Construction and characterization of an infectious cDNA clone of rat hepatitis E virus

Li, Tiancheng; Yang, Tingting; Yoshizaki, Sayaka; Ami, Yasushi; Suzaki, Yuriko; Ishii, Koji; Haga, Kei; Nakamura, Tomofumi; Ochiai, Susumu; Wakita, Takaji; Johne, Reimar

doi:10.1099/vir.0.000072

Cited by 40 publications

(36 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Contrary to the cDNA clone of this previous study, which contains 23 mutations compared to the original sequence, pLA-B350 is based on the consensus sequence of the LA-B350 strain and any PCR-induced mutations were corrected. LA-B350 also has the advantage of replicating in the commonly used Huh7 and HepG2/C3A hepatoma cell lines, but we could not detect solid replication in PLC/PRF/5 cells, contrary to the results for R63/DEU/2009 (Li et al, 2015). …”

Section: Discussioncontrasting

confidence: 99%

See 1 more Smart Citation

A rat model for hepatitis E virus

Debing

Mishra

Verbeken

et al. 2016

Disease Models &Amp; Mechanisms

View full text Add to dashboard Cite

Hepatitis E virus (HEV) is one of the prime causes of acute viral hepatitis, and chronic hepatitis E is increasingly recognized as an important problem in the transplant setting. Nevertheless, the fundamental understanding of the biology of HEV replication is limited and there are few therapeutic options. The development of such therapies is partially hindered by the lack of a robust and convenient animal model. We propose the infection of athymic nude rats with the rat HEV strain LA-B350 as such a model. A cDNA clone, pLA-B350, was constructed and the infectivity of its capped RNA transcripts was confirmed in vitro and in vivo. Furthermore, a subgenomic replicon, pLA-B350/luc, was constructed and validated for in vitro antiviral studies. Interestingly, rat HEV proved to be less sensitive to the antiviral activity of α-interferon, ribavirin and mycophenolic acid than genotype 3 HEV (a strain that infects humans). As a proof-of-concept, part of the C-terminal polymerase sequence of pLA-B350/luc was swapped with its genotype 3 HEV counterpart: the resulting chimeric replicon replicated with comparable efficiency as the wild-type construct, confirming that LA-B350 strain is amenable to humanization (replacement of certain sequences or motifs by their counterparts from human HEV strains). Finally, ribavirin effectively inhibited LA-B350 replication in athymic nude rats, confirming the suitability of the rat model for antiviral studies.

show abstract

Section: Discussioncontrasting

confidence: 99%

“…The construction of a cDNA clone of rat HEV strain R63/DEU/2009 was recently reported (Li et al, 2015). Contrary to the cDNA clone of this previous study, which contains 23 mutations compared to the original sequence, pLA-B350 is based on the consensus sequence of the LA-B350 strain and any PCR-induced mutations were corrected.…”

Section: Discussionmentioning

confidence: 99%

A rat model for hepatitis E virus

Debing

Mishra

Verbeken

et al. 2016

Disease Models &Amp; Mechanisms

View full text Add to dashboard Cite

show abstract

“…The nucleotide sequencing was carried out using an ABI 3130 Genetic Analyzer automated sequencer (Applied Biosystems, Foster City, CA). The sequences of entire genomes were confirmed by next‐generation sequence analysis as described previously (Li et al., ).…”

Section: Methodsmentioning

confidence: 99%

Characterization of porcine sapelovirus isolated from Japanese swine with PLC/PRF/5 cells

Bai

Liu

Fang³

et al. 2017

Transbound Emerg Dis

View full text Add to dashboard Cite

Porcine sapelovirus (PSV) is a causative agent of neurological disorders, fertility disorders and dermal lesions of swine. In this study, we isolated two PSV strains, Jpsv477 and Jpsv1315, from swine faecal specimens using a PLC/PRF/5 cell culture system. The PSV infection of PLC/PRF/5 cells induced a cytopathic effect (CPE). Two types of virus particles with identical diameter (~35 nm) but different densities (1.300 and 1.285 g/cm ) were observed in the cell culture supernatants. Analysis of the entire genome sequence of Jpsv477 and Jpsv1315 revealed that both strains possess 7,558 nucleotides and the poly (A) tail and have a typical PSV genome organization consisting of a 5' terminal untranslated region (5'UTR), a large open reading frame (ORF), and a 3' terminal untranslated region (3'UTR). The ORF encodes a single polyprotein that is subsequently processed into a leader protein (L), four structural proteins (VP1, VP2, VP3 and VP4) and seven functional proteins (2A, 2B, 2C, 3A, 3B, 3C and 3D). The structural proteins VP1, VP2, VP3 and VP4 have molecular masses of ~35, ~26, ~25 and ~6 kDa. The N-terminal amino acid sequence analysis of VP1, VP2, VP3 and VP4 confirmed that the cleavage sites between VP4 and VP2, VP2 and VP3, and VP3 and VP1 are K/A, Q/G and Q/G, respectively. We further confirmed that HepG2/C3A, Vero E6 and primary green monkey kidney cells (PGMKC) were also susceptible to PSV infection. The stability assay demonstrated that PSV was inactivated by heating at 60°C for 10 min or 65°C for 5 min. The virus also lost infectivity by incubation with 62.5 ppm of NaClO for 30 min.

show abstract

“…However, in two other reports, the injection of HEV-1, -2, -3 (swine) and -4 (wild boar) failed to induce an efficient infection in Sprague-Dawley or Wistar rats [101,103]. Rats were also inoculated with different RNA transcripts from infectious cDNA clones of rat HEV [104], HEV-4 [105] and swine HEV-3 [106] leading to successful infections. Moreover, attempts to infect Wistar and nude rats with ferret HEV failed [78].…”

Section: Animal Models Of Hevmentioning

confidence: 99%

Zoonotic Hepatitis E Virus: Classiﬁcation, Animal Reservoirs and Transmission Routes

Doceul

Bagdassarian

Demange

et al. 2016

Viruses

208

199

View full text Add to dashboard Cite

During the past ten years, several new hepatitis E viruses (HEVs) have been identified in various animal species. In parallel, the number of reports of autochthonous hepatitis E in Western countries has increased as well, raising the question of what role these possible animal reservoirs play in human infections. The aim of this review is to present the recent discoveries of animal HEVs and their classification within the Hepeviridae family, their zoonotic and species barrier crossing potential, and possible use as models to study hepatitis E pathogenesis. Lastly, this review describes the transmission pathways identified from animal sources.

show abstract

Construction and characterization of an infectious cDNA clone of rat hepatitis E virus

Cited by 40 publications

References 28 publications

A rat model for hepatitis E virus

A rat model for hepatitis E virus

Characterization of porcine sapelovirus isolated from Japanese swine with PLC/PRF/5 cells

Zoonotic Hepatitis E Virus: Classiﬁcation, Animal Reservoirs and Transmission Routes

Contact Info

Product

Resources

About