title: Biology of Zika Virus Infection in Human Skin Cells.
abstract: Zika virus (ZIKV) is an emerging arbovirus of the Flaviviridae family, which includes dengue, West Nile, yellow fever, and Japanese encephalitis viruses, that causes a mosquito-borne disease transmitted by the Aedes genus, with recent outbreaks in the South Pacific. Here we examine the importance of human skin in the entry of ZIKV and its contribution to the induction of antiviral immune responses. We show that human dermal fibroblasts, epidermal keratinocytes, and immature dendritic cells are permissive to the most recent ZIKV isolate, responsible for the epidemic in French Polynesia. Several entry and/or adhesion factors, including DC-SIGN, AXL, Tyro3, and, to a lesser extent, TIM-1, permitted ZIKV entry, with a major role for the TAM receptor AXL. The ZIKV permissiveness of human skin fibroblasts was confirmed by the use of a neutralizing antibody and specific RNA silencing. ZIKV induced the transcription of Toll-like receptor 3 (TLR3), RIG-I, and MDA5, as well as several interferon-stimulated genes, including OAS2, ISG15, and MX1, characterized by strongly enhanced beta interferon gene expression. ZIKV was found to be sensitive to the antiviral effects of both type I and type II interferons. Finally, infection of skin fibroblasts resulted in the formation of autophagosomes, whose presence was associated with enhanced viral replication, as shown by the use of Torin 1, a chemical inducer of autophagy, and the specific autophagy inhibitor 3-methyladenine. The results presented herein permit us to gain further insight into the biology of ZIKV and to devise strategies aiming to interfere with the pathology caused by this emerging flavivirus.
  
  
  OBJECTIVE
  Zika virus (ZIKV) is an arbovirus belonging to the Flaviviridae family. Vector-mediated transmission of ZIKV is initiated when a blood-feeding female Aedes mosquito injects the virus into the skin of its mammalian host, followed by infection of permissive cells via specific receptors. Indeed, skin immune cells, including dermal fibroblasts, epidermal keratinocytes, and immature dendritic cells, were all found to be permissive to ZIKV infection. The results also show a major role for the phosphatidylserine receptor AXL as a ZIKV entry receptor and for cellular autophagy in enhancing ZIKV replication in permissive cells. ZIKV replication leads to activation of an antiviral innate immune response and the production of type I interferons in infected cells. Taken together, these results provide the first general insights into the interaction between ZIKV and its mammalian host.
authors.affiliation: Laboratoire MIVEGEC, UMR 224 IRD/CNRS/UM, Montpellier, France.
authors.forename: Rodolphe
authors.initials: R
authors.lastname: Hamel
authors.affiliation: INSERM, U944, Laboratoire de Pathologie et Virologie Moléculaire, Paris, France.
authors.forename: Ophélie
authors.initials: O
authors.lastname: Dejarnac
authors.affiliation: Laboratoire MIVEGEC, UMR 224 IRD/CNRS/UM, Montpellier, France.
authors.forename: Sineewanlaya
authors.initials: S
authors.lastname: Wichit
authors.affiliation: Laboratoire MIVEGEC, UMR 224 IRD/CNRS/UM, Montpellier, France.
authors.forename: Peeraya
authors.initials: P
authors.lastname: Ekchariyawat
authors.affiliation: Centre d'Étude d'Agents Pathogènes et Biotechnologies pour la Santé, CNRS-UMR 5236/UM, Montpellier, France.
authors.forename: Aymeric
authors.initials: A
authors.lastname: Neyret
authors.affiliation: Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
authors.forename: Natthanej
authors.initials: N
authors.lastname: Luplertlop
authors.affiliation: Laboratoire MIVEGEC, UMR 224 IRD/CNRS/UM, Montpellier, France.
authors.forename: Manuel
authors.initials: M
authors.lastname: Perera-Lecoin
authors.affiliation: Pathology Department, Prince of Songkla University, Songkla, Thailand.
authors.forename: Pornapat
authors.initials: P
authors.lastname: Surasombatpattana
authors.affiliation: Laboratoire MIVEGEC, UMR 224 IRD/CNRS/UM, Montpellier, France.
authors.forename: Loïc
authors.initials: L
authors.lastname: Talignani
authors.affiliation: Laboratoire MIVEGEC, UMR 224 IRD/CNRS/UM, Montpellier, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Thomas
authors.affiliation: Institut Louis Malardé, Papeete, Tahiti, French Polynesia.
authors.forename: Van-Mai
authors.initials: VM
authors.lastname: Cao-Lormeau
authors.affiliation: Environment and Infectious Risks Unit, Institut Pasteur, Paris, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Choumet
authors.affiliation: Centre d'Étude d'Agents Pathogènes et Biotechnologies pour la Santé, CNRS-UMR 5236/UM, Montpellier, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Briant
authors.affiliation: Département Infections et Epidémiologie, Institut Pasteur, Paris, France, and UMR PIMIT (I2T Team), Université de La Réunion, INSERM U1187, CNRS 9192, IRD 249, GIP-CYROI, Saint Clotilde, La Réunion, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Desprès
authors.affiliation: INSERM, U944, Laboratoire de Pathologie et Virologie Moléculaire, Paris, France.
authors.forename: Ali
authors.initials: A
authors.lastname: Amara
authors.affiliation: Centre d'Immunologie et des Maladies Infectieuses, INSERM, U1135, Sorbonne Universités, UPMC, APHP Hôpital Pitié-Salpêtrière, Paris, France.
authors.forename: Hans
authors.initials: H
authors.lastname: Yssel
authors.affiliation: Laboratoire MIVEGEC, UMR 224 IRD/CNRS/UM, Montpellier, France dorothee.misse@ird.fr.
authors.forename: Dorothée
authors.initials: D
authors.lastname: Missé
mesh_terms: D000330:Aedes / Q000821:virology; D000818:Animals; D001343:Autophagy / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D016207:Cytokines / Q000096:biosynthesis; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology / Q000821:virology*; D005347:Fibroblasts / Q000821:virology; D018067:Flaviviridae / Q000276:immunology / Q000502:physiology*; D018178:Flaviviridae Infections / Q000276:immunology / Q000821:virology; D057809:HEK293 Cells; D000072596:Hepatitis A Virus Cellular Receptor 1; D006801:Humans; D007303:Insect Vectors / Q000821:virology; D000072640:Interferon-Induced Helicase, IFIH1; D016899:Interferon-beta / Q000096:biosynthesis / Q000276:immunology; D015603:Keratinocytes / Q000821:virology*; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D064547:Myxovirus Resistance Proteins / Q000096:biosynthesis; D010588:Phagosomes / Q000276:immunology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D011971:Receptors, Immunologic; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D012867:Skin / Q000276:immunology / Q000821:virology; D051196:Toll-Like Receptor 3 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D051199:Toll-Like Receptor 7 / Q000276:immunology; D014452:Ubiquitins / Q000096:biosynthesis; D014709:Vero Cells; D053586:Virus Internalization; D014779:Virus Replication; D000094963:Axl Receptor Tyrosine Kinase; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D015815:Cell Adhesion Molecules; D016207:Cytokines; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases; D000072596:Hepatitis A Virus Cellular Receptor 1; D000072640:Interferon-Induced Helicase, IFIH1; D016899:Interferon-beta; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D064547:Myxovirus Resistance Proteins; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D020794:Receptor Protein-Tyrosine Kinases; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D011991:Receptors, Virus; D051196:Toll-Like Receptor 3; D051199:Toll-Like Receptor 7; D014452:Ubiquitins; D000094963:Axl Receptor Tyrosine Kinase; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C491143:HAVCR1 protein, human; C579071:MX1 protein, human; C495344:TLR3 protein, human; C495348:TLR7 protein, human; C074034:ISG15 protein, human; C084944:TYRO3 protein, human; C452820:RIGI protein, human; C452815:IFIH1 protein, human