Versailles Saint-Quentin-en-Yvelines University 4 articles published in JoVE Immunology and Infection Generation, Amplification, and Titration of Recombinant Respiratory Syncytial Viruses Camille Bouillier1, Vincent Rincheval1, Delphine Sitterlin1, Sabine Blouquit-Laye1, Aurore Desquesnes1, Jean-François Eléouët2, Elyanne Gault1,3, Marie-Anne Rameix-Welti1,3 1UMR 1173 Institut national de la santé et de la recherche médicale (INSERM), Université de Versailles St. Quentin, 2UR892 Institut national de la recherche agronomique (INRA), Unité de virologie et immunologie moléculaires, 3Assistance Publique–Hôpitaux de Paris (AP-HP), Hôpital Ambroise Paré, Laboratoire de Microbiologie We describe a method for generating and amplifying genetically modified respiratory syncytial viruses (RSVs) and an optimized plaque assay for RSVs. We illustrate this protocol by creating two recombinant viruses that respectively allow quantification of RSV replication and live analysis of RSV inclusion bodies and inclusion bodies-associated granules dynamics. Genetics Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions Charles Decraene1,2, Amanda Bortolini Silveira1, Marc Michel1, François-Clément Bidard1,3,4, Jean-Yves Pierga1,3,5, Marc-Henri Stern1,6, Charlotte Proudhon1 1Circulating Tumor Biomarkers Laboratory, SiRIC, Translational Research Department, Institut Curie, PSL Research University, 2CNRS UMR144, Institut Curie, PSL Research University, 3Department of Medical Oncology, Institut Curie, PSL Research University, 4University Versailles Saint-Quentin-en-Yvelines, 5University Paris Descartes, 6Inserm U830, Institut Curie, PSL Research University Here, we present a protocol to accurately quantify multiple genetic alterations of a target region in a single reaction using drop-off ddPCR and a unique pair of hydrolysis probes. Immunology and Infection Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos Audrey Bernut1,2, Christian Dupont1,2, Alain Sahuquet1, Jean-Louis Herrmann3, Georges Lutfalla1, Laurent Kremer1,2 1Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS, UMR 535, Université Montpellier, 2Centre d'études d'agents Pathogènes et Biotechnologies pour la Santé, CNRS, FRE 3689, Université Montpellier, 3Unité de Formation et de Recherche des Sciences de la Santé, EA3647-EPIM, Université Versailles St Quentin Optically transparent zebrafish embryos are widely used to study and visualize in real time the interactions between pathogenic microorganisms and the innate immune cells. Micro-injection of Mycobacterium abscessus, combined with fluorescence imaging, is used to scrutinize essential pathogenic features such as cord formation in zebrafish embryos. Medicine A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity Emilie Keomani1, Thérèse B. Deramaudt1, Michel Petitjean1,2, Marcel Bonay1,2, Frédéric Lofaso1,3, Stéphane Vinit1 1UFR des sciences de la santé - Simone Veil, Université de Versailles Saint-Quentin-en-Yvelines, 2Service de Physiologie - Explorations fonctionnelles, Hôpital Ambroise Paré, 3Services de Physiologie, Explorations Fonctionnelles, Réanimation Médicale et Centre d'Investigation Clinique et d'Innovation Technologique (Unité Inserm 805), Université de Versailles Saint-Quentin-en-Yvelines Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.
