Rescue of a genotype 4 human hepatitis E virus from cloned cDNA and characterization of intergenotypic chimeric viruses in cultured human liver cells and in pigs

Laura Córdoba, Alicia R Feagins, Tanja Opriessnig, Caitlin M Cossaboom, Barbara A Dryman, Yao-Wei Huang, Xiang-Jin Meng

Research output: Contribution to journalArticlepeer-review

Abstract

Hepatitis E virus (HEV) is an important but extremely understudied human pathogen. Genotypes 1 and 2 are restricted to humans, whereas genotypes 3 and 4 are zoonotic, infecting both humans and pigs. This report describes, for the first time, the successful rescue of infectious HEV in vitro and in vivo from cloned cDNA of a genotype 4 human HEV (strain TW6196E). The complete genomic sequence of the TW6196E virus was determined and a full-length cDNA clone (pHEV-4TW) was assembled. Capped RNA transcripts from the pHEV-4TW clone were replication competent in Huh7 cells and infectious in HepG2/C3A cells. Pigs inoculated intrahepatically with capped RNA transcripts from pHEV-4TW developed an active infection, as evidenced by faecal virus shedding and seroconversion, indicating the successful rescue of infectious genotype 4 HEV and cross-species infection of pigs by a genotype 4 human HEV. To demonstrate the utility of the genotype 4 HEV infectious clone and to evaluate the potential viral determinant(s) for species tropism, four intergenotypic chimeric clones were constructed by swapping various genomic regions between genotypes 1 and 4, and genotypes 1 and 3. All four chimeric clones were replication competent in Huh7 cells, but only the two chimeras with sequences swapped between genotypes 1 and 4 human HEVs produced viruses capable of infecting HepG2/C3A cells. None of the four chimeras was able to establish a robust infection in pigs. The availability of a genotype 4 HEV infectious clone affords an opportunity to delineate the molecular mechanisms of HEV cross-species infection in the future.
Original languageEnglish
Pages (from-to)2183-94
Number of pages12
JournalJournal of General Virology
Volume93
Issue numberPt 10
DOIs
Publication statusPublished - Oct 2012

Keywords

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line, Tumor
  • Cloning, Molecular
  • DNA, Complementary
  • Feces
  • Genome, Viral
  • Genotype
  • Hep G2 Cells
  • Hepatitis E
  • Hepatitis E virus
  • Humans
  • Liver
  • Molecular Sequence Data
  • RNA Caps
  • RNA, Viral
  • Sequence Analysis
  • Swine
  • Virus Replication
  • Virus Shedding

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