title: Lentivirus-mediated RNA interference against Japanese encephalitis virus infection in vitro and in vivo.
abstract: Japanese encephalitis virus, a serious mosquito-borne flavivirus, causes acute encephalitis in humans and many animals, with a high fatality rate. RNA interference is a reasonable antiviral mechanism for target gene silencing. In this study, four lentiviral shRNAs (LV-E1, LV-E2, LV-NS3 and LV-NS4b) were constructed. The results showed that four recombinant lentiviruses suppressed JEV replication in vitro. Through treatment with LV-E1 or LV-E2, the TCID50 values were reduced by 10(3)-fold during 120h post-challenge; the relative expression of viral mRNA was <7% or 13% in mouse and human neuroblastoma cells. Lentiviral shRNAs displayed robust inhibitory activity in various cells and against different genotypes of JEV. In vivo, pre-treatments of LV-E1 or LV-E2 resulted in no viral particles being observed in suckling mice brain sections. For 21days of observation, 100% of mice were protected against lethal JEV injection by two pre-treatments with LV-E1 or LV-E2; the survival of the mice pre-challenged with lethal JEV was 88.3%/66.7% by treatment with LV-E1 or LV-E2. LV-E1 and LV-E2 suppressed the induction of inflammatory mediators effectively in neuroblastoma cells and mice. Lentiviral shRNAs significantly inhibit JEV infection for long-term in vitro and in vivo and effectively reduce the inflammatory response and relieve encephalitis symptoms, highlighting the feasibility of using lentivirus-mediated RNAi for potential therapy in JEV infection.
authors.affiliation: Key Laboratory of Animal Diseases Diagnosis and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Shen
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Liu
authors.affiliation: Institute of Animal Husbandry & Veterinary Science, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China.
authors.forename: Denian
authors.initials: D
authors.lastname: Miao
authors.affiliation: Key Laboratory of Animal Diseases Diagnosis and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Ruibing
authors.initials: R
authors.lastname: Cao
authors.affiliation: Key Laboratory of Animal Diseases Diagnosis and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Animal Diseases Diagnosis and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China. Electronic address: shentingnau@hotmail.com.
authors.forename: Puyan
authors.initials: P
authors.lastname: Chen
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000008:administration & dosage* / Q000494:pharmacology / Q000627:therapeutic use; D001921:Brain / Q000473:pathology / Q000821:virology; D004195:Disease Models, Animal; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000502:physiology*; D004672:Encephalitis, Japanese / Q000188:drug therapy / Q000473:pathology / Q000821:virology*; D005260:Female; D007249:Inflammation / Q000473:pathology; D016086:Lentivirus / Q000235:genetics*; D008807:Mice, Inbred BALB C; D034622:RNA Interference; D012367:RNA, Viral / Q000008:administration & dosage* / Q000494:pharmacology / Q000627:therapeutic use; D016019:Survival Analysis; D014762:Viral Interference; D019562:Viral Load; D014776:Virus Cultivation; D014779:Virus Replication
chemical_list: D000998:Antiviral Agents; D012367:RNA, Viral
keywords: Antiviral; In vitro and in vivo; Inflammatory response; JEV; Lentiviral shRNA