NNT : 2016SACLV016 THESE DE DOCTORAT DE L’UNIVERSITE PARIS-SACLAY PREPAREE A L’UNIVERSITE VERSAILLES SAINT-QUENTIN EN YVELINES ECOLE DOCTORALE N° 577 STRUCTURE ET DYNAMIQUE DES SYSTEMES VIVANTS SCIENCES DE LA VIE ET DE LA SANTE Par Mlle Elina Aleksejeva Défenses innées antivirales du poisson zèbre: de la signalisation aux cellules spécialisées Antiviral innate defense in zebrafish: from signalling to specialized cells Thèse présentée et soutenue à Jouy-en-Josas, le 20. Janvier, 2016: Composition du Jury : M. Herrmann, Jean-Louis PUPH INSERM, Hopital Raymond-Poincaré Président M. Lutfalla, Georges DR CNRS, Universite Montpellier 2 Rapporteur M. Nisole, Sébastien CR INSERM, Universite Paris Descartes Rapporteur Mme. Vriz, Sophie PR INSERM, CNRS, College de France Examinatrice M. Boudinot, Pierre DR INRA Jouy-en-Josas Directeur de thèse M. Levraud, Jean-Pierre DR CNRS, Institut Pasteur Co-directeur de thèse Table of contents Acknowledgements .......................................................................................................................................... 1 Preamble ........................................................................................................................................................... 2 INTRODUCTION ........................................................................................................................................... 6 CHAPTER I................................................................................................................................................... 7 Type I IFN system of mammals and fish ...................................................................................................... 7 Virus sensing and conserved signalling pathways leading to type I IFN production ............................... 8 Type I IFNs and their receptors .............................................................................................................. 13 Induction pathways of ISGs ..................................................................................................................... 15 ISG repertoire .......................................................................................................................................... 17 CHAPTER II ............................................................................................................................................... 22 PLZF transcription factor – an emerging regulator of innate responses to pathogens ................................ 22 The family of BTB/POZ transcription factors ......................................................................................... 23 Promyelocytic leukemia zinc finger (PLZF) or ZBTB16 ......................................................................... 24 PLZF protein domain-function analysis .................................................................................................. 24 Intracellular shuffling of PLZF in the context of cell cycle regulation ................................................... 26 Biological functions ................................................................................................................................. 27 PLZF involvement in anti-viral and-bacterial defence ........................................................................... 29 Fish PLZF ................................................................................................................................................ 30 CHAPTER III .............................................................................................................................................. 32 TRIM antiviral immunity in mammals and fish .......................................................................................... 32 TRIM proteins – RING-B-box-CC superfamily ....................................................................................... 33 Protein domains and their functions ....................................................................................................... 33 Intracellular localization ......................................................................................................................... 35 TRIM in antiviral innate immunty ........................................................................................................... 36 TRIMs in fish and mammals: different repertoires .................................................................................. 38 The finTRIM subset: a large fish-specific group of B30.2 TRIM proteins .............................................. 39 CHAPTER IV .............................................................................................................................................. 41 In vivo studies of leukocytes in zebrafish .................................................................................................... 41 Zebrafish immune system : general anatomy .......................................................................................... 42 Zebrafish leukocytes ................................................................................................................................ 43 RESULTS ....................................................................................................................................................... 48 Aims of the thesis and research projects ..................................................................................................... 49 CHAPTER V ............................................................................................................................................... 52 Zebrafish Plzf transcription factors augment type I IFN ............................................................................. 52 Supplementary materials ......................................................................................................................... 62 CHAPTER VI ............................................................................................................................................. 63 Steady state tissue-specific antiviral immunity: a finTRIM inducing constitutive IFN signaling is expressed at surfaces exposed to pathogens ................................................................................................ 63 Abstract.................................................................................................................................................... 65 Introduction ............................................................................................................................................. 65 Materials and methods ............................................................................................................................ 69 Results...................................................................................................................................................... 73 Discussion ................................................................................................................................................ 79 Figures ..................................................................................................................................................... 83 Supplementary materials. ........................................................................................................................ 90 References ................................................................................................................................................ 95 CHAPTER VII ............................................................................................................................................ 98 Sentinel macrophages of zebrafish neuromasts ........................................................................................... 98 Introduction ........................................................................................................................................... 100 Materials and methods .......................................................................................................................... 102 Results.................................................................................................................................................... 105 Figures ................................................................................................................................................... 111 References .............................................................................................................................................. 119 General discussion ....................................................................................................................................... 121 Zebrafish as a model to study the immune system: advantages and pitfalls ......................................... 122 Type I IFN signalling is multilayered – insights from the zebrafish model about non-canonical modulators ............................................................................................................................................. 126 Regionalization of immune response .................................................................................................... 130 Leukocytes specialized in sentinel/homeostatic functions of particular sites ........................................ 132 Conclusions 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