells including neurons provoking drastic behaviors including negative emotional memories. RABV glycoprotein (RVG) plays a critical role in RABV pathogenesis. RVG interacts with various cytoplasmic PDZ (PSD-95/Dlg/ZO-1) containing proteins through its PDZ binding motif (PBM). PTZ domains have crucial role in formation and function of signal transduction. Hippocampus is one of the cerebral regions that contains high load of viral antigens. We examined impact of RVG expression in the dorsal hippocampus on aversive as well as spatial learning and memory performance in rats. Two μl of the lentiviral vector (~108 T.U. /ml) encoding RVG or ∆RVG (deleted PBM) genomes was microinjected into the hippocampal CA1. After one week, rat’s brain was cross-sectioned and RVG/∆RVG-expressing neuronal cells were confirmed by fluorescent microscopy. Passive avoidance and spatial learning and memory were assessed in rats by Shuttle box and Morris water maze (MWM). In the shuttle box, both RVG and ∆RVG decreased the time spent in the dark compartment compared to control (p

Published
2021

9. Spatiotemporal analysis of mycolactone distribution in vivo reveals partial diffusion in the central nervous system

Author

Thomas Laval, Sarah Saint-Auret, Ivo G. Boneca, Laurent Boucontet, Caroline Demangel, Jean-Pierre Levraud, Laure Guenin-Macé, Christophe Prehaud, Anaelle da Costa, Emma Colucci-Guyon, Aline Rifflet, Nicolas Blanchard, Macrophages et Développement de l’Immunité, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Biologie et Génétique de la Paroi bactérienne - Biology and Genetics of Bacterial Cell Wall, Laboratoire d'innovation moléculaire et applications (LIMA), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale - Viral Neuro-immunology, Immunobiologie de l'Infection - Immunobiology of Infection, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Diderot, Sorbonne Paris Cité, Paris, France, Université Paris Diderot - Paris 7 (UPD7), This work was supported by the Institut Pasteur 'Microbes & Brain' collaborative project (LGM) and the Fondation Raoul Follereau (CD), core support from Institut Pasteur and INSERM (U1221). This study has received funding from the French Government’s Investissement d’Avenir program, Laboratoire d’Excellence 'Integrative Biology of Emerging Infectious Diseases' (grant n °ANR-10-LABX-62-IBEID) (IGB). AR was support by a post-doctoral fellowship from the Laboratoire d’Excellence 'Integrative Biology of Emerging Infectious Diseases. T.L. was a BioSPC-Université Paris Diderot PhD student, recipient of doctoral fellowships from the Ministère français de l’Enseignement Supérieur, de la Recherche et de l’Innovation (2016–2019) and Fondation pour la Recherche Médicale (FDT201904008040)., We thank the Image Analysis Hub of the Institut Pasteur for help with image analysis. We are grateful to Laurent Marsollier for the malaysian human isolate M. ulcerans 1615 and to Tim Stinear for the bioluminescent strain of M.ulcerans., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
Central Nervous System, Bacterial Diseases, 0301 basic medicine, Buruli ulcer, Life Cycles, [SDV]Life Sciences [q-bio], RC955-962, Nervous System, chemistry.chemical_compound, Larvae, Medical Conditions, 0302 clinical medicine, Animal Cells, Arctic medicine. Tropical medicine, Medicine and Health Sciences, Mycolactone, Buruli Ulcer, Zebrafish, biology, Optical Imaging, Eukaryota, Animal Models, 3. Good health, Cell biology, Actinobacteria, Infectious Diseases, medicine.anatomical_structure, Experimental Organism Systems, Osteichthyes, Blood-Brain Barrier, Larva, Mycobacterium ulcerans, Vertebrates, Physical Sciences, [SDV.IMM]Life Sciences [q-bio]/Immunology, Macrolides, Cellular Types, Anatomy, Public aspects of medicine, RA1-1270, Research Article, Neglected Tropical Diseases, Imaging Techniques, Materials Science, Material Properties, Bacterial Toxins, Central nervous system, Glial Cells, Research and Analysis Methods, Blood–brain barrier, Permeability, Cell Line, 03 medical and health sciences, Model Organisms, Spatio-Temporal Analysis, In vivo, Fluorescence Imaging, medicine, Animals, Humans, Microglial Cells, Bacteria, Organisms, Public Health, Environmental and Occupational Health, Biology and Life Sciences, Endothelial Cells, Cell Biology, Tropical Diseases, biology.organism_classification, medicine.disease, In vitro, Fish, 030104 developmental biology, chemistry, Astrocytes, Animal Studies, Zoology, 030217 neurology & neurosurgery, Developmental Biology
Abstract

Mycobacterium ulcerans, the causative agent of Buruli ulcer (BU) disease, is unique amongst human pathogens in its capacity to produce a lipid toxin called mycolactone. While previous studies have demonstrated that bacterially-released mycolactone diffuses beyond infection foci, the spatiotemporal distribution of mycolactone remained largely unknown. Here, we used the zebrafish model to provide the first global kinetic analysis of mycolactone’s diffusion in vivo, and multicellular co-culture systems to address the critical question of the toxin’s access to the brain. Zebrafish larvae were injected with a fluorescent-derivative of mycolactone to visualize the in vivo diffusion of the toxin from the peripheral circulation. A rapid, body-wide distribution of mycolactone was observed, with selective accumulation in tissues near the injection site and brain, together with an important excretion through the gastro-intestinal tract. Our conclusion that mycolactone reached the central nervous system was reinforced by an in cellulo model of human blood brain barrier and a mouse model of M. ulcerans-infection. Here we show that mycolactone has a broad but heterogenous profile of distribution in vivo. Our investigations in vitro and in vivo support the view that a fraction of bacterially-produced mycolactone gains access to the central nervous system. The relative persistence of mycolactone in the bloodstream suggests that assays of circulating mycolactone are relevant for BU disease monitoring and treatment optimization., Author summary Mycolactone is the major virulence factor of Mycobacterium ulcerans, the human pathogen causing Buruli ulcer (BU) disease. While it is now established that mycolactone is able to diffuse from infected tissues to exert immunomodulatory and analgesic effects at the systemic level, the in vivo spatiotemporal distribution of mycolactone remained largely unknown. Here, using the zebrafish larva, we describe the spatiotemporal distribution of a fluorescent derivative of mycolactone in vivo. We show that fluorescent mycolactone quickly diffuses from the blood circulation into various organs, and accumulates in muscles and brain. Mycolactone’s diffusion into the central nervous system was further confirmed in the mouse model of M. ulcerans infection. We reported previously that mycolactone suppresses the production of inflammatory mediators by primary microglia at nanomolar concentrations in vitro [1]. In the present study, we provide evidence suggesting that bacterially-produced mycolactone is able to reach the brain. While additional in vivo investigations will be required, we can speculate that the amount of mycolactone reaching the brain in the context of M. ulcerans infection is sufficient to modulate inflammation and pain transmission.

Published
2020

10. Broad spectrum compounds targeting early stages of rabies virus (RABV) infection

Author

Sabrina Kali, Saliha Azebi, Corinne Jallet, Juliana Pipoli Da Fonseca, Daniel Gillet, Jean-Christophe Cintrat, Noël Tordo, Thomas Cokelaer, Yu Wu, Julien Barbier, Stratégies antivirales, Institut Pasteur [Paris], Institut Pasteur d'Algérie, Réseau International des Instituts Pasteur (RIIP), Biomics (plateforme technologique), Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), 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), Service de Chimie Bio-Organique et de Marquage (SCBM), Institut Pasteur de Guinée, The following programs contributed to fund the current research, both for material and fellowships: the joint ministerial program of R&D against CBRNE risks and Centre d’Energie Atomique (CEA - P/NRBC n°I1.1.8 'Large Spectrum Antivirals'), the European program ASKLEPIOS « Advanced Studies towards Knowledge on Lyssavirus Encephalitis Pathogenesis Improving Options for Survival » (FP7 - 602825 - Neglected infectious diseases of Central and Eastern Europe). In addition, the project benefited from the core facilities of the « Centre de Ressources et Recherches Technologiques » at the Institut Pasteur for imaging and bioinformatics., European Project: 602825,EC:FP7:HEALTH,FP7-HEALTH-2013-INNOVATION-1,ASKLEPIOS(2013), Institut Pasteur [Paris] (IP), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Benzylamines, Indoles, Endosome, G protein, [SDV]Life Sciences [q-bio], 030106 microbiology, Adamantane, Endosomes, Biology, medicine.disease_cause, Antiviral Agents, Late endosome, Cell Line, Viral Proteins, 03 medical and health sciences, Virology, Drug Resistance, Viral, Ribavirin, Rabies glycoprotein, medicine, Animals, Endosomal pathway inhibitors, Pharmacology, Rabies virus, RNA, Drug Synergism, Virus Internalization, Umifenovir, medicine.disease, Drug synergy, 3. Good health, 030104 developmental biology, RAB7A, Mutation, Rabies, medicine.drug
Abstract

International audience; ABMA and its analogue DABMA are two molecules of the adamantane family known to perturbate the endosomal pathway and to inhibit cell infection of several RNA and DNA viruses. Their activity against Rabies Virus (RABV) infection has already been demonstrated in vitro. (Wu et al., 2017, 2019). Here, we describe in more details their mechanism of action by comparison to Arbidol (umifenovir) and Ribavirin, two broad spectrum antivirals against emerging viruses such as Lassa, Ebola, influenza and Hantaan viruses. ABMA and DABMA, delivered 2 h pre-infection, inhibit RABV infection in vitro with an EC50 of 7.8 μM and 14 μM, respectively. They act at post-entry, by causing RABV accumulation within the endosomal compartment and DABMA specifically diminishes the expression of the GTPase Rab7a controlling the fusion of early endosomes to late endosomes or lysosomes. This may suggest that ABMA and DABMA act at different stages of the late endosomal pathway as supported by their different profile of synergy/antagonism with the fusion inhibitor Arbidol. This difference is further confirmed by the RABV mutants induced by successive passages under increasing selective pressure showing a particular involvement of the viral G protein in the DABMA inhibition while ABMA inhibition induces less mutations dispersed in the M, G and L viral proteins. These results suggest new therapeutic perspectives against rabies.

Published
2021

11. Zika virus enhances monocyte adhesion and transmigration favoring viral dissemination to neural cells

Author

Emma Partiot, Nilda Vanesa Ayala-Nunez, Jacky G. Goetz, Gautier Follain, Judith R.E. Roels, Anita Eckly, Béatrice Uring-Lambert, Maxime Chazal, Raphael Gaudin, Gillian Hale, Frank M. J. Jacobs, Sandrine Bourdoulous, Orestis Faklaris, Sherif R. Zaki, Aurélie Hirschler, Nolwenn Jouvenet, Margot Carocci, Florian Bakoa, Sarah Cianférani, Christine Carapito, Frédéric Doussau, Brigid C. Bollweg, François Delalande, Institut de Recherche en Infectiologie de Montpellier (IRIM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA), Institut National de la Santé et de la Recherche Médicale (INSERM), Immuno-Rhumatologie Moléculaire, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Strasbourg (UNISTRA), Fédération de Médecine Translationnelle de Strasbourg (FMTS), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centers for Disease Control and Prevention [Atlanta] (CDC), Centers for Disease Control and Prevention, Swammerdam Institute for Life Sciences, University of Amsterdam [Amsterdam] (UvA)-Center for Neurosciences, Génomique virale et vaccination, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Université Paris Descartes - Paris 5 (UPD5), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), BioCampus Montpellier (BCM), Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Biologie et pharmacologie des plaquettes sanguines: hémostase, thrombose, transfusion, Université de Strasbourg (UNISTRA)-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Les Hôpitaux Universitaires de Strasbourg (HUS), Institut des Neurosciences Cellulaires et Intégratives (INCI), Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), This work has received funding from the French State via the French National Research Agency (ANR) as part of the program Investissements d’avenir (IdEx Université de Strasbourg) to RG. This work was supported by an ATIP-AVENIR starting grant to RG. The research leading to these results has received funding from the People Program (Marie Curie Actions) of the European Union’s Seventh Framework Program (FP7/2007–2013) under REA grant agreement no. PCOFUND-GA-2013-609102, through the PRESTIGE program coordinated by Campus France, and from the French Agency for Research on AIDS and Viral Hepatitis (ANRS), both attributed to NVAN. Experiments conducted by G.F. and J.G.G. were funded by Plan Cancer (OptoMetaTrap), Ligue contre le Cancer, Idex Attractivités (University of Strasbourg), and by institutional funds from INSERM and University of Strasbourg. G.F. was supported by a doctoral fellowship from Ligue Contre le Cancer. Mass spectrometry experiments were supported by the French Proteomic Infrastructure (ProFI, ANR-10-INBS-08-03). F.B. is a recipient of a PhD grant provided by ANRT (Association Nationale de la Recherche et de la Technologie). N.J. is funded by Agence Nationale pour la Recherche (ANR-16-CE15-0025-01), Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur. CDC contributions to this paper written and edited by G.L.H. and B.C.B. in their private capacity. No official support or endorsement by the Centers for Disease Control and Prevention, Department of Health and Human Services is intended, nor should be inferred. All authors read, provided feedback, and approved the paper. The findings and conclusions in this report are those of the authors and do not necessarily represent the official position of the Centers for Disease Control and Prevention. The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-1 NL4-3 AD8 Infectious Molecular Clone (pNL(AD8)) from Dr. Eric O. Freed (cat# 11346)., ANR-16-CE15-0025,Viro-Storm,Mécanismes de production incontrôlée de cytokines au cours de l'infection virale(2016), European Project: 609102,EC:FP7:PEOPLE,FP7-PEOPLE-2013-COFUND,PRESTIGE(2014), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), BioCampus (BCM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), EFS-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), jouvenet, nolwenn, Mécanismes de production incontrôlée de cytokines au cours de l'infection virale - - Viro-Storm2016 - ANR-16-CE15-0025 - AAPG2016 - VALID, PRES Towards International Gain of Excellence - PRESTIGE - - EC:FP7:PEOPLE2014-09-01 - 2019-08-31 - 609102 - VALID, Equipe Direction scientifique, Sciences et Technologies de la Musique et du Son (STMS), Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-IRCAM-Centre National de la Recherche Scientifique (CNRS), École pratique des hautes études (EPHE), Neuro-Immunologie Virale, Virologie UMR1161 (VIRO), École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de Photonique Quantique et Moléculaire (LPQM), Centre National de la Recherche Scientifique (CNRS)-CentraleSupélec-École normale supérieure - Cachan (ENS Cachan), Biologie et pharmacologie des plaquettes sanguines: hémostase, thrombose, transfusion (http://www.u949.inserm.fr/), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS, Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de Masse BioOrganique, Département des Sciences Analytiques, Institut Pluridisciplinaire Hubert Curien, Strasbourg, France (LSMBO-DSA-IPHC), Centre National de la Recherche Scientifique (CNRS), Immunorhumathologie moléculaire, Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), and Molecular Neuroscience (SILS, FNWI)

Subjects
Central Nervous System, 0301 basic medicine, MESH: Neurons, General Physics and Astronomy, MESH: Organoids, MESH: Monocytes, Monocytes, Zika virus, 0302 clinical medicine, MESH: Transendothelial and Transepithelial Migration, Cerebellum, MESH: Animals, lcsh:Science, MESH: Embryonic Stem Cells, ComputingMilieux_MISCELLANEOUS, Zebrafish, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Neurons, Multidisciplinary, Zika Virus Infection, Cell adhesion molecule, food and beverages, Sciences du Vivant [q-bio]/Microbiologie et Parasitologie, Phenotype, 3. Good health, Cell biology, Organoids, Mechanisms of disease, medicine.anatomical_structure, MESH: Cell Survival, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Female, Endothelium, Cell Survival, Science, MESH: Zika Virus, Biology, MESH: Endothelium, Article, General Biochemistry, Genetics and Molecular Biology, Virus, MESH: Cell Adhesion, 03 medical and health sciences, Virology, medicine, Animals, Humans, MESH: Central Nervous System, Cellular microbiology, Cell adhesion, MESH: Zebrafish, Embryonic Stem Cells, MESH: Humans, Monocyte, fungi, Transendothelial and Transepithelial Migration, Zika Virus, General Chemistry, biology.organism_classification, Embryonic stem cell, MESH: Cerebellum, Disease Models, Animal, 030104 developmental biology, lcsh:Q, MESH: Disease Models, Animal, Cell Adhesion Molecules, MESH: Female, 030217 neurology & neurosurgery
Abstract

Zika virus (ZIKV) invades and persists in the central nervous system (CNS), causing severe neurological diseases. However the virus journey, from the bloodstream to tissues through a mature endothelium, remains unclear. Here, we show that ZIKV-infected monocytes represent suitable carriers for viral dissemination to the CNS using human primary monocytes, cerebral organoids derived from embryonic stem cells, organotypic mouse cerebellar slices, a xenotypic human-zebrafish model, and human fetus brain samples. We find that ZIKV-exposed monocytes exhibit higher expression of adhesion molecules, and higher abilities to attach onto the vessel wall and transmigrate across endothelia. This phenotype is associated to enhanced monocyte-mediated ZIKV dissemination to neural cells. Together, our data show that ZIKV manipulates the monocyte adhesive properties and enhances monocyte transmigration and viral dissemination to neural cells. Monocyte transmigration may represent an important mechanism required for viral tissue invasion and persistence that could be specifically targeted for therapeutic intervention., Zika virus (ZIKV) can infect the central nervous system, but it is not clear how it reaches the brain. Here, Ayala-Nunez et al. show in ex vivo and in vivo models that ZIKV can hitch a ride in monocytes in a Trojan Horse manner to cross the endothelium and disseminate the virus.

Published
2019

12. In vitro and in vivo evaluation of a single chain antibody fragment generated in planta with potent rabies neutralisation activity

Author

Phoolcharoen, Waranyoo, Banyard, Ashley, Préhaud, Christophe, Selden, David, Wu, Guanghui, Birch, Colin P.D., Szeto, Tim, Lafon, Monique, Fooks, Anthony, Ma, Julian K.-C., Institute for Infection and Immunity [Londres, UK], St George's, University of London, Faculty of Pharmaceutical Sciences [Bangkok], Chulalongkorn University [Bangkok], Wildlife Zoonoses and Vector-borne Diseases Research Group, Animal and Plant Health Agency [Addlestone, UK] (APHA), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Biomathematics and Risk Research Group, This work was supported by the Royal Society (Newton International fellowship to W. Phoolcharoen), a Wellcome Trust grant (WT093092MA), the Hotung Foundation and an ERC award (ERC-2010-AdG_20100317). The studies undertaken at APHA received partial financial support from the EU FP7 project (ASKLEPIOS 602825) and by the UK Department for Environment, Food and Rural Affairs (Defra projects SE0426 and SE0431)., European Project: 602825,EC:FP7:HEALTH,FP7-HEALTH-2013-INNOVATION-1,ASKLEPIOS(2013), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
N-acetylcholine receptor, Rabies, Single-chain antibody (ScFv), [SDV]Life Sciences [q-bio], Blotting, Western, Clinical disease, respiratory system, Antibodies, Neutralizing, Article, Cell Line, Mice, Rabies Vaccines, Blood-Brain Barrier, Rabies virus, Blood brain barrier (BBB), Immunoglobulin, Nicotiana benthamiana, Animals, Humans, Electrophoresis, Polyacrylamide Gel, Single-Chain Antibodies
Abstract

International audience; Rabies causes more than 60,000 human deaths annually in areas where the virus is endemic. Importantly, rabies is one of the few pathogens for which there is no treatment following the onset of clinical disease with the outcome of infection being death in almost 100% of cases. Whilst vaccination, and the combination of vaccine and rabies immunoglobulin treatment for post-exposure administration are available, no tools have been identified that can reduce or prevent rabies virus replication once clinical disease has initiated. The search for effective antiviral molecules to treat those that have already developed clinical disease associated with rabies virus infection is considered one of the most important goals in rabies research. The current study assesses a single chain antibody molecule (ScFv) based on a monoclonal antibody that potently neutralises rabies in vitro as a potential therapeutic candidate. The recombinant ScFv was generated in Nicotiana benthamiana by transient expression, and was chemically conjugated (ScFv/RVG) to a 29 amino acid peptide, specific for nicotinic acetylcholine receptor (nAchR) binding in the CNS. This conjugated molecule was able to bind nAchR in vitro and enter neuronal cells more efficiently than ScFv. The ability of the ScFv/RVG to neutralise virus in vivo was assessed using a staggered administration where the molecule was inoculated either four hours before, two days after or four days after infection. The ScFv/RVG conjugate was evaluated in direct comparison with HRIG and a potential antiviral molecule, Favipiravir (also known as T-705) to indicate whether there was greater bioavailability of the ScFv in the brains of treated mice. The study indicated that the approach taken with the ScFv/RVG conjugate may have utility in the design and implementation of novel tools targetting rabies virus infection in the brain.

Published
2019

13. Comparative analysis of neuroinvasion by Japanese encephalitis virulent and vaccine strains in an in cellulo model of human blood-brain barrier

Author

Nathalie Pardigon, Anaelle da Costa, Cécile Khou, Patricia Jeannin, Christophe Prehaud, Monique Lafon, Philippe V. Afonso, Marco Vignuzzi, Marco Aurelio Díaz-Salinas, Environnement et Risques infectieux - Environment and Infectious Risks (ERI), Institut Pasteur [Paris] (IP), Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Epidémiologie et Physiopathologie des Virus Oncogènes / Oncogenic Virus Epidemiology and Pathophysiology (EPVO (UMR_3569 / U-Pasteur_3)), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Populations virales et Pathogenèse - Viral Populations and Pathogenesis, This work was supported by a grant from the Seventh Framework Program (FP7) under grant number 278433-PREDEMICS. CK was funded by the French Ministry of Defense / Délégation Générale de l’Armement. MAD-S was funded by the DARPA INTERCEPT program (DARPA cooperative agreement #HR0011-17-2-0023., European Project: 278433,EC:FP7:HEALTH,FP7-HEALTH-2011-two-stage,PREDEMICS(2011), Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)

Subjects
0303 health sciences, Human blood, Viral encephalitis, viruses, Central nervous system, Virulence, Japanese encephalitis, Biology, medicine.disease, biology.organism_classification, Virology, Virus, 3. Good health, 03 medical and health sciences, Flavivirus, 0302 clinical medicine, medicine.anatomical_structure, Vaccine strain, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, medicine, 030217 neurology & neurosurgery, 030304 developmental biology
Abstract

Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in South East Asia. It has been suggested that JEV gets access to the central nervous system (CNS) as a consequence of a preceding inflammatory process which leads to the blood-brain barrier (BBB) disruption and viral neuroinvasion. However, what happens at early times of JEV contact with the BBB is poorly understood. In the present work, we evaluated the ability of both a virulent and a vaccine strain of JEV (JEV RP9 and SA14-14-2, respectively) to cross anin cellulohuman BBB model consisting of hCMEC/D3 human endothelial cells cultivated on permeable inserts above SK-N-SH human neuroblastoma cells. Using this system, we demonstrated that both JEV RP9 and SA14-14-2 are able to cross the BBB without disrupting it at early times post-addition. Furthermore, this BBB model was able to discriminate between the virulent RP9 and the vaccine SA14-14-2 strains, as demonstrated by the presence of almost 10 times more RP9 infectious particles that crossed the BBB than SA14-14 particles at a high MOI. Besides contributing to the understanding of early events in JEV neuroinvasion, thisin celluloBBB model represents a suitable and useful system to study the viral determinants of JEV neuroinvasiveness and the molecular mechanisms by which this flavivirus crosses the BBB at early times of neuroinvasion.

Published
2019
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14. Innovative in cellulo method as an alternative to in vivo neurovirulence test for the characterization and quality control of human live Yellow Fever virus vaccines: a pilot study

Author

Monique Lafon, Aure Saulnier, Anaelle da Costa, Cécile Khou, Christophe Prehaud, Nolwenn Nougarede, Nathalie Pardigon, Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Sanofi-Aventis R&D, SANOFI Recherche, Environnement et Risques infectieux - Environment and Infectious Risks (ERI), Institut Pasteur [Paris] (IP), This study and the fellowship of A. da Costa were funded by a grant 'Contrat de Soutien à la Recherche' provided by Sanofi Pasteur to Institut Pasteur, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]

Subjects
Quality Control, 0301 basic medicine, Vaccine safety, Pilot Projects, Bioengineering, Biology, Applied Microbiology and Biotechnology, Virus, 03 medical and health sciences, 0302 clinical medicine, In vivo, 3Rs (Replacement, medicine, Humans, Animal testing, In cellulo model, Cells, Cultured, Non-human primates, Reduction, Pharmacology, Attenuated vaccine, General Immunology and Microbiology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Viral Vaccine, Yellow Fever Vaccine, Yellow fever, Models, Immunological, General Medicine, medicine.disease, Virology, 3. Good health, Yellow Fever virus, 030104 developmental biology, Refinement), Blood-Brain Barrier, BBB-Minibrain, Vaccine, 030217 neurology & neurosurgery, Biotechnology
Abstract

International audience; Live attenuated vaccines have proved to be mostly valuable in the prevention of infectious diseases in humans, especially in developing countries. The safety and potency of vaccine, and the consistency of vaccine batch-to-batch manufacturing, must be proven before being administrated to humans. For now, the tests used to control vaccine safety largely involve animal testing. For live viral vaccines, regulations require suppliers to demonstrate the absence of neurovirulence in animals, principally in non-human primates and mice. In a search to reduce the use of animals and embracing the 3Rs principles (Replacement, Reduction, Refinement in the use of laboratory animals), we developed a new Blood-Brain Barrier Minibrain (BBB-Minibrain) in cellulo device to evaluate the neuroinvasiveness/neurovirulence of live Yellow Fever virus (YFV) vaccines. A pilot study was performed using the features of two distinct YFV strains, with the ultimate goal of proposing a companion test to characterize YFV neurovirulence. Here, we demonstrate that the BBB-Minibrain model is a promising alternative to consider for future replacement of YFV vaccine in vivo neurovirulence testing (see graphical abstract).

Published
2018

15. A cell impedance-based real-time in vitro assay to assess the toxicity of amphotericin B formulations

Author

Gillian Barratt, Stéphane Bretagne, Félix Sauvage, Aude Sturny-Leclère, Sandrine Vitry, Alexandre Alanio, Jean Menotti, Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Diderot - Paris 7 (UPD7), Université Sorbonne Paris Cité (USPC), Mycologie moléculaire, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Hôpital de la Croix-Rousse [CHU - HCL], Hospices Civils de Lyon (HCL), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Galien Paris-Sud (IGPS), Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Institut de Chimie du CNRS (INC), Université Paris-Sud - Paris 11 (UP11)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Programmed cell death, Antifungal Agents, Cell Survival, 030106 microbiology, Cell, Liposomal amphotericin B, Pharmacology, Biology, Toxicology, Proinflammatory cytokine, 03 medical and health sciences, Amphotericin B, Amphotericin B deoxycholate, Cell Adhesion, Electric Impedance, medicine, Humans, Cell adhesion, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Cell Proliferation, Aerosols, Dosage Forms, A549 cell, Proinflammatory cytokines, Epithelial Cells, Drug Combinations, medicine.anatomical_structure, Gene Expression Regulation, A549 Cells, Toxicity, Cytokines, Gene expression, In vitro assay, Cell impedance, Real-time, Deoxycholic Acid, medicine.drug
Abstract

International audience; Aerosolized liposomal amphotericin B (L-AmB) has been investigated as prophylaxis against invasive aspergil-losis. However, the clinical results are controversial and some trials suggest that toxicity could be a limitation for wider use. Our aim was to assess the dynamics of cell toxicity induced in a human alveolar epithelial cell line (A549) after exposure to LAmB (50 to 400 μg/ml) or amphotericin B deoxycholate (D-AmB; 50 to 200 μg/ml) by monitoring real-time A549 cell viability using an impedance-based technology. Results were expressed as cell index values integrating cell adhesion, proliferation, and survival. In parallel, the gene expression of proin-flammatory cytokines was quantified at 6 and 24 h after drug addition by real-time RT-PCR on cell lysates. No sustained reduction of cell indexes was observed with LAmB or empty liposomes, even at 400 μg/ml. Only the highest concentration tested of LAmB (400 μg/ml) yielded transient significant 6-fold and 4-fold induction of TNF-α and IL-8 mRNAs, respectively. In contrast, D-AmB induced a decrease in cell indexes and only the 50 μg/ ml concentration of D-AmB was followed by cell recovery, higher concentrations leading to cell death. Significant 4-fold, 7-fold and 3-fold inductions of TNF-α, IL-8 and IL-33 mRNAs were also observed at 6 h with 50 μg/ml of D-AmB. In conclusion, continuous cell impedance measurement showed no toxicity on overall cellular behavior although a slight proinflammatory cytokine expression is possible after LAmB challenge. Real-time kinetics of cell impedance is an interesting tool for initial screening of cell toxicity.

Published
2017

16. Investigation of rabies virus glycoprotein carboxyl terminus as an in vitro predictive tool of neurovirulence. A 3R approach

Author

Monique Lafon, Zakir Khan, Claude T. Sabeta, Christophe Prehaud, Wonhyo Seo, Onderstepoort Veterinary Institute (ARC - OVI), Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), University of Pretoria [South Africa], and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, MESH: Amino Acid Sequence, medicine.disease_cause, 3R, MESH: Gene Expression Regulation, Viral, MESH: Animals, MESH: Peptide Fragments, chemistry.chemical_classification, Live viral vaccine, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Viral Vaccine, Neurite outgrowth, MESH: Rabies Vaccines, 3. Good health, MESH: Rabies virus, Infectious Diseases, Gene Expression Regulation, Viral, Neurite, MESH: Rats, Recombinant Fusion Proteins, Immunology, PDZ domain, MESH: Glycoproteins, Virulence, Biology, Animal Testing Alternatives, Microbiology, Cell Line, MESH: Animal Testing Alternatives, 03 medical and health sciences, Viral Proteins, medicine, Neurites, MESH: Recombinant Fusion Proteins, Animals, Humans, PDZ, Amino Acid Sequence, Glycoproteins, MESH: Humans, 030102 biochemistry & molecular biology, Rabies virus, medicine.disease, Virology, MESH: Viral Proteins, MESH: Neurites, In vitro, Peptide Fragments, Post-licensing safety test, Rats, MESH: Cell Line, 030104 developmental biology, chemistry, Rabies Vaccines, Rabies, Glycoprotein
Abstract

International audience; In the field of live viral vaccines production, there is an unmet need for in vitro tests complying a 3R approach (Refine, Replace and Reduce the use of animal experimentation) to replace the post-licensing safety tests currently assayed in animals. Here, we performed a pilot study evaluating whether virulence of rabies virus, RABV, can be forecast by an in vitro test of neurite outgrowth. The rationale to use neurite outgrowth as a read-out for this test is based on the salient property of the cytoplasmic domain of the G-protein (Cyto-G) of virulent RABV strains - not of attenuated RABV strains - to stimulate neurite outgrowth in vitro. We observed that neurite elongation triggered by the Cyto-Gs encoded by different RABV field isolates correlate with the distinct virulence scores obtained in a mouse model of experimental rabies. Our results cast the idea that it could be feasible to predict RABV virulence by testing the in vitro property of a RABV strain to promote neurite outgrowth without the use of animal experimentation.

Published
2017

17. Comparison of G protein sequences of South African street rabies viruses showing distinct progression of the disease in a mouse model of experimental rabies

Author

Claude T. Sabeta, Alexandre Servat, Jenkins Akinbowale, Florence Cliquet, Monique Lafon, Christophe Prehaud, Wonhyo Seo, Onderstepoort Veterinary Institute (ARC - OVI), Laboratoire de la faune sauvage de Nancy, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), University of Pretoria [South Africa], Neuro-Immunologie Virale - Viral Neuro-immunology, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire de la rage et de la faune sauvage de Nancy (LRFSN), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Canid rabies, Herpestidae, Rabies, Mongoose rabies, Encephalomyelitis, Immunology, Disease, medicine.disease_cause, Neurovirulent, Microbiology, 03 medical and health sciences, Mice, South Africa, Dogs, biology.animal, Zoonoses, medicine, Animals, Humans, Dog Diseases, Gene, Lyssavirus, Phylogeny, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, 030102 biochemistry & molecular biology, biology, Virulence, Host (biology), Rabies virus, biology.organism_classification, medicine.disease, Virology, Mongoose, 3. Good health, 030104 developmental biology, Infectious Diseases, RNA, Viral
Abstract

International audience; Rabies is a fatal zoonotic disease and infections generally lead to a fatal encephalomyelitis in both humans and animals. In South Africa, domestic (dogs) and the wildlife (yellow mongoose) host species maintain the canid and mongoose rabies variants respectively. In this study, pathogenicity differences of South African canid and mongoose rabies viruses were investigated in a murine model, by assessing the progression of clinical signs and survivorship. Comparison of glycoprotein gene sequences revealed amino acid differences that may underpin the observed pathogenicity differences. Cumulatively, our results suggest that the canid rabies virus may be more neurovirulent in mice than the mongoose rabies variant.

Published
2017

18. Inhibition of polyamine biosynthesis is a broad-spectrum strategy against RNA viruses

Author

Matthew L. Albert, Marie-Pascale Frenkiel, Anavaj Sakuntabhai, Carole Tamietti, Anna Yakovleva, Marco Vignuzzi, Francis Delpeyroux, Bryan C. Mounce, Sandrine Vitry, Scott W. Werneke, Etienne Simon-Loriere, Cécile Khou, Everett Clinton Smith, Mark R. Denison, Nathalie Pardigon, Marie Flamand, Peter Jan Hooikaas, Enzo Z. Poirier, Teresa Cesaro, Gonzalo Moratorio, Matthieu Prot, Monique Lafon, Romain Volle, Giovanna Barba-Spaeth, Populations virales et Pathogenèse - Viral Populations and Pathogenesis, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Immunobiologie des Cellules Dendritiques, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Vanderbilt University Medical Center [Nashville], Vanderbilt University [Nashville], Cellule Pasteur, Université Paris Diderot - Paris 7 (UPD7)-PRES Sorbonne Paris Cité, Génétique fonctionnelle des Maladies infectieuses - Functional Genetics of Infectious Diseases, Virologie Structurale - Structural Virology, Neuro-Immunologie Virale - Viral Neuro-immunology, Biologie des virus entériques (BVE), Environnement et Risques infectieux - Environment and Infectious Risks (ERI), Institut Pasteur [Paris] (IP), This work was funded by the Laboratoire d’Excellence via Integrative Biology of Emerging Infectious Diseases (grant no. ANR-10-LABX-62-IBEID to B.C.M. and M.V.), Equipe FRM DEQ20150331759 from the French Fondation pour la Recherche Médicale (to M.V.), Institut Pasteur Defeat Dengue Program (to B.C.M. and M.V.), Erasmus Unipharma-Graduates Program (to T.C.), Amgen Scholars Program at Institut Pasteur (to A.Y.), Het Weerstandfonds (to P.J.H), NIH (grant no.R01AI108197 to M.R.D.), NIH (grant no. U19 AI109680 to M.R.D.), InstitutPasteur Programme Transversal de Recherche (grant no. ZPTR484 to F.D.), Fondation Total (grant no. S-CM15010-05B to F.D.), Roux Howard Cantarini postdoctoral fellowship (to R.V.), Seventh Framework Programme (grant no. 278433-PREDEMICS to N.P.), and Ministry of Defense, Direction Générale de l’Armement (to C.K.)., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), European Project: 278433,EC:FP7:HEALTH,FP7-HEALTH-2011-two-stage,PREDEMICS(2011), Vougny, Marie-Christine, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, Preparedness, Prediction and Prevention of Emerging Zoonotic Viruses with Pandemic Potential using Multidisciplinary Approaches - PREDEMICS - - EC:FP7:HEALTH2011-11-01 - 2016-10-31 - 278433 - VALID, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and Institut Pasteur [Paris]

Subjects
0301 basic medicine, MESH: Hemorrhagic Fever, Ebola/virology, viruses, Spermine, medicine.disease_cause, Virus Replication, Disease Outbreaks, chemistry.chemical_compound, Mice, Polyamines, Diethylnorspermine, MESH: Animals, MESH: Disease Outbreaks, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, education.field_of_study, Zika Virus Infection, MESH: Chikungunya virus/metabolism, MESH: Antiviral Agents/pharmacology, MESH: Spermine/pharmacology, Ebolavirus, MESH: Virus Replication/drug effects, 3. Good health, MESH: Chikungunya Fever/virology, MESH: Polyamines/metabolism, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, MESH: Ebolavirus/drug effects, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Zika Virus Infection/virology, Chikungunya virus, Eflornithine, [SDV.IMM] Life Sciences [q-bio]/Immunology, Immunology, Population, MESH: Hemorrhagic Fever, Ebola/drug therapy, MESH: Chikungunya virus/drug effects, Biology, Microbiology, Antiviral Agents, Virus, Cell Line, 03 medical and health sciences, MESH: Acetyltransferases/metabolism, Acetyltransferases, MESH: Mice, Inbred C57BL, Virology, Vaccines and Antiviral Agents, medicine, Animals, Humans, RNA Viruses, MESH: Zika Virus/drug effects, education, MESH: Mice, MESH: Spermine/analogs & derivatives, Ebola virus, MESH: Humans, RNA, Zika Virus, Hemorrhagic Fever, Ebola, MESH: Zika Virus Infection/drug therapy, MESH: Cell Line, Mice, Inbred C57BL, MESH: Eflornithine/pharmacology, 030104 developmental biology, MESH: RNA Viruses/drug effects, chemistry, Viral replication, Insect Science, Chikungunya Fever, MESH: Chikungunya Fever/drug therapy, Polyamine, MESH: Ebolavirus/metabolism
Abstract

RNA viruses present an extraordinary threat to human health, given their sudden and unpredictable appearance, the potential for rapid spread among the human population, and their ability to evolve resistance to antiviral therapies. The recent emergence of chikungunya virus, Zika virus, and Ebola virus highlights the struggles to contain outbreaks. A significant hurdle is the availability of antivirals to treat the infected or protect at-risk populations. While several compounds show promise in vitro and in vivo , these outbreaks underscore the need to accelerate drug discovery. The replication of several viruses has been described to rely on host polyamines, small and abundant positively charged molecules found in the cell. Here, we describe the antiviral effects of two molecules that alter polyamine levels: difluoromethylornithine (DFMO; also called eflornithine), which is a suicide inhibitor of ornithine decarboxylase 1 (ODC1), and diethylnorspermine (DENSpm), an activator of spermidine/spermine N 1 -acetyltransferase (SAT1). We show that reducing polyamine levels has a negative effect on diverse RNA viruses, including several viruses involved in recent outbreaks, in vitro and in vivo . These findings highlight the importance of the polyamine biosynthetic pathway to viral replication, as well as its potential as a target in the development of further antivirals or currently available molecules, such as DFMO. IMPORTANCE RNA viruses present a significant hazard to human health, and combatting these viruses requires the exploration of new avenues for targeting viral replication. Polyamines, small positively charged molecules within the cell, have been demonstrated to facilitate infection for a few different viruses. Our study demonstrates that diverse RNA viruses rely on the polyamine pathway for replication and highlights polyamine biosynthesis as a promising drug target.

Published
2016

19. Molecular Basis of the Interaction of the Human Protein Tyrosine Phosphatase Non-receptor Type 4 (PTPN4) with the Mitogen-activated Protein Kinase p38γ

Author

Nicolas Wolff, Edoo Bibi-Zainab, Florence Cordier, Muriel Delepierre, Célia Caillet-Saguy, Marie-Christine Vaney, Ahmed Haouz, Pierre Maisonneuve, Bertrand Raynal, Monique Lafon, Kristi Sawyer, Résonance Magnétique Nucléaire des Biomolécules, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Cellule Pasteur UPMC, Institut Pasteur [Paris]-Sorbonne Université (SU), Virologie Structurale - Structural Virology, Biophysique des macromolécules et leurs interactions, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Cristallographie (Plateforme) - Crystallography (Platform), Neuro-Immunologie Virale - Viral Neuro-immunology, This work was supported by TGIR-RMN-THC Fr3050 CNRS., Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)-Sorbonne Université (SU)

Subjects
0301 basic medicine, MESH: Cell Death, MESH: Signal Transduction, Cell signaling, AMP-activated kinase (AMPK), p38γ, MESH: Cell Line, Tumor, nuclear magnetic resonance (NMR), Phosphatase, PDZ domain, Protein tyrosine phosphatase, Biology, MESH: Multienzyme Complexes, Biochemistry, phosphatase, protein-protein interaction, 03 medical and health sciences, Mitogen-Activated Protein Kinase 12, Multienzyme Complexes, Cell Line, Tumor, MESH: Mitogen-Activated Protein Kinase 12, Humans, Protein kinase A, Molecular Biology, x-ray crystallography, MESH: Humans, MESH: Protein Tyrosine Phosphatase, Non-Receptor Type 4, 030102 biochemistry & molecular biology, Cell Death, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Kinase, Protein Tyrosine Phosphatase, Non-Receptor Type 4, Cell Biology, 3. Good health, Cell biology, 030104 developmental biology, Mitogen-activated protein kinase, PTPN4, biology.protein, Intracellular, Signal Transduction
Abstract

International audience; The human protein tyrosine phosphatase non-receptor type 4 (PTPN4) prevents cell death induction in neuroblastoma and glioblastoma cell lines in a PDZ·PDZ binding motifs-dependent manner, but the cellular partners of PTPN4 involved in cell protection are unknown. Here, we described the mitogen-activated protein kinase p38γ as a cellular partner of PTPN4. The main contribution to the p38γ·PTPN4 complex formation is the tight interaction between the C terminus of p38γ and the PDZ domain of PTPN4. We solved the crystal structure of the PDZ domain of PTPN4 bound to the p38γ C terminus. We identified the molecular basis of recognition of the C-terminal sequence of p38γ that displays the highest affinity among all endogenous partners of PTPN4. We showed that the p38γ C terminus is also an efficient inducer of cell death after its intracellular delivery. In addition to recruiting the kinase, the binding of the C-terminal sequence of p38γ to PTPN4 abolishes the catalytic autoinhibition of PTPN4 and thus activates the phosphatase, which can efficiently dephosphorylate the activation loop of p38γ. We presume that the p38γ·PTPN4 interaction promotes cellular signaling, preventing cell death induction.

Published
2016

20. Corrigendum to 'Alteration of cerebral iron metabolism in Sanfilippo syndrome' [Mol. Genet. Metab. 117 2, February 2016, p. S97]

Author

Liliane Rackowski, Cathy Gomila, Pierre-Edouard Bodet, Ahmed Hodroge, Florence Gonnet, Sandrine Vitry, Zoubida Karim, Jérôme Ausseil, Rémi Delaunay, Camille Rottier, Régis Daniel, Vincent Puy, Thibaud Lefebvre, Université de Picardie Jules Verne (UPJV), Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE - UMR 8587), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Plateforme d'Infectiologie Expérimentale (PFIE - INRA UE 1277), Institut National de la Recherche Agronomique (INRA), North Hospital, Rongeurs Sauvages, Risques Sanitaires et Gestion des Populations - UR 1233 (RS2GP), Institut National de la Recherche Agronomique (INRA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), Centre Français des Porphyries, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hopital Louis Mourier - AP-HP [Colombes], Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratory of Excellence GR-Ex ' The red cell : from genesis to death ', PRES Sorbonne Paris Cité, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS), Centre hospitalier universitaire d'Amiens (CHU Amiens-Picardie), CHU Amiens-Picardie, Mécanismes physiologiques et conséquences des calcifications cardiovasculaires: rôle des remodelages cardiovasculaires et osseux, Université de Picardie Jules Verne (UPJV)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Diderot - Paris 7 (UPD7), Institut Pasteur [Paris], Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0303 health sciences, medicine.medical_specialty, Endocrinology, Diabetes and Metabolism, 030305 genetics & heredity, Metabolism, Biology, medicine.disease, Biochemistry, 03 medical and health sciences, 0302 clinical medicine, Endocrinology, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Internal medicine, Mole, Genetics, medicine, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecular Biology, 030217 neurology & neurosurgery, Sanfilippo syndrome
Published
2016

21. Modulation of HLA-G and HLA-E Expression in Human Neuronal Cells After Rabies Virus or Herpes Virus Simplex Type 1 Infections

Author

Mireille Lafage, Philippe Moreau, Christophe Prehaud, Françoise Mégret, Nathalie Rouas-Freiss, Edgardo D. Carosella, Monique Lafon, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Service de Recherche en Hémato-Immunologie (SRHI - UMR_E 05), Université Paris Diderot - Paris 7 (UPD7)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris Diderot - Paris 7 (UPD7)

Subjects
viruses, Immunology, MESH: Neurons, Herpesvirus 1, Human, Human leukocyte antigen, Biology, medicine.disease_cause, Virus, MESH: Histocompatibility Antigens Class I, Cell Line, 03 medical and health sciences, 0302 clinical medicine, HLA-E, HLA Antigens, HLA-G, MHC class I, MESH: Up-Regulation, medicine, Humans, Immunology and Allergy, MESH: HLA Antigens, 030304 developmental biology, HLA-G Antigens, Neurons, Neurotropic virus, 0303 health sciences, MESH: Humans, Cell Membrane, Histocompatibility Antigens Class I, Rabies virus, General Medicine, Virology, MESH: Cell Line, MESH: Rabies virus, Up-Regulation, 3. Good health, MESH: Herpesvirus 1, Human, Herpes simplex virus, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, biology.protein, MESH: Cell Membrane, 030215 immunology
Abstract

Human Leukocyte Antigen (HLA)-G and E are nonclassical human MHC class I molecules. They may promote tolerance leading to virus and tumor immune escape. We recently described that the herpes simplex virus type 1 (HSV-1), a neurotropic virus inducing chronic infection and neuron latency, and rabies virus (RABV), a neuronotropic virus triggering acute neuron infection, up-regulate HLA-G expression in human neurons (NT2-N). Surface expression was only detected after RABV infection. We investigated here whether RABV and HSV-1 up-regulate HLA-E expression in human neuronal precursors (Ntera-2D/1). We found that RABV, not HSV-1, up-regulates HLA-E expression, nevertheless HLA-E could not be detected on the surface of RABV-infected Ntera-2D/1. Altogether these data suggest that HLA-G and not HLA-E could contribute to the immune escape of RABV. In contrast, there was no evidence that these molecules are used by latent HSV-1 infection. Thus, neurotropic viruses that escape the host immune response totally (RABV) or partially (HSV-1) regulate HLA-G expression on human neuronal cells differentially.

Published
2007

22. Strategies to interfere with PDZ-mediated interactions in neurons: What we can learn from the rabies virus

Author

Elouan Terrien, Nicolas Babault, Nicolas Wolff, Monique Lafon, Florence Cordier, Pierre Maisonneuve, Célia Caillet-Saguy, Florent Delhommel, Plateforme technologique de RMN biologique - Biological NMR Technological Platform, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Cellule Pasteur UPMC, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Pasteur [Paris] (IP), Neuro-Immunologie Virale, This work was supported by grants from the Institut Pasteur, Agence Nationale pour la Recherche and Institut Carnot Pasteur Maladies Infectieuses. N.B., E.T., P.M. and F.D. are recipients of fellowships from the Ministère de l’Enseignement Supérieur et de la Recherche and the Fondation pour la Recherche Médicale., Prof. Henri Buc is ackowledged for helpul discussion., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Pasteur [Paris]

Subjects
Cell signaling, G protein, Viral protein, Enzymatic regulation, MESH: PDZ Domains, [SDV]Life Sciences [q-bio], PDZ domain, Molecular Sequence Data, Biophysics, Cellular homeostasis, PDZ Domains, MESH: Amino Acid Sequence, Biology, Protein Serine-Threonine Kinases, medicine.disease_cause, 03 medical and health sciences, MESH: Rabies virus/physiology, 0302 clinical medicine, MESH: Neurons/enzymology, medicine, MESH: Protein Binding, Animals, Humans, MESH: Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, Phosphorylation, Molecular Biology, Neuronal signaling, MESH: Neurons/cytology, 030304 developmental biology, MESH: Protein-Serine-Threonine Kinases/metabolism, Neurons, 0303 health sciences, MESH: Humans, MESH: Molecular Sequence Data, Kinase, Rabies virus, Protein Tyrosine Phosphatase, Non-Receptor Type 4, MESH: Neurons/metabolism, MESH: Neurons/virology, MESH: Protein Tyrosine Phosphatase, Non-Receptor Type 4/metabolism, Cell biology, MESH: Rabies virus/metabolism, 030217 neurology & neurosurgery, Protein Binding
Abstract

International audience; PDZ (PSD-95/Dlg/ZO-1) domains play a major role in neuronal homeostasis in which they act as scaffold domains regulating cellular trafficking, self-association and catalytic activity of essential proteins such as kinases and phosphatases. Because of their central role in cell signaling, cellular PDZ-containing proteins are preferential targets of viruses to hijack cellular function to their advantage. Here, we describe how the viral G protein of the rabies virus specifically targets the PDZ domain of neuronal enzymes during viral infection. By disrupting the complexes formed by cellular enzymes and their ligands, the virus triggers drastic effect on cell signaling and commitment of the cell to either survival (virulent strains) or death (vaccinal strains). We provide structural and biological evidences that the viral proteins act as competitors endowed with specificity and affinity in an essential cellular process by mimicking PDZ binding motif of cellular partners. Disruption of critical endogenous protein-protein interactions by viral protein drastically alters intracellular protein trafficking and catalytic activity of cellular proteins that control cell homeostasis. This work opens up many perspectives to mimic viral sequences and developing innovative therapies to manipulate cellular homeostasis.

Published
2014

23. Protective role of LGP2 in influenza virus pathogenesis

Author

Pierre Charneau, Mustapha Si-Tahar, Viviane Balloy, Laurence Fiette, Michel Chignard, Julien Pothlichet, Damien Chopy, Monique Lafon, Fany Blanc, Laetitia Furio, Francina Langa, Défense innée et inflammation, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Histotechnologie et Pathologie, Institut Pasteur [Paris], Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Virologie moléculaire et Vectorologie, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], This work was supported by the Institut Pasteur (grant PTR 186), the Agence Nationale de la Recherche (grants FluProParand NKTVir to F. B. and L. F.), and the Ministère de l’Enseignement Supérieur et de la Recherche (fellowship to D. C.), ANR-07-MIME-0010,FLUPROPAR,Etude de la modulation de l'infection grippale par les protéases et les antiprotéases de l'hôte(2007), ANR-08-MIEN-0021,NKTVIR,Etude des mécanismes d'activation et rôle des lymphocytes T Natural Killer lors des infections virales(2008), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
MESH: Signal Transduction, [SDV]Life Sciences [q-bio], Gene Expression, medicine.disease_cause, [SDV.MHEP.PSR]Life Sciences [q-bio]/Human health and pathology/Pulmonology and respiratory tract, DEAD-box RNA Helicases, Mice, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Leukocytes, Influenza A virus, Immunology and Allergy, MESH: Animals, MESH: Orthomyxoviridae Infections, 0303 health sciences, MESH: RNA Helicases, Pattern recognition receptor, 3. Good health, Infectious Diseases, MESH: Survival Analysis, Host-Pathogen Interactions, DEAD Box Protein 58, [SDV.IMM]Life Sciences [q-bio]/Immunology, Inflammation Mediators, Signal transduction, medicine.symptom, RNA Helicases, Signal Transduction, Genetically modified mouse, MESH: Gene Expression, MESH: Mice, Transgenic, Orthomyxoviridae, MESH: Inflammation Mediators, MESH: Influenza A virus, Mice, Transgenic, Inflammation, Biology, MESH: Leukocytes, 03 medical and health sciences, Immune system, Orthomyxoviridae Infections, In vivo, MESH: Mice, Inbred C57BL, medicine, Animals, MESH: DEAD-box RNA Helicases, MESH: Mice, 030304 developmental biology, MESH: Host-Pathogen Interactions, biology.organism_classification, Survival Analysis, Mice, Inbred C57BL, Disease Models, Animal, Immunology, MESH: Disease Models, Animal, 030215 immunology
Abstract

International audience; Influenza A virus triggers a contagious respiratory disease that can cause considerable morbidity and mortality. Using an in vitro approach, we previously demonstrated that the pattern recognition receptor retinoic acid-inducible gene I (RIG-I) plays a key role in influenza A virus-mediated immune response. However, the importance of RIG-I signaling in vivo has not been thoroughly examined, because of the lack of an appropriate mouse models. To circumvent this issue, we generated a new transgenic mouse overexpressing LGP2 (hereafter, "LGP2 TG mice"), a major regulator of the RIG-I signaling pathway. The time course of several parameters was compared in infected wild-type and LGP2 TG mice. We found that LGP2 TG mice displayed significantly reduced inflammatory mediators and a lower leukocyte infiltration into the bronchoalveolar airspace. More importantly, LGP2 TG mice had a significant survival advantage. Hence, our in vivo study reveals that LGP2 is a major downregulator of the influenza A virus-triggered detrimental inflammatory response.

Published
2014

24. Pharmacoepidemiology studies: what levels of evidence and how can they be reached?

Author

Damien Chopy, Caroline Vigneau, Claudine Sapède, Joëlle Micallef, Maryse Lapeyre-Mestre, Muriel Malbezin, François Gueyffier, Yannick Plétan, L. Lacoin, Nicholas Moore, David Evans, Gérard Pons, Dana Morlet-Vigier, Pascal Bilbault, Patrice Verpillat, Emmanuel Oger, Patrick Blin, Sophie Muller, Epidémiologie et analyses en santé publique : risques, maladies chroniques et handicaps (LEASP), 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), Laboratoire de pharmacologie médicale et clinique, CHU Toulouse [Toulouse], Laboratoire Roche [Boulogne-Billancourt], Department of Pharmacology, Xinjiang Medical University, Laboratoire de Pharmacologie Expérimentale, Université Bordeaux Segalen - Bordeaux 2-CHU Bordeaux [Bordeaux]-Département Hospitalo-Universitaire de Pharmacologie de Bordeaux, CHU de Bordeaux Pellegrin [Bordeaux]-Université Victor Segalen - Bordeaux 2-CHU de Bordeaux Pellegrin [Bordeaux]-Université Victor Segalen - Bordeaux 2, Hôpital de Strasbourg, CHU Strasbourg, Département Hospitalo-Universitaire de Pharmacologie de Bordeaux, CHU de Bordeaux Pellegrin [Bordeaux]-Université Victor Segalen - Bordeaux 2, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire pharmaceutique Lilly France [Neuilly-sur-Seine], Evaluation et modélisation des effets thérapeutiques, Département biostatistiques et modélisation pour la santé et l'environnement [LBBE], Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Laboratoire de Biométrie et Biologie Evolutive - UMR 5558 (LBBE), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique (Inria)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS), Haute Autorité de Santé [Saint-Denis La Plaine] (HAS), Janssen-Cilag [Issy-les-Moulineaux], Euromov (EuroMov), Université de Montpellier (UM), IMS Health [Puteaux], Laboratoire GlaxoSmithKline [ Marly-le-Roi], CIC-IT Rennes, Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Pharmacologie [Rennes], CHU Pontchaillou [Rennes], Epilepsies de l'Enfant et Plasticité Cérébrale (U1129), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Neurologie et thérapeutique expérimentale, Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR70-Université Pierre et Marie Curie - Paris 6 (UPMC), Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service Pharmacologie Clinique [CHU Toulouse], Pôle Santé publique et médecine publique [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR70-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Épidémiologie et analyses en santé publique : risques, maladies chroniques et handicaps, Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Biométrie et Biologie Evolutive ( LBBE ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de Recherche en Informatique et en Automatique ( Inria ) -Centre National de la Recherche Scientifique ( CNRS ), Haute Autorité de Santé [Saint-Denis La Plaine] ( HAS ), Euromov ( EuroMov ), Centre Hospitalier Régional Universitaire [Montpellier] ( CHRU Montpellier ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Montpellier ( UM ), Hôpital Pontchaillou-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Service de Pharmacologie, Epilepsies de l'Enfant et Plasticité Cérébrale ( U1129 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Université Paris Descartes - Paris 5 ( UPD5 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -IFR70-Université Pierre et Marie Curie - Paris 6 ( UPMC ), Institut de Génétique et Développement de Rennes ( IGDR ), Université de Rennes 1 ( UR1 ), and Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -Centre National de la Recherche Scientifique ( CNRS ) -Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )

Subjects
Drug-Related Side Effects and Adverse Reactions, [SDV]Life Sciences [q-bio], Population, Scientific modelling, 030226 pharmacology & pharmacy, Toxicology, 03 medical and health sciences, 0302 clinical medicine, Medicine, Humans, Pharmacology (medical), 030212 general & internal medicine, Causation, education, Research question, education.field_of_study, Actuarial science, [ SDV ] Life Sciences [q-bio], business.industry, Pharmacoepidemiology, Evidence-based medicine, 3. Good health, Identification (information), Treatment Outcome, Epidemiologic Research Design, Evidence-Based Practice, Patient Safety, Descriptive research, business
Abstract

International audience; In pharmacoepidemiology studies, the nature of the research question will dictate the choice of methodological approach and the conditions for optimizing the level of evidence. Thus, to document the treated population and the modes of use of a new drug in real-life prescribing conditions, a descriptive approach through cross-sectional or longitudinal studies conducted on databases, or else ad-hoc studies, will be preferred. On the other hand, evaluation of the real-life "effectiveness" of a new drug will be based on cohort, case-control or scientific modeling, depending on the drug and the disease of interest. For questions involving drug risks and safety, it is the adverse effects profile that will guide the choice of study design, both for identification of the effect (signal) and assessment of causation. In all cases, in the post-marketing authorization (MA) setting, the evidence acquired in pre-MA studies serves as the basis for generating hypotheses. Whatever the research question and the method chosen to address it, the potential biases and their impact on the results need to be identified. In certain cases, a combination of several complementary approaches may prove preferable to a single study.

Published
2013

25. Ordered Phosphorylation Events in Two Independent Cascades of the PTEN C-tail Revealed by NMR

Author

Elouan Terrien, Muriel Delepierre, Alain Chaffotte, Nicolas Wolff, Christophe Prehaud, François-Xavier Theillet, Henri Buc, Monique Lafon, Florence Cordier, Résonance Magnétique Nucléaire des Biomolécules, Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Cellule Pasteur UPMC, Institut Pasteur [Paris]-Sorbonne Université (SU), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], This work was supported by the CNRS and grants from the Institut Pasteur, Agence Nationale pour la Recherche (PATHO-PDZ), and Institut Carnot Pasteur Maladies Infectieuses (NeuroVita). E.T. is a recipient of fellowships from the Ministère de l′Enseignement Supérieur et de la Recherche and from the Fondation pour la Recherche Médicale., We thank N. Kellershohn for valuable discussions, J. D’Alayer for help with mass spectrometry experiments and S. Baeriswyl for careful proofreading of the manuscript., ANR-06-MIME-0030,PATHO-PDZ,Role des proteines virales porteuses de site de fixation de domaine PDZ dans la pathogenicité virale. Analyse structure/fonction des complexes PDZ/PDZ-BS dans l'apoptose neuronale viro-induite.(2006), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Sorbonne Université (SU), and Institut Pasteur [Paris] (IP)

Subjects
MESH: Glycogen Synthase Kinase 3 beta, Resonance structures, [SDV]Life Sciences [q-bio], Peptides and proteins, Biochemistry, Catalysis, law.invention, 03 medical and health sciences, Glycogen Synthase Kinase 3, 0302 clinical medicine, Colloid and Surface Chemistry, GSK-3, law, MESH: Nuclear Magnetic Resonance, Biomolecular, MESH: PTEN Phosphohydrolase / chemistry, PTEN, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Phosphorylation, MESH: Glycogen Synthase Kinase 3 / metabolism, GSK3B, Nuclear Magnetic Resonance, Biomolecular, MESH: PTEN Phosphohydrolase / metabolism, 030304 developmental biology, 0303 health sciences, Glycogen Synthase Kinase 3 beta, MESH: Humans, biology, Cluster chemistry, MESH: Phosphorylation, Chemistry, Cell growth, Kinase, PTEN Phosphohydrolase, General Chemistry, 16. Peace & justice, Cell biology, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, 030220 oncology & carcinogenesis, biology.protein, Suppressor, Cell and molecular biology, Post-translational modification, Intracellular
Abstract

International audience; PTEN phosphatase is a tumor suppressor controlling notably cell growth, proliferation and survival. The multisite phosphorylation of the PTEN C-terminal tail regulates PTEN activity and intracellular trafficking. The dynamical nature of such regulatory events represents a crucial dimension for timing cellular decisions. Here we show that NMR spectroscopy allows reporting on the order and kinetics of clustered multisite phosphorylation events. We first unambiguously identify in vitro seven bona fide sites modified by CK2 and GSK3β kinases and two new sites on the PTEN C-terminal tail. Then, monitoring the formation of transient intermediate phosphorylated states, we determine the sequence of these reactions and calculate their apparent rate constants. Finally, we assess the dynamic formation of these phosphorylation events induced by endogenous kinases directly in extracts of human neuroblastoma cells. Taken together, our data indicate that two cascades of events controlled by CK2 and GSK3β occur independently on two clusters of sites (S380-S385 and S361-S370) and that in each cluster the reactions follow an ordered model with a distributive kinetic mechanism. Besides emphasizing the ability of NMR to quantitatively and dynamically follow post-translational modifications, these results bring a temporal dimension on the establishment of PTEN phosphorylation cascades.

Published
2012

26. Interference with the PTEN-MAST2 Interaction by a Viral Protein Leads to Cellular Relocalization of PTEN

Author

Muriel Delepierre, Nicolas Wolff, Zakir Khan, Monique Lafon, Henri Buc, Mireille Lafage, Elouan Terrien, Florence Cordier, Alain Chaffotte, Catherine Simenel, Christophe Prehaud, Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Cellule Pasteur UPMC, Institut Pasteur [Paris] (IP)-Sorbonne Université (SU), Neuro-Immunologie Virale, Institut Pasteur [Paris] (IP), This work was supported by grants from the Institut Pasteur, Agence Nationale pour la Recherche, and Institut Carnot Pasteur Maladies Infectieuses. E.T. is a recipient of fellowships from the Ministère de l’Enseignement Supérieur et de la Recherche and the Fondation pour la Recherche Médicale., We thank P. Roux (Imagopole, Institut Pasteur), P. England, B. Raynal, S. Hoos, E. Frachon, and V. Bondet (Proteopole, Institut Pasteur) for their technical expertise., Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Institut Pasteur [Paris]-Sorbonne Université (SU), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]

Subjects
Models, Molecular, MESH: Isotope Labeling, [SDV]Life Sciences [q-bio], MESH: Neurons, PDZ Domains, medicine.disease_cause, Biochemistry, 0302 clinical medicine, MESH: Nuclear Magnetic Resonance, Biomolecular, MESH: PDZ Domains, Tensin, Neurons, 0303 health sciences, biology, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Immunohistochemistry, MESH: Rabies virus, MESH: Cell Survival, Isotope Labeling, Microtubule-Associated Proteins, MESH: Models, Molecular, MESH: Spectrometry, Fluorescence, MESH: Cell Line, Tumor, Microtubule-associated protein, G protein, Viral protein, Cell Survival, Phosphatase, PDZ domain, Blotting, Western, MESH: Glycoproteins, MESH: Binding, Competitive, Calorimetry, Protein Serine-Threonine Kinases, Binding, Competitive, MESH: PTEN Phosphohydrolase, MESH: Protein-Serine-Threonine Kinases, 03 medical and health sciences, Viral Proteins, Cell Line, Tumor, medicine, PTEN, Gene silencing, Humans, MESH: Blotting, Western, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Calorimetry, Molecular Biology, Nuclear Magnetic Resonance, Biomolecular, 030304 developmental biology, Glycoproteins, MESH: Humans, PTEN Phosphohydrolase, MESH: Immunohistochemistry, Cell Biology, Molecular biology, MESH: Viral Proteins, MESH: Microtubule-Associated Proteins, Spectrometry, Fluorescence, Rabies virus, biology.protein, 030217 neurology & neurosurgery
Abstract

International audience; PTEN (phosphatase and tensin homolog deleted on chromosome 10) and MAST2 (microtubule-associated serine and threonine kinase 2) interact with each other through the PDZ domain of MAST2 (MAST2-PDZ) and the carboxyl-terminal (C-terminal) PDZ domain-binding site (PDZ-BS) of PTEN. These two proteins function as negative regulators of cell survival pathways, and silencing of either one promotes neuronal survival. In human neuroblastoma cells infected with rabies virus (RABV), the C-terminal PDZ domain of the viral glycoprotein (G protein) can target MAST2-PDZ, and RABV infection triggers neuronal survival in a PDZ-BS-dependent fashion. These findings suggest that the PTEN-MAST2 complex inhibits neuronal survival and that viral G protein disrupts this complex through competition with PTEN for binding to MAST2-PDZ. We showed that the C-terminal sequences of PTEN and the viral G protein bound to MAST2-PDZ with similar affinities. Nuclear magnetic resonance structures of these complexes exhibited similar large interaction surfaces, providing a structural basis for their binding specificities. Additionally, the viral G protein promoted the nuclear exclusion of PTEN in infected neuroblastoma cells in a PDZ-BS-dependent manner without altering total PTEN abundance. These findings suggest that formation of the PTEN-MAST2 complex is specifically affected by the viral G protein and emphasize how disruption of a critical protein-protein interaction regulates intracellular PTEN trafficking. In turn, the data show how the viral protein might be used to decipher the underlying molecular mechanisms and to clarify how the subcellular localization of PTEN regulates neuronal survival.

Published
2012

27. Peptides Targeting the PDZ Domain of PTPN4 Are Efficient Inducers of Glioblastoma Cell Death

Author

Mireille Lafage, Joseph J.B. Cockburn, Monique Lafon, Christophe Prehaud, Nicolas Wolff, Ahmed Haouz, Florence Cordier, Nicolas Babault, Félix A. Rey, Henri Buc, Muriel Delepierre, Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale, Virologie Structurale, Cristallogenèse et Diffraction des Rayons X (Plate-forme/PF6), Institut Pasteur [Paris] (IP), This work was supported by grants from the Institut Pasteur and Agence Nationale pour la Recherche. N.B. is a recipient of the Ministère de l'Education et de la Recherche fellowship., We are grateful to P. Weber (PF6, Institut Pasteur) for performing robot-driven crystallization trials and to E. Frachon and V. Bondet (PF5, Institut Pasteur) for the protein expression. We thank Soleil and SLS for providing access to synchrotron facilities and M. Bellinzoni for assistance with the data collection., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]

Subjects
Magnetic Resonance Spectroscopy, MESH: PDZ Domains, [SDV]Life Sciences [q-bio], PDZ Domains, MESH: Protein Structure, Secondary, Peptide, MESH: Flow Cytometry, Protein tyrosine phosphatase, Plasma protein binding, MESH: Amino Acid Sequence, Protein Structure, Secondary, Protein structure, Structural Biology, MESH: Glioblastoma / metabolism, MESH: Cell Death, Peptide sequence, chemistry.chemical_classification, 0303 health sciences, Cell Death, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], 030302 biochemistry & molecular biology, Protein Tyrosine Phosphatase, Non-Receptor Type 4, Flow Cytometry, MESH: Rabies virus / chemistry, 3. Good health, Cell biology, MESH: Glioblastoma / pathology, Biochemistry, MESH: Viral Proteins / pharmacology, Protein Binding, MESH: Cell Line, Tumor, PDZ domain, Phosphatase, Molecular Sequence Data, MESH: Sequence Alignment, Biology, MESH: Viral Proteins / chemical synthesis, Receptors, N-Methyl-D-Aspartate, 03 medical and health sciences, Viral Proteins, Cell Line, Tumor, Humans, Point Mutation, MESH: Protein Binding, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Amino Acid Sequence, MESH: Receptors, N-Methyl-D-Aspartate / metabolism, Molecular Biology, 030304 developmental biology, MESH: Point Mutation, MESH: Humans, MESH: Molecular Sequence Data, MESH: Magnetic Resonance Spectroscopy, C-terminus, chemistry, Rabies virus, Multiprotein Complexes, MESH: Multiprotein Complexes / metabolism, MESH: Protein Tyrosine Phosphatase, Non-Receptor Type 4 / metabolism, Glioblastoma, Peptides, Sequence Alignment, MESH: Peptides / pharmacology
Abstract

Comment in : PDZ-peptide complexes: as exciting as ever./ Trave G. Structure. 2011 Oct 12;19(10):1350-1. doi: 10.1016/j.str.2011.09.008.PMID: 22000506; International audience; PTPN4, a human tyrosine phosphatase, protects cells against apoptosis. This protection could be abrogated by targeting the PDZ domain of this phosphatase with a peptide mimicking the C-terminal sequence of the G protein of an attenuated rabies virus strain. Here, we demonstrate that glioblastoma death is triggered upon intracellular delivery of peptides, either from viral origin or from known endogenous ligands of PTPN4-PDZ, such as the C terminus sequence of the glutamate receptor subunit GluN2A. The killing efficiency of peptides closely reflects their affinities for the PTPN4-PDZ. The crystal structures of two PTPN4-PDZ/peptide complexes allow us to pinpoint the main structural determinants of binding and to synthesize a peptide of high affinity for PTPN4-PDZ enhancing markedly its cell death capacity. These results allow us to propose a potential mechanism for the efficiency of peptides and provide a target and a robust framework for the design of new pro-death compounds.

Published
2011

28. Ambivalent role of the innate immune response in rabies virus pathogenesis

Author

Françoise Mégret, Monique Lafon, Mireille Lafage, Julien Pothlichet, Mustapha Si-Tahar, Laurence Fiette, Damien Chopy, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Défense innée et inflammation, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Histopathologie humaine et Modèles animaux, Institut Pasteur [Paris], Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Institut Pasteur [Paris] (IP)

Subjects
Rabies, T cell, T-Lymphocytes, Immunology, Mice, Transgenic, medicine.disease_cause, Microbiology, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, B7-H1 Antigen, Cell Line, 03 medical and health sciences, Mice, 0302 clinical medicine, Interferon, Immunity, Virology, Cell Line, Tumor, medicine, Animals, Humans, 030304 developmental biology, Neurons, 0303 health sciences, Innate immune system, Membrane Glycoproteins, biology, Innate lymphoid cell, Rabies virus, Brain, Immunity, Innate, 3. Good health, Mice, Inbred C57BL, medicine.anatomical_structure, Insect Science, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, biology.protein, B7-1 Antigen, NIH 3T3 Cells, Pathogenesis and Immunity, Antibody, Peptides, 030217 neurology & neurosurgery, CD8, RNA Helicases, medicine.drug
Abstract

The neurotropic rabies virus (RABV) has developed several evasive strategies, including immunoevasion, to successfully infect the nervous system (NS) and trigger a fatal encephalomyelitis. Here we show that expression of LGP2, a protein known as either a positive or negative regulator of the RIG-I-mediated innate immune response, is restricted in the NS. We used a new transgenic mouse model (LGP2 TG) overexpressing LGP2 to impair the innate immune response to RABV and thus revealed the role of the RIG-I-mediated innate immune response in RABV pathogenesis. After infection, LGP2 TG mice exhibited reduced expression of inflammatory/chemoattractive molecules, beta interferon (IFN-β), and IFN-stimulated genes in their NS compared to wild-type (WT) mice, demonstrating the inhibitory function of LGP2 in the innate immune response to RABV. Surprisingly, LGP2 TG mice showed more viral clearance in the brain and lower morbidity than WT mice, indicating that the host innate immune response, paradoxically, favors RABV neuroinvasiveness and morbidity. LGP2 TG mice exhibited similar neutralizing antibodies and microglia activation to those of WT mice but showed a reduction of infiltrating CD4 + T cells and less disappearance of infiltrating CD8 + T cells. This occurred concomitantly with reduced neural expression of the IFN-inducible protein B7-H1, an immunoevasive protein involved in the elimination of infiltrated CD8 + T cells. Our study shows that the host innate immune response favors the infiltration of T cells and, at the same time, promotes CD8 + T cell elimination. Thus, to a certain extent, RABV exploits the innate immune response to develop its immunoevasive strategy.

Published
2011

29. Attenuation of rabies virulence: takeover by the cytoplasmic domain of its envelope protein

Author

Muriel Delepierre, Gene S. Tan, Françoise Mégret, Christophe Prehaud, Pauline Ménager, Matthias J. Schnell, Nicolas Wolff, Damien Chopy, Mireille Lafage, Elouan Terrien, Florence Cordier, Elodie Maitrepierre, Nicolas Babault, Monique Lafon, Patrick England, Sylviane Hoos, Henri Buc, Neuro-Immunologie Virale, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Résonance Magnétique Nucléaire des Biomolécules, Department of Microbiology and Immunology, Thomas Jefferson University-Jefferson Vaccine Centre, Biophysique des Macromolécules et de leurs Interactions, Institut Pasteur [Paris] (IP), This work was supported by grants from Institut Pasteur (IP) and Agence Nationale pour la Recherche (ANR) MIME. E.T., D.C., and N.B. are recipients of Ministère de l’Education et de la Recherche fellowships, and P.M. is a recipient of an ANR fellowship., We thank P. Roux, J. D’Alayer, V. Bondet, E. Frachon, J. Bellalou, and J.-F. Charles from IP core facilities for their help. We thank S. Loh and P. Nicotera for the gift of the SH-SY5Y cells, B. Dietzschold for fruitful discussions, E. Vinolo and B. Ruggiero for help in two-hybrid result interpretation, and F. Rey for his support., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Jefferson (Philadelphia University + Thomas Jefferson University), and Institut Pasteur [Paris]

Subjects
Cytoplasm, [SDV]Life Sciences [q-bio], MESH: Neurons, PDZ Domains, Apoptosis, Protein tyrosine phosphatase, MESH: Virulence, medicine.disease_cause, Biochemistry, Mice, 0302 clinical medicine, Viral Envelope Proteins, MESH: PDZ Domains, MESH: Animals, chemistry.chemical_classification, Neurons, 0303 health sciences, Gene knockdown, Virulence, Kinase, Protein Tyrosine Phosphatase, Non-Receptor Type 4, MESH: Amino Acid Substitution, MESH: Rabies virus, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Rabies, PDZ domain, Biology, Protein Serine-Threonine Kinases, MESH: Protein-Serine-Threonine Kinases, 03 medical and health sciences, MESH: Rabies, medicine, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Molecular Biology, MESH: Mice, 030304 developmental biology, MESH: Protein Tyrosine Phosphatase, Non-Receptor Type 4, MESH: Apoptosis, MESH: Cytoplasm, Rabies virus, Cell Biology, Molecular biology, chemistry, Amino Acid Substitution, MESH: Viral Envelope Proteins, sense organs, Glycoprotein, 030217 neurology & neurosurgery
Abstract

International audience; The capacity of a rabies virus to promote neuronal survival (a signature of virulence) or death (a marker of attenuation) depends on the cellular partners recruited by the PDZ-binding site (PDZ-BS) of its envelope glycoprotein (G). Neuronal survival requires the selective association of the PDZ-BS of G with the PDZ domains of two closely related serine-threonine kinases, MAST1 and MAST2. Here, we found that a single amino acid change in the PDZ-BS triggered the apoptotic death of infected neurons and enabled G to interact with additional PDZ partners, in particular the tyrosine phosphatase PTPN4. Knockdown of PTPN4 abrogated virus-mediated apoptosis. Thus, we propose that attenuation of rabies virus requires expansion of the set of host PDZ proteins with which G interacts, which interferes with the finely tuned homeostasis required for survival of the infected neuron.

Published
2010

30. Analyses of TET2 mutations in post-myeloproliferative neoplasm acute myeloid leukemias

Author

Eric Lippert, Isabelle Radford-Weiss, Florence Nguyen-Khac, Nathalie Nadal, M. Lafage, Dominique Penther, Francine Mugneret, Joris Andrieux, Lucile Couronné, Olivier Kosmider, Nathalie Gachard, Nicole Dastugue, Olivier Bernard, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), 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), and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cancer Research, medicine.medical_specialty, MESH: Mutation, Biology, Core Binding Factor beta Subunit, MESH: Janus Kinase 2, Dioxygenases, 03 medical and health sciences, 0302 clinical medicine, Proto-Oncogene Proteins, Internal medicine, medicine, Humans, Acute myeloid leukemias, Myeloproliferative neoplasm, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, Myeloproliferative Disorders, Hematology, MESH: Humans, Cancer, MESH: Core Binding Factor beta Subunit, Janus Kinase 2, medicine.disease, 3. Good health, DNA-Binding Proteins, MESH: Proto-Oncogene Proteins, Leukemia, Myeloid, Acute, Oncology, 030220 oncology & carcinogenesis, Mutation, Mutation (genetic algorithm), Cancer research, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Leukemia, Myeloid, Acute, MESH: DNA-Binding Proteins, MESH: Myeloproliferative Disorders
Abstract

International audience

Published
2010

31. Latent viral infections of the nervous system: Role of the host immune response

Author

Monique Lafon, Neuro-Immunologie Virale, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)

Subjects
Nervous system, viruses, Neuroimmunology, Neurones, Adaptive Immunity, CD8-Positive T-Lymphocytes, medicine.disease_cause, 0302 clinical medicine, Virus latency, Neuro-immunologie, Simplexvirus, IFN-γ, Neurons, 0303 health sciences, Latence, Bacterial Infections, Acquired immune system, HSV-1, Virus Latency, Virus, 3. Good health, medicine.anatomical_structure, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Neurology, Lytic cycle, Virus Diseases, Latency, Cytokines, Chemokines, Symbiose, Biology, Article, Interferon-gamma, 03 medical and health sciences, Immune system, TLR, medicine, Humans, Symbiosis, 030304 developmental biology, Tumor Necrosis Factor-alpha, Herpes Simplex, CD8, medicine.disease, Virology, Immunity, Innate, Herpes simplex virus, Immunology, Neurology (clinical), Nervous System Diseases, 030217 neurology & neurosurgery
Abstract

Viruses that infect the nervous system may cause acute, chronic or latent infections. Despite the so-called immunoprivileged status of the nervous system, immunosurveillance plays an important role in the fate of viral infection of the brain. Herpes simplex virus 1 (HSV-1) persists in the nervous system for the life of the host with periodic stress induced reactivation that produces progeny viruses. Prevention of reactivation requires a complex interplay between virus neurons, and immune response. New evidence supports the view that CD8+T cells employing both lytic granule- and IFN-gamma-dependent effectors are essential in setting up and maintaining HSV-1 latency. HSV-1 infection of the nervous system can be seen as a parasitic invasion which leaves the individual at risk for subsequent reactivation and disease. The recent observation that herpes virus latency may confer protection against experimental bacterial infection suggests that unexpected symbiosis may exist between latent viruses and the infected nervous system of its host.

Published
2009

32. Structural tools to design peptides promoting neuronal survival or apoptosis

Author

Wolff, Nicolas, Lafon, Monique, Cordier, Florence, Terrien, Elouan, Babault, Nicolas, Prehaud, Christophe, Lafage, Mireille, Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale, and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]
Published
2009

33. Peptides that mimic the amino-terminal end of the rabies virus phosphoprotein have antiviral activity

Author

Christophe Prehaud, Noël Tordo, Stéphanie Méhouas, Corinne Jallet, Rob W.H. Ruigrok, Guillaume Castel, Eléonore Real, Yves Jacob, M. Chteoui, Grégory Caignard, Stratégies Antivirales, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), Génomique Virale et Vaccination, Neuro-Immunologie Virale, Immunologie, génétique et traitement des maladies métaboliques et du diabète ( UMR_S 561 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Paris Descartes - Paris 5 ( UPD5 ), Laboratory of Neurogenetics and Behavior, The Rockefeller University, Génétique, Papillomavirus et Cancer Humain, Institut Pasteur [Paris], Unit for Virus Host-Cell Interactions [Grenoble] ( UVHCI ), Université Joseph Fourier - Grenoble 1 ( UJF ) -European Molecular Biology Laboratory [Grenoble] ( EMBL ) -Centre National de la Recherche Scientifique ( CNRS ), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Immunologie, génétique et traitement des maladies métaboliques et du diabète (UMR_S 561), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Rockefeller University [New York], Unit of Virus Host Cell Interactions (UVHCI), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5), Unit for Virus Host-Cell Interactions [Grenoble] (UVHCI), and Université Joseph Fourier - Grenoble 1 (UJF)-European Molecular Biology Laboratory [Grenoble] (EMBL)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH : Cell Line, viruses, MESH: Neurons, MESH: Cricetinae, Virus Replication, medicine.disease_cause, [ SDV.MP ] Life Sciences [q-bio]/Microbiology and Parasitology, Cricetinae, MESH : Rabies virus, MESH: Animals, Mononegavirales, Neurons, 0303 health sciences, biology, MESH: Peptides, MESH : Peptides, 030302 biochemistry & molecular biology, MESH : Viral Structural Proteins, MESH : Cricetinae, MESH: Rabies virus, 3. Good health, MESH : Antiviral Agents, MESH : Virus Replication, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Vesicular stomatitis virus, MESH: Antiviral Agents, MESH: Cell Line, Tumor, Immunology, MESH: Viral Structural Proteins, MESH: Two-Hybrid System Techniques, MESH: Phosphoproteins, Antiviral Agents, Microbiology, Virus, MESH : Neurons, Cell Line, MESH : Immunoprecipitation, 03 medical and health sciences, Cell Line, Tumor, Two-Hybrid System Techniques, Virology, Vaccines and Antiviral Agents, medicine, Animals, Humans, Immunoprecipitation, Lyssavirus, 030304 developmental biology, Viral Structural Proteins, MESH: Humans, MESH: Immunoprecipitation, MESH : Cell Line, Tumor, MESH: Virus Replication, Rabies virus, MESH : Humans, Rhabdoviridae, Phosphoproteins, biology.organism_classification, Molecular biology, MESH: Cell Line, Nucleoprotein, Viral replication, MESH : Two-Hybrid System Techniques, Insect Science, MESH : Animals, MESH : Phosphoproteins, Peptides, Molecular Chaperones
Abstract

We wanted to develop a therapeutic approach against rabies disease by targeting the lyssavirus transcription/replication complex. Because this complex (nucleoprotein N-RNA template processed by the L polymerase and its cofactor, the phosphoprotein P) is similar to that of other negative-strand RNA viruses, we aimed to design broad-spectrum antiviral drugs that could be used as a complement to postexposure vaccination and immunotherapy. Recent progress in understanding the structure/function of the rabies virus P, N, and L proteins predicts that the amino-terminal end of P is an excellent target for destabilizing the replication complex because it interacts with both L (for positioning onto the N-RNA template) and N (for keeping N soluble, as needed for viral RNA encapsidation). Thus, peptides mimicking various lengths of the amino-terminal end of P have been evaluated, as follows: (i) for binding properties to the N-P-L partners by the two-hybrid method; (ii) for their capacity to inhibit the transcription/replication of a rabies virus minigenome encoding luciferase in BHK-21-T7 cells; and (iii) for their capacity to inhibit rabies virus infection of BHK-21-T7 cells and of two derivatives of the neuronal SK-N-SH cell line. Peptides P60 and P57 (the first 60 and first 57 NH 2 residues of P, respectively) exhibited a rapid, strong, and long-lasting inhibitory potential on luciferase expression (>95% from 24 h to 55 h). P42 was less efficient in its inhibition level (75% for 18 to 30 h) and duration (40% after 48 h). The most promising peptides were synthesized in tandem with the Tat sequence, allowing cell penetration. Their inhibitory effects were observed on BHK-21-T7 cells infected with rabies virus and Lagos bat virus but not with vesicular stomatitis virus. In neuronal cells, a significant inhibition of both nucleocapsid inclusions and rabies virus release was observed.

Published
2009

34. Toll-like receptor 3 (TLR3) plays a major role in the formation of rabies virus Negri Bodies

Author

Mireille Lafage, Françoise Mégret, Monique Lafon, Jean-Pierre Bourgeois, Pauline Ménager, Anne-Marie Le Sourd, Pascal Roux, Christophe Prehaud, Anne Danckaert, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Imagerie Dynamique (Plate-Forme) (PFID), Institut Pasteur [Paris], Département de Neuroscience - Department of Neuroscience, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
viruses, Immunology/Innate Immunity, MESH: Neurons, Kaplan-Meier Estimate, medicine.disease_cause, Virus Replication, MESH: Mice, Knockout, Inclusion bodies, Inclusion Bodies, Viral, Mice, MESH: Cell Compartmentation, 0302 clinical medicine, MESH: Animals, MESH: Kaplan-Meiers Estimate, lcsh:QH301-705.5, Cells, Cultured, Mice, Knockout, Neurons, 0303 health sciences, Toll-like receptor, Negri bodies, virus diseases, hemic and immune systems, MESH: Toll-Like Receptor 3, 3. Good health, Cell biology, MESH: Rabies virus, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Data Interpretation, Statistical, Research Article, MESH: Cells, Cultured, lcsh:Immunologic diseases. Allergy, Rabies, MESH: Nucleocapsid, Immunology, chemical and pharmacologic phenomena, Endosomes, MESH: Microscopy, Electron, Biology, Virology/Immune Evasion, Microbiology, Virus, 03 medical and health sciences, MESH: Rabies, Virology, Genetics, medicine, Animals, Humans, Nucleocapsid, Molecular Biology, MESH: Mice, 030304 developmental biology, Innate immune system, MESH: Humans, Infectious Diseases/Infectious Diseases of the Nervous System, Rabies virus, MESH: Virus Replication, Virology/Host Invasion and Cell Entry, Cell Compartmentation, Toll-Like Receptor 3, Microscopy, Electron, Viral replication, lcsh:Biology (General), MESH: Endosomes, TLR3, MESH: Inclusion Bodies, Viral, Parasitology, lcsh:RC581-607, MESH: Data Interpretation, Statistical, 030217 neurology & neurosurgery
Abstract

Human neurons express the innate immune response receptor, Toll-like receptor 3 (TLR3). TLR3 levels are increased in pathological conditions such as brain virus infection. Here, we further investigated the production, cellular localisation, and function of neuronal TLR3 during neuronotropic rabies virus (RABV) infection in human neuronal cells. Following RABV infection, TLR3 is not only present in endosomes, as observed in the absence of infection, but also in detergent-resistant perinuclear inclusion bodies. As well as TLR3, these inclusion bodies contain the viral genome and viral proteins (N and P, but not G). The size and composition of inclusion bodies and the absence of a surrounding membrane, as shown by electron microscopy, suggest they correspond to the previously described Negri Bodies (NBs). NBs are not formed in the absence of TLR3, and TLR3−/− mice—in which brain tissue was less severely infected—had a better survival rate than WT mice. These observations demonstrate that TLR3 is a major molecule involved in the spatial arrangement of RABV–induced NBs and viral replication. This study shows how viruses can exploit cellular proteins and compartmentalisation for their own benefit., Author Summary Viruses are obligate parasites. The progression of their life cycle depends on their hijacking the cellular metabolism and machinery. Human neurons produce TLR3, a protein involved in early host defence mechanisms and the modulation of neuronal survival. Rabies virus is a neurotropic virus, infecting mainly neurons. In this study, we showed that rabies virus exploits TLR3 function to store viral proteins and viral genomic material in particular areas of the cell where virus multiplication occurs. We found that, during the course of infection, large (1–3 µm) spherical inclusions were formed within the region around the nucleus. These inclusions were composed of an inner core of aggregated TLR3 surrounded by a coat of viral proteins and genomic material. These inclusions were revealed to be the previously described Negri Bodies (NBs). In absence of TLR3, NBs were no longer formed and virus multiplication rate decreased. Mice deficient in TLR3 were more resistant to rabies and had lower levels of infection in their brains. This study shows how neurotropic viruses, such as rabies virus, hijack normal functions of neuronal proteins and use cell compartmentalisation to promote viral multiplication.

Published
2009

35. 1H, 13C and 15N resonance assignments of the PDZ of microtubule-associated serine/threonine kinase 205 (MAST205) in complex with the C-terminal motif from the rabies virus glycoprotein

Author

Muriel Delepierre, Nicolas Wolff, Christophe Prehaud, Henri Buc, Catherine Simenel, Elouan Terrien, Monique Lafon, Résonance Magnétique Nucléaire des Biomolécules, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Neuro-Immunologie Virale, Institut Pasteur [Paris], This work was supported by the Agence Nationale de la Recherche (ANR-MIME Patho-PDZ). E. Terrien is recipient of a MRT fellowship., We thanks J. D’Alayer from Institut Pasteur Facility 'Analyse et Microséquençage des Protéines' for mass spectrometry and microsequencing and V. Bondet, E. Frachon and J. Bellalou from Institut Pasteur Facility 'Production de Protéines recombinantes et d’Anticorps' for protein expression and purification., ANR-06-MIME-0030,PATHO-PDZ,Role des proteines virales porteuses de site de fixation de domaine PDZ dans la pathogenicité virale. Analyse structure/fonction des complexes PDZ/PDZ-BS dans l'apoptose neuronale viro-induite.(2006), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
Magnetic Resonance Spectroscopy, Amino Acid Motifs, MESH: Amino Acid Sequence, medicine.disease_cause, Biochemistry, Viral infection, MESH: Amino Acid Motifs, MESH: Protein Structure, Tertiary, 0302 clinical medicine, Structural Biology, chemistry.chemical_classification, 0303 health sciences, Carbon Isotopes, 3. Good health, MESH: Rabies virus, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Protons, Microtubule-Associated Proteins, MESH: Nitrogen Isotopes, Protein Binding, PDZ domain, Molecular Sequence Data, MESH: Glycoproteins, Biology, Protein Serine-Threonine Kinases, MESH: Protein-Serine-Threonine Kinases, 03 medical and health sciences, Viral Proteins, Microtubule, medicine, MESH: Protein Binding, Amino Acid Sequence, 030304 developmental biology, Glycoproteins, Serine/threonine-specific protein kinase, MESH: Molecular Sequence Data, Binding Sites, Nitrogen Isotopes, MESH: Magnetic Resonance Spectroscopy, Rabies virus, MESH: Carbon Isotopes, MESH: Multiprotein Complexes, MESH: Viral Proteins, Protein Structure, Tertiary, MESH: Microtubule-Associated Proteins, chemistry, MESH: Binding Sites, Multiprotein Complexes, MESH: Protons, Glycoprotein, 030217 neurology & neurosurgery
Abstract

International audience; Most of microbes hijack the cellular machinery to their advantage by interacting with specific target of the host cell. Glycoprotein of rabies virus is a key factor controlling the homeostasis of infected neuronal cells and proteins belonging to the human microtubule associated serine threonine kinase family have been identified as potential cellular partners. As a first step towards its structural study, we have assigned the backbone and side chain nuclei resonances of the PDZ domain (PSD-95, Discs Large, ZO-1) of MAST205 in complex with the C-terminal residues of the glycoprotein of rabies virus. The BMRB accession code is 155972.

Published
2008

36. High CRMP2 expression in peripheral T lymphocytes is associated with recruitment to the brain during virus-induced neuroinflammation

Author

Pascale Giraudon, Monique Lafon, I. Sagardoy, Sylvie Cavagna, I. Chounlamountri, M. Varrin-Doyer, Carine Vuaillat, Arlette Bernard, INSERM IFR19, Institut National de la Santé et de la Recherche Médicale (INSERM), Neuro-oncologie et neuro-inflammation, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Baran, Corinne

Subjects
Antigens, Differentiation, T-Lymphocyte, CD3 Complex, Rabies, CD3, Lymphocyte, T-Lymphocytes, Immunology, Nerve Tissue Proteins, Biology, medicine.disease_cause, Virus, 03 medical and health sciences, Mice, 0302 clinical medicine, Antigens, CD, Cell Movement, medicine, Immunology and Allergy, Animals, Lectins, C-Type, Encephalitis, Viral, Distemper, Neurovirology, Neuroinflammation, 030304 developmental biology, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, 0303 health sciences, Mice, Inbred BALB C, Rabies virus, Brain, Virology, 3. Good health, Neuroimmunology, medicine.anatomical_structure, Neurology, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, biology.protein, Intercellular Signaling Peptides and Proteins, Female, Neurology (clinical), Collapsin response mediator protein family, 030217 neurology & neurosurgery
Abstract

Collapsin Response Mediator Protein (CRMP)-2 is involved in T-cell polarization and migration. To address the role of CRMP2 in neuroinflammation, we analyzed its involvement in lymphocyte recruitment to the central nervous system in mouse infected with neurotropic and non-neurotropic virus strains (RABV, CDV). A sub-population of early-activated CD69+CD3+ T lymphocytes highly expressing CRMP2 (CRMP2hi) peaked in the blood, lymph nodes and brain of mice infected with neurotropic viruses, and correlated with severity of disease. They displayed high migratory properties reduced by CRMP2 blocking antibody. These data point out the potential use of CRMP2 as a peripheral indicator of neuroinflammation.

Published
2008

37. Apoptosis inversely correlates with rabies virus neurotropism

Author

Mireille Lafage, Santos A. Susin, Nicole Israël, Guido Kroemer, Maria-Isabel Thoulouze, Monique Lafon, Victor J. Yuste, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Biologie des Interactions Cellulaires (BIC), Apoptose et Système Immunitaire (ASI), Génétique oncologique [Villejuif], Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Biologie des Rétrovirus, Institut Pasteur [Paris], Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Institut Pasteur [Paris] (IP)

Subjects
T cell, Cell, Apoptosis, medicine.disease_cause, Jurkat cells, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Jurkat Cells, MESH: Brain, History and Philosophy of Science, medicine, MESH: Jurkat Cells, Humans, MESH: Flavoproteins, Caspase, 030304 developmental biology, Neurotropic virus, 0303 health sciences, MESH: Caspases, MESH: Humans, biology, Flavoproteins, 030306 microbiology, General Neuroscience, MESH: Apoptosis, Rabies virus, Apoptosis Inducing Factor, Brain, Membrane Proteins, hemic and immune systems, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, medicine.disease, Virology, 3. Good health, MESH: Rabies virus, medicine.anatomical_structure, MESH: Apoptosis Inducing Factor, Proto-Oncogene Proteins c-bcl-2, MESH: Proto-Oncogene Proteins c-bcl-2, Caspases, biology.protein, Rabies, MESH: Membrane Proteins
Abstract

We report that non-neurotropic rabies virus (RV) strains, currently used to immunize wildlife against rabies, induces not only a caspase-dependent apoptosis in the human lymphoblastoid Jurkat T cell line (Jurkat-vect), but also a caspase-independent pathway. Cell redistribution of the apoptosis-inducing factor (AIF) was observed in Jurkat-vect infected with RV vaccine strain. Bcl-2 overproduction in Jurkat T cells (Jurkat-Bcl-2) abolished both caspase activation and AIF distribution. In contrast, strain of neurotropic RV did not induce apoptosis. The inverse correlation of the induction of apoptosis and the capacity of a virus strain to invade the brain suggests that blockage of apoptosis could be a strategy selected by neurotropic virus to favor its progression through the nervous system.

Published
2003

38. Influence of mixed chimerism on the results of allogeneic bone marrow transplantation for leukemia

Author

Pierre Levy, Didier Blaise, M. Lafage, Anne-Marie Stoppa, Patrice Mannoni, Patrice Viens, MF Bertheas, Dominique Maraninchi, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Georgia Institute of Technology [Lorraine, France], 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), Bertheas MF, Lafage M, Levy P, Blaise D, Stoppa AM, Viens P, Mannoni P, Maraninchi D., and Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)

Subjects
medicine.medical_specialty, MESH: Graft Survival, T-Lymphocytes, Immunology, Graft vs Host Disease, MESH: Graft vs Host Disease, Biochemistry, Lymphocyte Depletion, 03 medical and health sciences, Chimera (genetics), Myelogenous, 0302 clinical medicine, Leukemia, Myelogenous, Chronic, BCR-ABL Positive, MESH: Leukemia, medicine, Humans, MESH: Bone Marrow Transplantation, MESH: Chimera, 030304 developmental biology, Bone Marrow Transplantation, MESH: Lymphocyte Depletion, 0303 health sciences, Leukemia, MESH: Humans, Chimera, business.industry, Graft Survival, Cytogenetics, Myeloid leukemia, Cell Biology, Hematology, Total body irradiation, medicine.disease, 3. Good health, Haematopoiesis, medicine.anatomical_structure, MESH: T-Lymphocytes, MESH: Leukemia, Myelogenous, Chronic, BCR-ABL Positive, 030220 oncology & carcinogenesis, Acute Disease, MESH: Acute Disease, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Bone marrow, business, 030215 immunology
Abstract

Serial cytogenetic studies were performed on 60 leukemic recipients of HLA-matched bone marrow transplants (BMT) who were prepared by high doses of alkylating agents and fractionated total body irradiation (TBI). Forty-three patients were recipients of untreated BMT and 17 were recipients of T-depleted BMT. Donor or host mitoses were identified by examination of sex chromosomes in 54 patients or by evaluation of the polymorphism of other chromosomes after specific banding in six patients. Mixed lymphoid chimerism (MLC) was identified in 29 patients and full donor lymphoid chimerism (FDLC) in 29 patients. Complete donor hematopoiesis was recovered in most patients after 12 months, but two T-depleted patients had persistent host cells at 46 and 52 months after the graft. Acute graft-versus-host disease was significantly less frequent in patients with MLC, especially when more than 10% of residual lymphoid cells were detected. The probability of relapse and survival was identical in patients with MLC and FDLC, except in patients with chronic myeloid leukemia where MLC was significantly associated with an increased risk of relapse.

Published
1991

39. Isolation and mapping of a polymorphic DNA sequence (pB3.811) on chromosome 12 [D12S33]

Author

Rouault, jean-pierre, Rimokh, R., Rouault, J.P., Wahbi, K., Gadoux, M., Lafage, M., Cornillet, P., Samarut, J., Germain, D., Magaud, J.P., Communications Cellulaires et Différenciation (CCD), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Recherche Agronomique (INRA), Institut des Sciences Pharmaceutiques et Biologiques (ISPB), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon, Neuro-Immunologie Virale, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut de biologie et chimie des protéines [Lyon] (IBCP), and Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)

Subjects
Genetics, Chromosomes, Human, Pair 12, Polymorphism, Genetic, Base Sequence, [SDV]Life Sciences [q-bio], Breakpoint, Chromosome, Chromosome Mapping, Chromosomal translocation, DNA, Biology, Molecular biology, DNA sequencing, PstI, Gene mapping, biology.protein, Humans, Restriction fragment length polymorphism, Chromosome 12, Polymorphism, Restriction Fragment Length, ComputingMilieux_MISCELLANEOUS
Abstract

A 0.45 kb HindIII genomic fragment subclones into pUC18. This fragment was isolated from a genomic phage insert containing the breakpoint of a t(8;12)(q24,q22-23) chromosomal translocation. PstI identifies a simple two alleles polymorphism at either 5.5 kb or 2 kb, without constant band. The probe has been assigned to chromosome 12q22-23 by in situ hybridization studies. Co-dominant segregation of the PstI RFLP was observed in 6 families (17 informatives meiosis).

Published
1989

40. Detrimental contribution of the immuno-inhibitor B7-H1 to rabies virus encephalitis

Author

Françoise Mégret, Sven G. Meuth, Lena Alexopoulou, Myriam L Velandia Romero, Mireille Lafage, Christophe Prehaud, Heinz Wiendl, Lieping Chen, Ole J. Simon, Monique Lafon, Richard A. Flavell, Neuro-Immunologie Virale, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Department of Neurology, University of Würzburg = Universität Würzburg, Instituto de Virologia, Universidad El Bosque [Bogota], Department of Dermatology and Oncology, Johns Hopkins University School of Medicine, 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), Section of Immunobiology, Yale School of Medicine [New Haven, Connecticut] (YSM), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU), Yale University School of Medicine, and Velandia-Romero, Myriam Lucía [0000-0002-3340-7304]

Subjects
Rabies, Immunology, Biology, Virus ARN, medicine.disease_cause, Virus de la rabia, Virus, B7-H1 Antigen, 03 medical and health sciences, Interferon-gamma, Mice, 0302 clinical medicine, Immune system, Antigens, CD, Cell Line, Tumor, medicine, Immunology and Allergy, Cytotoxic T cell, Animals, Humans, Encephalitis, Viral, Virus de la encefalitis, Cells, Cultured, 030304 developmental biology, Neurotropic virus, Neurons, 0303 health sciences, Membrane Glycoproteins, Tumor Necrosis Factor-alpha, Rabies virus, Brain, Interferon-beta, medicine.disease, Virology, Mice, Mutant Strains, 3. Good health, Spinal Cord, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, B7-1 Antigen, Peptides, Encephalitis, CD8, 030215 immunology
Abstract

Rabies virus is the etiological agent of an acute encephalitis, which in absence of post exposure treatment is fatal in almost all cases. Virus lethality rests on its ability to evade the immune response. In this study, we analyzed the role of the immuno-inhibitory molecule B7-H1 in this virus strategy. We showed that in the brain and spinal cord of mice, rabies virus infection resulted in significant up-regulation of B7-H1 expression, which is specifically expressed in infected neurons. Correlatively, clinical rabies in B7-H1−/− mice is markedly less severe than in wild-type mice. B7-H1−/− mice display resistance to rabies. Virus invasion is reduced and the level of migratory CD8 T cells increases into the nervous system, while CD4/CD8 ratio remains unchanged in the periphery. In vivo, neuronal B7-H1 expression is critically depending on TLR3 signaling and IFN-β, because TLR3−/− mice—in which IFN-β production is reduced—showed only a limited increase of B7-H1 transcripts after infection. These data provide evidence that neurons can express the B7-H1 molecule after viral stress or exposure to a particular cytokine environment. They show that the B7-H1/PD-1 pathway can be exploited locally and in an organ specific manner—here the nervous system—by a neurotropic virus to promote successful host invasion.

41. Comparative analysis of neuroinvasion by Japanese encephalitis virulent and vaccine viral strains in an in vitro model of human blood-brain barrier.

Author

Khou C, Díaz-Salinas MA, da Costa A, Préhaud C, Jeannin P, Afonso PV, Vignuzzi M, Lafon M, and Pardigon N

Subjects
Blood-Brain Barrier physiology, Cell Line, Encephalitis Virus, Japanese genetics, Encephalitis Virus, Japanese pathogenicity, Encephalitis, Japanese pathology, Encephalitis, Japanese virology, Endothelial Cells cytology, Endothelial Cells metabolism, Endothelial Cells virology, Humans, RNA, Viral genetics, RNA, Viral metabolism, Virulence, Virus Replication, Blood-Brain Barrier virology, Encephalitis Virus, Japanese physiology, Models, Biological
Abstract

Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in South East Asia. It has been suggested that, as a consequence of the inflammatory process during JEV infection, there is disruption of the blood-brain barrier (BBB) tight junctions that in turn allows the virus access to the central nervous system (CNS). However, what happens at early times of JEV contact with the BBB is poorly understood. In the present work, we evaluated the ability of both a virulent and a vaccine strain of JEV (JEV RP9 and SA14-14-2, respectively) to cross an in vitro human BBB model. Using this system, we demonstrated that both JEV RP9 and SA14-14-2 are able to cross the BBB without disrupting it at early times post viral addition. Furthermore, we find that almost 10 times more RP9 infectious particles than SA14-14 cross the model BBB, indicating this BBB model discriminates between the virulent RP9 and the vaccine SA14-14-2 strains of JEV. Beyond contributing to the understanding of early events in JEV neuroinvasion, we demonstrate this in vitro BBB model can be used as a system to study the viral determinants of JEV neuroinvasiveness and the molecular mechanisms by which this flavivirus crosses the BBB during early times of neuroinvasion., Competing Interests: The authors have declared that no competing interests exist.

Published
2021
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42. Botulinum neurotoxin type B uses a distinct entry pathway mediated by CDC42 into intestinal cells versus neuronal cells.

Author

Connan C, Voillequin M, Chavez CV, Mazuet C, Leveque C, Vitry S, Vandewalle A, and Popoff MR

Subjects
Animals, Mice, Botulinum Toxins, Type A metabolism, Endocytosis, Epithelial Cells metabolism, Neurons metabolism, cdc42 GTP-Binding Protein metabolism
Abstract

Botulinum neurotoxins (BoNTs) are responsible for severe flaccid paralysis by inhibiting the release of acetylcholine at the neuromuscular junctions. BoNT type B (BoNT/B) most often induces mild forms of botulism with predominant dysautonomic symptoms. In food borne botulism and botulism by intestinal colonisation such as infant botulism, which are the most frequent naturally acquired forms of botulism, the digestive tract is the main entry route of BoNTs into the organism. We previously showed that BoNT/B translocates through mouse intestinal barrier by an endocytosis-dependent mechanism and subsequently targets neuronal cells, mainly cholinergic neurons, in the intestinal mucosa and musculosa. Here, we investigated the entry pathway of BoNT/B using fluorescent C-terminal domain of the heavy chain (HcB), which is involved in the binding to specific receptor(s) and entry process into target cells. While the combination of gangliosides GD 1a /GD 1b /GT 1b and synaptotagmin I and to a greater extent synaptotagmin II constitutes the functional HcB receptor on NG108-15 neuronal cells, HcB only uses the gangliosides GD 1a /GD 1b /GT 1b to efficiently bind to m-IC cl2 intestinal cells. HcB enters both cell types by a dynamin-dependent endocytosis, which is efficiently prevented by Dynasore, a dynamin inhibitor, and reaches a common early endosomal compartment labeled by early endosome antigen (EEA1). In contrast to neuronal cells, HcB uses a Cdc42-dependent pathway to enter intestinal cells. Then, HcB is transported to late endosomes in neuronal cells, whereas it exploits a nonacidified pathway from apical to basal lateral side of m-IC cl2 cells supporting a transcytotic route in epithelial intestinal cells., (© 2017 John Wiley & Sons Ltd.)

Published
2017
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43. Molecular Basis of the Interaction of the Human Protein Tyrosine Phosphatase Non-receptor Type 4 (PTPN4) with the Mitogen-activated Protein Kinase p38γ.

Author

Maisonneuve P, Caillet-Saguy C, Vaney MC, Bibi-Zainab E, Sawyer K, Raynal B, Haouz A, Delepierre M, Lafon M, Cordier F, and Wolff N

Subjects
Cell Death, Cell Line, Tumor, Humans, Mitogen-Activated Protein Kinase 12 genetics, Multienzyme Complexes genetics, Protein Tyrosine Phosphatase, Non-Receptor Type 4 genetics, Mitogen-Activated Protein Kinase 12 metabolism, Multienzyme Complexes metabolism, Protein Tyrosine Phosphatase, Non-Receptor Type 4 metabolism, Signal Transduction physiology
Abstract

The human protein tyrosine phosphatase non-receptor type 4 (PTPN4) prevents cell death induction in neuroblastoma and glioblastoma cell lines in a PDZ·PDZ binding motifs-dependent manner, but the cellular partners of PTPN4 involved in cell protection are unknown. Here, we described the mitogen-activated protein kinase p38γ as a cellular partner of PTPN4. The main contribution to the p38γ·PTPN4 complex formation is the tight interaction between the C terminus of p38γ and the PDZ domain of PTPN4. We solved the crystal structure of the PDZ domain of PTPN4 bound to the p38γ C terminus. We identified the molecular basis of recognition of the C-terminal sequence of p38γ that displays the highest affinity among all endogenous partners of PTPN4. We showed that the p38γ C terminus is also an efficient inducer of cell death after its intracellular delivery. In addition to recruiting the kinase, the binding of the C-terminal sequence of p38γ to PTPN4 abolishes the catalytic autoinhibition of PTPN4 and thus activates the phosphatase, which can efficiently dephosphorylate the activation loop of p38γ. We presume that the p38γ·PTPN4 interaction promotes cellular signaling, preventing cell death induction., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)

Published
2016
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44. Latent viral infections of the nervous system: role of the host immune response.

Author

Lafon M

Subjects
Adaptive Immunity, Bacterial Infections prevention & control, CD8-Positive T-Lymphocytes immunology, Chemokines immunology, Cytokines immunology, Humans, Immunity, Innate, Interferon-gamma metabolism, Simplexvirus isolation & purification, Symbiosis, Tumor Necrosis Factor-alpha immunology, Virus Latency, Herpes Simplex immunology, Nervous System Diseases immunology, Nervous System Diseases virology, Virus Diseases immunology
Abstract

Viruses that infect the nervous system may cause acute, chronic or latent infections. Despite the so-called immunoprivileged status of the nervous system, immunosurveillance plays an important role in the fate of viral infection of the brain. Herpes simplex virus 1 (HSV-1) persists in the nervous system for the life of the host with periodic stress induced reactivation that produces progeny viruses. Prevention of reactivation requires a complex interplay between virus neurons, and immune response. New evidence supports the view that CD8+T cells employing both lytic granule- and IFN-gamma-dependent effectors are essential in setting up and maintaining HSV-1 latency. HSV-1 infection of the nervous system can be seen as a parasitic invasion which leaves the individual at risk for subsequent reactivation and disease. The recent observation that herpes virus latency may confer protection against experimental bacterial infection suggests that unexpected symbiosis may exist between latent viruses and the infected nervous system of its host.

Published
2009
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45. Dynamics of immune cell recruitment during West Nile encephalitis and identification of a new CD19+B220-BST-2+ leukocyte population.

Author

Bréhin AC, Mouriès J, Frenkiel MP, Dadaglio G, Desprès P, Lafon M, and Couderc T

Subjects
Animals, Antigen-Presenting Cells immunology, Antigen-Presenting Cells metabolism, B-Lymphocytes immunology, B-Lymphocytes virology, Cell Lineage, Central Nervous System cytology, Central Nervous System immunology, Central Nervous System virology, Dendritic Cells immunology, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Fluorescent Antibody Technique, GPI-Linked Proteins, Histocompatibility Antigens Class II metabolism, Killer Cells, Natural immunology, Leukocytes immunology, Lymph Nodes cytology, Lymph Nodes immunology, Macrophages immunology, Mice, Microglia immunology, T-Lymphocytes immunology, T-Lymphocytes virology, Antigens, CD metabolism, Antigens, CD19 metabolism, Chemotaxis, Leukocyte immunology, Leukocyte Common Antigens metabolism, Leukocytes metabolism, Membrane Glycoproteins metabolism, West Nile Fever immunology
Abstract

West Nile virus (WNV) is an emerging neurotropic flavivirus. We investigated the dynamics of immune cell recruitment in peripheral tissues and in the CNS during WNV encephalitis in an immunocompetent mouse model. In the periphery, immune cell expansion can successfully limit viremia and lymphoid tissue infection. However, viral clearance in the periphery is too late to prevent viral invasion of the CNS. In the CNS, innate immune cells, including microglia/macrophages, NK cells, and plasmacytoid dendritic cells, greatly expand as the virus invades the brain, whereas B and T cells are recruited after viral invasion, and fail to control the spread of the virus. Thus, the onset of WNV encephalitis was correlated both with CNS viral infection and with a large local increase of innate immune cells. Interestingly, we identify a new immune cell type: CD19(+)B220(-) BST-2(+), which we name G8-ICs. These cells appear during peripheral infection and enter the CNS. G8-ICs express high levels of MHC class II, stain for viral Ag, and are localized in the paracortical zone of lymph nodes, strongly suggesting they are previously unidentified APCs that appear in response to viral infection.

Published
2008
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