Successful infection of BALB/c mice by a swine hepatitis E virus clone constructed with reverse genetics

Abstract

BackgroundHepatitis E virus (HEV) is a leading cause of hepatitis worldwide. However, its infection biology and pathogenesis remain largely elusive. Furthermore, no proven medication is available for treating hepatitis E. Robust experimental models are urgently required to advance the research of HEV infection. Because of the lacking of a sophisticated small animal model, this study aimed to establish a mouse model of HEV infection.MethodsWe constructed a full-length swine HEV cDNA clone of genotype 4 (named as pGEM-HEV) by reverse genetics approach. And we inoculated with HEV RNA in BALB/c mice to establish small animal model for HEV infection and pathogenesis studies.ResultsThe capped RNA transcripts of pGEM-HEV prepared in vitro were replication-competent in HepG2 cells. Importantly, BALB/c mice intravenously inoculated with RNA transcripts of pGEM-HEV developed an active infection as shown by shedding viruses in feces, detectable negative strand of HEV in the liver, spleen and kidney, and causing liver inflammation.ConclusionIn this study, we successfully established of BALB/c mice-based small animal model for HEV provides an opportunity to further understand HEV pathogenesis and to develop effective antiviral medications.

Inhibition of hepatitis E virus replication by zinc‐finger antiviral Protein synergizes with IFN‐β

“…BC025308 ) was polymerase chain reaction (PCR)‐amplified and introduced into the C‐term of pcDNA3.1 vector and confirmed by restriction digestion and sequencing (pcDNA3.1‐ZAP). The construction and infectivity of full‐length HEV infectious cDNA clones (HEV) were identified in our previous study 22 . HEV 5ʹ‐UTR (27 nt) deleted infectious cDNA clone (HEV‐Δ5ʹ) was constructed by overlap PCR and started from the ATG of ORF1.…”

“…The construction and infectivity of full-length HEV infectious cDNA clones (HEV) were identified in our previous study. 22 HRP-and TRITC-conjugated secondary antibodies were obtained from Calbiochem (USA) and Jackson (USA), respectively.…”

Animal Models for Hepatitis E Virus

“…Recently, the same group has also used regular Balb/c mice and infected them with a full-length swine HEV cDNA clone of genotype 4, constructed via reverse genetics. HEV RNA was detected in feces, serum, and tissues, such as liver, spleen, colon, and kidney [131].…”

Infectious hepatitis E virus excreted into the vagina

“…Reverse transcription was performed with a reverse transcriptase kit (AMV XL for real‐time polymerase chain reaction [RT‐PCR], Takara, Japan) in accordance with the manufacturer's directions. HEV ORF1 (348 nucleotides), ORF2 (775 nucleotides), and ORF3 (369 nucleotides) were amplified through nested RT‐PCR in accordance with our previous study 23 . All three simultaneously positive HEV fragments were defined as HEV RNA positive.…”

“…HEV ORF1 (348 nucleotides), ORF2 (775 nucleotides), and ORF3 (369 nucleotides) were amplified through nested RT-PCR in accordance with our previous study. 23 All three simultaneously positive HEV fragments were defined as HEV RNA positive.…”