title: Genomic changes detected after serial passages in cell culture of virulent human G1P[8] rotaviruses.
abstract: Serial passages of a virulent mouse rotavirus in cell cultures caused a loss of virulence in mice. To gain insight into the genomic mutations in human rotavirus during cell culture and its attenuation in humans, we serially passaged three wild type human G1P[8] rotavirus strains (Wa, DC3695, DC5685) derived from diarrheal stool samples up to 60 times in two different cell cultures (human colon adenocarcinoma cell line: HT29, and primary African green monkey kidney cells: primary AGMK). We sequenced the whole genomes of 60 times-passaged strains and compared them with those of the original viruses. Most substitutions were detected in VP4, followed by substitutions in VP7 and NSP4 genes. Substitution at amino acid 385 in the putative VP4 fusion domain and substitution T45M in NSP4 genes were detected in all AGMK-passaged strains, respectively. These genomic changes are likely to correlate with a loss of rotavirus virulence in humans.
authors.affiliation: Department of Pediatrics, Sapporo Medical University School of Medicine, South-1 West-16, Chuo-ku, Sapporo 060-8543, Japan; Epidemiology Section, Laboratory of Infectious Disease, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Building 50, Room 6308, 50 South Drive, MSC 8026, Bethesda, MD 20892-8026, USA. Electronic address: tsugawat@sapmed.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tsugawa
authors.affiliation: Department of Pediatrics, Sapporo Medical University School of Medicine, South-1 West-16, Chuo-ku, Sapporo 060-8543, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tsutsumi
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002522:Chlorocebus aethiops; D016679:Genome, Viral / Q000235:genetics*; D023281:Genomics; D019073:HT29 Cells; D006801:Humans; D051379:Mice; D009154:Mutation / Q000235:genetics; D012401:Rotavirus / Q000235:genetics*; D012400:Rotavirus Infections / Q000821:virology; D012692:Serial Passage; D014709:Vero Cells; D014774:Virulence / Q000235:genetics*
keywords: Cell culture; Mutation; Rotavirus; Sequence; Serial passage; Virulence