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Cited by 1,523Influenza Virus Research StudiesVirology and Viral DiseasesViral Infections and Outbreaks Research

Experimental adaptation of an influenza H5 HA confers respiratory droplet transmission to a reassortant H5 HA/H1N1 virus in ferrets

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Imai, Masaki, Watanabe, Tokiko, Hatta, Masato, Das, Subash C., Ozawa, Makoto, Shinya, Kyoko, Zhong, Gongxun, Hanson, Anthony +10 more·2012·Nature
DOI 10.1038/nature10831PMC PMC3388103
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Abstract

Only four mutations in H5N1 HA are required to enable ferret-to-ferret transmission of a reassortant virus containing the H5 HA and the remaining seven gene segments from a human pandemic H1N1 influenza virus. Whether avian H5N1 viruses can gain the ability to transmit between humans was uncertain. The viral haemagglutinin protein (HA) mediates virus binding to host-specific cellular receptors, but previous studies have shown that alterations in HA that enable binding to human-type receptors are not sufficient to enable respiratory droplet transmission of H5N1 viruses in ferrets, the best animal model for human-to-human transmission. Imai et al. show that only four mutations in H5N1 HA are required to enable ferret-to-ferret transmission of a reassortant virus containing H5 HA, with the remaining genes from human pandemic H1N1 influenza virus. It is probable that further adaptations in other avian virus genes would be required to mediate transmission of wholly avian H5N1 in mammals, but human H1N1 and H5N1 viruses are genetically compatible and the emergence of H5-HA-containing viruses might be expected to cause a pandemic because humans lack immunity to H5 viruses. Knowledge of the mutations involved in adapting H5 HA to mammalian transmission could help with surveillance and monitoring of H5N1 viruses adapting towards pandemic potential. Highly pathogenic avian H5N1 influenza A viruses occasionally infect humans, but currently do not transmit efficiently among humans. The viral haemagglutinin (HA) protein is a known host-range determinant as it mediates virus binding to host-specific cellular receptors1,2,3. Here we assess the molecular changes in HA that would allow a virus possessing subtype H5 HA to be transmissible among mammals. We identified a reassortant H5 HA/H1N1 virus—comprising H5 HA (from an H5N1 virus) with four mutations and the remaining seven gene segments from a 2009 pandemic H1N1 virus—that was capable of droplet transmission in a ferret model. The transmissible H5 reassortant virus preferentially recognized human-type receptors, replicated efficiently in ferrets, caused lung lesions and weight loss, but was not highly pathogenic and did not cause mortality. These results indicate that H5 HA can convert to an HA that supports efficient viral transmission in mammals; however, we do not know whether the four mutations in the H5 HA identified here would render a wholly avian H5N1 virus transmissible. The genetic origin of the remaining seven viral gene segments may also critically contribute to transmissibility in mammals. Nevertheless, as H5N1 viruses continue to evolve and infect humans, receptor-binding variants of H5N1 viruses with pandemic potential, including avian–human reassortant viruses as tested here, may emerge. Our findings emphasize the need to prepare for potential pandemics caused by influenza viruses possessing H5 HA, and will help individuals conducting surveillance in regions with circulating H5N1 viruses to recognize key residues that predict the pandemic potential of isolates, which will inform the development, production and distribution of effective countermeasures.

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  • Transmission-enhanced H5N1 avian influenza can spread through the air from ferret to ferret, but its relative contagiousness is questionable due to small sample sizes.

    “Transmission-enhanced H5N1 avian influenza can spread through the air from ferret to ferret, but its relative contagiousness is questionable due to small sample sizes -and of course, ferrets are not humans 4,5 .”

    From Human innovation and the creative agency of the world in the age of generative AI (2024) · cites this paper as [5]

    The transmission-results passage (labeled Abstract; no page given) reports airborne respiratory-droplet transmission to contact ferrets, with virus recovered from 4 of 6, and shows that the comparison groups were also small. But the tested virus was a mutant H5 HA/H1N1 reassortant, not a wholly avian H5N1 virus; the paper explicitly says it does not know whether the mutations would make a wholly avian H5N1 virus transmissible. It does not explicitly attribute uncertainty about relative contagiousness to sample size. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • The high likelihood that newly identified pandemic-class agents will be publicly described, including novel versions devised by future LLMs, underscores the importance of verifying that screening is conducted against an up-to-date database.

    “The high likelihood that newly identified pandemic-class agents will be publicly described 4,5,26,27 , including novel versions devised by future LLMs, underscore the importance of verifying that screening is conducted against an up-to-date database.”

    From Human innovation and the creative agency of the world in the age of generative AI (2024) · cites this paper as [5]

    The Abstract publicly describes an experimentally adapted virus with pandemic potential and says the authors strongly endorse sharing their data globally. This supports the paper as an example of public description, but it does not establish a high likelihood that newly identified pandemic-class agents generally will be publicly described. The LLM and screening-database points are the citing authors’ extension, not findings of this source. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 26

  1. [1]Glaser L A single amino acid substitution in 1918 influenza virus hemagglutinin changes receptor binding specificity J Virol 79 11533 11536 2005 16103207 PMC1193621
  2. [2]Matrosovich M Early alterations of the receptor-binding properties of H1, H2, and H3 avian influenza virus hemagglutinins after their introduction into mammals J Virol 74 8502 8512 2000 10954551 PMC116362
  3. [3]Rogers GN Paulson JC Receptor determinants of human and animal influenza virus isolates: differences in receptor specificity of the H3 hemagglutinin based on species of origin Virology 127 361 373 1983 6868370
  4. [4]Auewarakul P An avian influenza H5N1 virus that binds to a human-type receptor J Virol 81 9950 9955 2007 17626098 PMC2045398
  5. [5]Gambaryan A Evolution of the receptor binding phenotype of influenza A (H5) viruses Virology 344 432 438 2006 16226289
  6. [6]Stevens J Recent avian H5N1 viruses exhibit increased propensity for acquiring human receptor specificity Journal of molecular biology 381 1382 1394 2008 18672252 PMC2519951
  7. [7]Wang W Glycosylation at 158N of the hemagglutinin protein and receptor binding specificity synergistically affect the antigenicity and immunogenicity of a live attenuated H5N1 A/Vietnam/1203/2004 vaccine virus in ferrets J Virol 84 6570 6577 2010 20427525 PMC2903256
  8. [8]Watanabe Y Acquisition of human-type receptor binding specificity by new H5N1 influenza virus sublineages during their emergence in birds in Egypt PLoS pathogens 7 e1002068 2011 21637809 PMC3102706
  9. [9]Yamada S Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors Nature 444 378 382 2006 17108965
  10. [10]Jackson S Reassortment between avian H5N1 and human H3N2 influenza viruses in ferrets: a public health risk assessment J Virol 83 8131 8140 2009 19493997 PMC2715755
  11. [11]Maines TR Lack of transmission of H5N1 avian-human reassortant influenza viruses in a ferret model Proceedings of the National Academy of Sciences of the United States of America 103 12121 12126 2006 16880383 PMC1567706
  12. [12]Maines TR Effect of receptor binding domain mutations on receptor binding and transmissibility of avian influenza H5N1 viruses Virology 413 139 147 2011 21397290 PMC5470842
  13. [13]Yen HL Inefficient transmission of H5N1 influenza viruses in a ferret contact model J Virol 81 6890 6898 2007 17459930 PMC1933302
  14. [14]Chen LM In vitro evolution of H5N1 avian influenza virus toward human-type receptor specificity Virology 422 105 113 2012 22056389 PMC5480292
  15. [15]Cline TD Increased pathogenicity of a reassortant 2009 pandemic H1N1 influenza virus containing an H5N1 hemagglutinin J Virol 85 12262 12270 2011 21917948 PMC3209346
  16. [16]Octaviani CP Ozawa M Yamada S Goto H Kawaoka Y High level of genetic compatibility between swine-origin H1N1 and highly pathogenic avian H5N1 influenza viruses J Virol 84 10918 10922 2010 20686037 PMC2950597
  17. [17]Itoh Y In vitro and in vivo characterization of new swine-origin H1N1 influenza viruses Nature 460 1021 1025 2009 19672242 PMC2748827
  18. [18]Shinya K Avian flu: influenza virus receptors in the human airway Nature 440 435 436 2006 16554799
  19. [20]Bussey KA Bousse TL Desmet EA Kim B Takimoto T PB2 residue 271 plays a key role in enhanced polymerase activity of influenza A viruses in mammalian host cells J Virol 84 4395 4406 2010 20181719 PMC2863787
  20. [21]Hatta M Gao P Halfmann P Kawaoka Y Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses Science 293 1840 1842 2001 11546875
  21. [22]Li Z Molecular basis of replication of duck H5N1 influenza viruses in a mammalian mouse model J Virol 79 12058 12064 2005 16140781 PMC1212590
  22. [23]Mehle A Doudna JA Adaptive strategies of the influenza virus polymerase for replication in humans Proceedings of the National Academy of Sciences of the United States of America 106 21312 21316 2009 19995968 PMC2789757
  23. [24]Yamada S Biological and structural characterization of a host-adapting amino acid in influenza virus PLoS pathogens 6 e1001034 2010 20700447 PMC2916879
  24. [25]Salzberg SL Genome analysis linking recent European and African influenza (H5N1) viruses Emerging infectious diseases 13 713 718 2007 17553249 PMC2432181
  25. [26]Treanor JJ Campbell JD Zangwill KM Rowe T Wolff M Safety and immunogenicity of an inactivated subvirion influenza A (H5N1) vaccine The New England journal of medicine 354 1343 1351 2006 16571878
  26. [27]Neumann G Generation of influenza A viruses entirely from cloned cDNAs Proceedings of the National Academy of Sciences of the United States of America 96 9345 9350 1999 10430945 PMC17785

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Authors
Imai, M., Watanabe, T., Hatta, M., Das, S. C., Ozawa, M., Shinya, K., Zhong, G., Hanson, A., Katsura, H., Watanabe, S., Li, C., Kawakami, E., Yamada, S., Kiso, M., Suzuki, Y., Maher, E. A., Neumann, G., & Kawaoka, Y.
Affiliations
  • University of Wisconsin–Madison (US)
  • Kobe University (JP)
  • Institute of Medical Science, The University of Tokyo (JP)
  • Chubu University (JP)
Corresponding author
Kawaoka, Yoshihiro
ORCID
Venue
Nature
Publisher
Springer Science and Business Media LLC
Published
2012-05-02
Volume
486
Issue
7403
Pages
420-428
ISSN
  • 0028-0836 (ISSN-L)
  • 1476-4687
DOI
10.1038/nature10831
PMC
PMC3388103
OpenAlex
W2112170460
Cited by
1,523
References
3
Citations by year
2026: 49 · 2025: 74 · 2024: 103 · 2023: 70 · 2022: 63 · 2021: 73 · 2020: 78 · 2019: 73 · 2018: 81 · 2017: 95 · 2016: 90 · 2015: 136 · 2014: 207 · 2013: 246 · 2012: 85
FWCI
161.40
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
Funders
  • Bill and Melinda Gates Foundation (OPPGH5383)
  • National Institutes of Health
  • Centers for Disease Control and Prevention
  • Japan Society for the Promotion of Science (24658245, 20390028)
  • Japan Science and Technology Agency
  • National Institute of Allergy and Infectious Diseases (R01AI069274, T32AI078985)
  • Exploratory Research for Advanced Technology
Topics
  • Health Sciences › Medicine › Epidemiology › Influenza Virus Research Studies
  • Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
  • Health Sciences › Medicine › Infectious Diseases › Viral Infections and Outbreaks Research
Keywords
respiratory droplet transmission, reassortant viruses, host range adaptation, ferret model, pandemic potential, avian-to-human transmission, lung lesions, 2009 H1N1 pandemic
MeSH terms
Adaptation, Physiological, Animals, Birds, Body Fluids, Cell Line, Dogs, Female, Ferrets, Influenza in Birds, Hot Temperature, HeLa Cells, Humans, Influenza, Human, Orthomyxoviridae Infections, Population Surveillance, Receptors, Virus, Respiratory System, Security Measures, Zoonoses, Reassortant Viruses, Molecular Epidemiology, Evolution, Molecular, Hemagglutinin Glycoproteins, Influenza Virus, Bioterrorism, Influenza A Virus, H1N1 Subtype, Influenza A Virus, H5N1 Subtype, Protein Stability, HEK293 Cells, Pandemics

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