title: Use of single-gene reassortant viruses to study the role of avian influenza A virus genes in attenuation of wild-type human influenza A virus for squirrel monkeys and adult human volunteers.
abstract: The transfer of six internal RNA segments from the avian influenza A/Mallard/New York/6750/78 (H2N2) virus reproducibly attenuates human influenza A viruses for squirrel monkeys and adult humans. To identify the avian influenza A virus genes that specify the attenuation and host range restriction of avian-human (ah) influenza A reassortant viruses (referred to as ah reassortants), we isolated six single-gene reassortant viruses (SGRs), each having a single internal RNA segment of the influenza A/Mallard/New York/6750/78 virus and seven RNA segments from the human influenza A/Los Angeles/2/87 (H3N2) wild-type virus. To assess the level of attenuation, we compared each SGR with the A/Los Angeles/2/87 wild-type virus and a 6-2 gene ah reassortant (having six internal RNA segments from the avian influenza A virus parent and two genes encoding the hemagglutinin and neuraminidase glycoproteins from the wild-type human influenza A virus) for the ability to replicate in seronegative squirrel monkeys and adult human volunteers. In monkeys and humans, replication of the 6-2 gene ah reassortant was highly restricted. In humans, the NS, M, PB2, and PB1 SGRs each replicated significantly less efficiently (P less than 0.05) than the wild-type human influenza A virus parent, suggesting that each of these genes contributes to the attenuation phenotype. In monkeys, only the NP, PB2, and possibly the M genes contributed to the attenuation phenotype. These discordant observations, particularly with regard to the NP SGR, indicate that not all genetic determinants of attenuation of influenza A viruses for humans can be identified during studies of SGRs conducted with monkeys. The PB2 and M SGRs that were attenuated in humans each exhibited a new phenotype that was not observed for either parental virus. Thus, it was not possible to determine whether avian influenza virus PB2 or M gene itself or a specific constellation of avian and human influenza A virus specified restriction of virus replication in humans.
authors.affiliation: Department of International Health, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland.
authors.forename: M L
authors.initials: ML
authors.lastname: Clements
authors.forename: E K
authors.initials: EK
authors.lastname: Subbarao
authors.forename: L F
authors.initials: LF
authors.lastname: Fries
authors.forename: R A
authors.initials: RA
authors.lastname: Karron
authors.forename: W T
authors.initials: WT
authors.lastname: London
authors.forename: B R
authors.initials: BR
authors.lastname: Murphy
mesh_terms: D000328:Adult; D000818:Animals; D001483:Base Sequence; D005814:Genes, Viral; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D007252:Influenza Vaccines / Q000302:isolation & purification; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics; D012453:Saimiri; D014162:Transfection; D014613:Vaccines, Attenuated / Q000302:isolation & purification; D014774:Virulence / Q000235:genetics; D014779:Virus Replication / Q000235:genetics
chemical_list: D007252:Influenza Vaccines; D012367:RNA, Viral; D014613:Vaccines, Attenuated