title: Temperature-sensitive mutants of enterovirus 71 show attenuation in cynomolgus monkeys.
abstract: Enterovirus 71 (EV71) is one of the major causative agents of hand, foot and mouth disease and is sometimes associated with serious neurological disorders. In this study, an attempt was made to identify molecular determinants of EV71 attenuation of neurovirulence in a monkey infection model. An infectious cDNA clone of the virulent strain of EV71 prototype BrCr was constructed; temperature-sensitive (ts) mutations of an attenuated strain of EV71 or of poliovirus (PV) Sabin vaccine strains were then introduced into the infectious clone. In vitro and in vivo phenotypes of the parental and mutant viruses were analysed in cultured cells and in cynomolgus monkeys, respectively. Mutations in 3D polymerase (3D(pol)) and in the 3' non-translated region (NTR), corresponding to ts determinants of Sabin 1, conferred distinct temperature sensitivity to EV71. An EV71 mutant [EV71(S1-3')] carrying mutations in the 5' NTR, 3D(pol) and in the 3' NTR showed attenuated neurovirulence, resulting in limited spread of virus in the central nervous system of monkeys. These results indicate that EV71 and PV1 share common genetic determinants of neurovirulence in monkeys, despite the distinct properties in their original pathogenesis.
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
authors.forename: Minetaro
authors.initials: M
authors.lastname: Arita
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Shimizu
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
authors.forename: Noriyo
authors.initials: N
authors.lastname: Nagata
authors.affiliation: Division of Experimental Animals Research, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Ami
authors.affiliation: Division of Experimental Animals Research, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
authors.forename: Yuriko
authors.initials: Y
authors.lastname: Suzaki
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
authors.affiliation: Division of Clinical Investigation, Institute of Tropical Medicine, Nagasaki University, Nagasaki 852-8523, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Iwasaki
authors.affiliation: Department of Virology II, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama-shi, Tokyo 208-0011, Japan.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Miyamura
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics / Q000502:physiology; D020121:5' Untranslated Regions / Q000235:genetics / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000821:virology; D002478:Cells, Cultured; D020805:Central Nervous System Viral Diseases / Q000821:virology*; D004195:Disease Models, Animal; D004770:Enterovirus / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity*; D004769:Enterovirus Infections / Q000821:virology*; D005260:Female; D008252:Macaca fascicularis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D017955:Poliovirus / Q000235:genetics / Q000502:physiology; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000502:physiology; D013116:Spinal Cord / Q000821:virology; D010948:Viral Plaque Assay; D014774:Virulence / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; C090443:polymerase 3Dpol, poliovirus; D012324:RNA-Dependent RNA Polymerase