Mutational events during the primary propagation and consecutive passages of hepatitis E virus strain JE03-1760F in cell culture

Felipe R. Lorenzo, Toshinori Tanaka, Hideyuki Takahashi, Koji Ichiyama, Yu Hoshino, Kentaro Yamada, Jun Inoue, Masaharu Takahashi, Hiroaki Okamoto

Research output: Contribution to journalArticlepeer-review

39 Citations (Scopus)

Abstract

We recently developed a cell culture system for hepatitis E virus (HEV) in PLC/PRF/5 cells, using a genotype 3 HEV (JE03-1760F strain). Thirteen generations of consecutive passages of culture supernatant were successfully carried out in PLC/PRF/5 cells, with the highest HEV load reaching 108 copies/ml in the culture medium. Based on continuous release of progenies into culture medium, 50% tissue culture infectivity doses were estimated to be 2.0 × 103 copies for wild-type JE03-1760F and 1.4 × 102 copies for p13 (progeny in the thirteenth passage). Earlier appearance and greater increase in the yield of progenies in the culture supernatant were evident in p13 compared with wild-type. The cell culture-produced variants in primary propagation (p0) and consecutive passages (p5 [fifth passage], p10 [tenth], and p13) differed from the wild-type virus by 1, 9, 18, and 19 nucleotides (nt), respectively, over the entire genome of 7226 nt, excluding the poly(A) tail. Three of five non-synonymous mutations in p13 were shared by a variant (fifth passage) in another series of passages of JE03-1760F. These results suggest that adaptation of HEV variants to growth in vitro is associated with a limited number of mutations similar to hepatitis A virus.

Original languageEnglish
Pages (from-to)86-96
Number of pages11
JournalVirus Research
Volume137
Issue number1
DOIs
Publication statusPublished - 2008 Oct 1
Externally publishedYes

Keywords

  • Cell culture
  • Entire genome
  • Hepatitis E virus
  • Mutation
  • RNA viruses

ASJC Scopus subject areas

  • Cancer Research
  • Virology
  • Infectious Diseases

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