Subjects

Abstract

Avian influenza A viruses rarely infect humans; however, when human infection and subsequent human-to-human transmission occurs, worldwide outbreaks (pandemics) can result. The recent sporadic infections of humans in China with a previously unrecognized avian influenza A virus of the H7N9 subtype (A(H7N9)) have caused concern owing to the appreciable case fatality rate associated with these infections (more than 25%), potential instances of human-to-human transmission1, and the lack of pre-existing immunity among humans to viruses of this subtype. Here we characterize two early human A(H7N9) isolates, A/Anhui/1/2013 (H7N9) and A/Shanghai/1/2013 (H7N9); hereafter referred to as Anhui/1 and Shanghai/1, respectively. In mice, Anhui/1 and Shanghai/1 were more pathogenic than a control avian H7N9 virus (A/duck/Gunma/466/2011 (H7N9); Dk/GM466) and a representative pandemic 2009 H1N1 virus (A/California/4/2009 (H1N1pdm09); CA04). Anhui/1, Shanghai/1 and Dk/GM466 replicated well in the nasal turbinates of ferrets. In nonhuman primates, Anhui/1 and Dk/GM466 replicated efficiently in the upper and lower respiratory tracts, whereas the replicative ability of conventional human influenza viruses is typically restricted to the upper respiratory tract of infected primates. By contrast, Anhui/1 did not replicate well in miniature pigs after intranasal inoculation. Critically, Anhui/1 transmitted through respiratory droplets in one of three pairs of ferrets. Glycan arrays showed that Anhui/1, Shanghai/1 and A/Hangzhou/1/2013 (H7N9) (a third human A(H7N9) virus tested in this assay) bind to human virus-type receptors, a property that may be critical for virus transmissibility in ferrets. Anhui/1 was found to be less sensitive in mice to neuraminidase inhibitors than a pandemic H1N1 2009 virus, although both viruses were equally susceptible to an experimental antiviral polymerase inhibitor. The robust replicative ability in mice, ferrets and nonhuman primates and the limited transmissibility in ferrets of Anhui/1 suggest that A(H7N9) viruses have pandemic potential.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    China–WHO Joint Mission on Human Infection with Avian Influenza A(H7N9) Virus. 18–24 April 2013, Mission Report. (2013)

  2. 2.

    et al. Human infection with a novel avian-origin influenza A (H7N9) virus. N. Engl. J. Med. 368, 1888–1897 (2013)

  3. 3.

    Number of confirmed human cases of avian influenza A(H7N9) reported to WHO. Report 8 – data in WHO/HQ as of 30 May 2013, 15:45 GMT+1. (2013)

  4. 4.

    et al. Genetic analysis of novel avian A(H7N9) influenza viruses isolated from patients in China, February to April 2013. Euro Surveill. 18, 20453 (2013)

  5. 5.

    et al. Genomic signature and protein sequence analysis of a novel influenza A (H7N9) virus that causes an outbreak in humans in China. Microbes Infect. (2013)

  6. 6.

    et al. Single amino acid substitutions in influenza haemagglutinin change receptor binding specificity. Nature 304, 76–78 (1983)

  7. 7.

    , , & Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses. Science 293, 1840–1842 (2001)

  8. 8.

    , & Rescue of an influenza A virus wild-type PB2 gene and a mutant derivative bearing a site-specific temperature-sensitive and attenuating mutation. J. Virol. 67, 7223–7228 (1993)

  9. 9.

    et al. Molecular basis of replication of duck H5N1 influenza viruses in a mammalian mouse model. J. Virol. 79, 12058–12064 (2005)

  10. 10.

    et al. In vitro and in vivo characterization of new swine-origin H1N1 influenza viruses. Nature 460, 1021–1025 (2009)

  11. 11.

    et al. Severity of pneumonia due to new H1N1 influenza virus in ferrets is intermediate between that due to seasonal H1N1 virus and highly pathogenic avian influenza H5N1 virus. J. Infect. Dis. 201, 993–999 (2010)

  12. 12.

    et al. Classical swine H1N1 influenza viruses confer cross protection from swine-origin 2009 pandemic H1N1 influenza virus infection in mice and ferrets. Virology 408, 128–133 (2010)

  13. 13.

    et al. Transmission and pathogenesis of swine-origin 2009 A(H1N1) influenza viruses in ferrets and mice. Science 325, 484–487 (2009)

  14. 14.

    et al. Pathogenesis and transmission of swine-origin 2009 A(H1N1) influenza virus in ferrets. Science 325, 481–483 (2009)

  15. 15.

    et al. Experimental adaptation of an influenza H5 HA confers respiratory droplet transmission to a reassortant H5 HA/H1N1 virus in ferrets. Nature 486, 420–428 (2012)

  16. 16.

    et al. Reassortment between avian H5N1 and human H3N2 influenza viruses in ferrets: a public health risk assessment. J. Virol. 83, 8131–8140 (2009)

  17. 17.

    et al. Lack of transmission of H5N1 avian-human reassortant influenza viruses in a ferret model. Proc. Natl Acad. Sci. USA 103, 12121–12126 (2006)

  18. 18.

    et al. Airborne transmission of influenza A/H5N1 virus between ferrets. Science 336, 1534–1541 (2012)

  19. 19.

    et al. Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors. Nature 444, 378–382 (2006)

  20. 20.

    et al. In vitro assessment of attachment pattern and replication efficiency of H5N1 influenza A viruses with altered receptor specificity. J. Virol. 84, 6825–6833 (2010)

  21. 21.

    , , , & Quantitative characterization of glycan-receptor binding of H9N2 influenza A virus hemagglutinin. PLoS ONE 8, e59550 (2013)

  22. 22.

    et al. Glycan topology determines human adaptation of avian H5N1 virus hemagglutinin. Nature Biotechnol. 26, 107–113 (2008)

  23. 23.

    et al. In vitro evolution of H5N1 avian influenza virus toward human-type receptor specificity. Virology 422, 103–113 (2012)

  24. 24.

    et al. Contemporary North American influenza H7 viruses possess human receptor specificity: implications for virus transmissibility. Proc. Natl Acad. Sci. USA 105, 7558–7563 (2008)

  25. 25.

    et al. Mutations in a conserved residue in the influenza virus neuraminidase active site decreases sensitivity to Neu5Ac2en-derived inhibitors. J. Virol. 72, 2456–2462 (1998)

  26. 26.

    Emergence of avian influenza A(H7N9) virus causing severe human illness - China, February–April 2013. MMWR Morb. Mortal. Wkly Rep. 62, 366–371 (2013)

  27. 27.

    et al. Evaluation of neuraminidase enzyme assays using different substrates to measure susceptibility of influenza virus clinical isolates to neuraminidase inhibitors: report of the neuraminidase inhibitor susceptibility network. J. Clin. Microbiol. 41, 742–750 (2003)

  28. 28.

    et al. Poor responses to oseltamivir treatment in a patient with influenza A (H7N9) virus infection. Emerg. Microbes & Infections 2, e27 (2013)

  29. 29.

    , , , & Basal cells of differentiated bronchial epithelium are more susceptible to rhinovirus infection. Am. J. Respir. Cell Mol. Biol. 38, 517–523 (2008)

  30. 30.

    et al. Generation of influenza A viruses entirely from cloned cDNAs. Proc. Natl Acad. Sci. USA 96, 9345–9350 (1999)

  31. 31.

    , & Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108, 193–199 (1991)

  32. 32.

    , , , & Replication and transmission of influenza viruses in Japanese quail. Virology 310, 8–15 (2003)

  33. 33.

    et al. Highly pathogenic H5N1 influenza virus infection in migratory birds. Science 309, 1206 (2005)

  34. 34.

    et al. Infectivity, transmission, and pathology of human H7N9 influenza in ferrets and pigs. Science (2013)

  35. 35.

    et al. A recurring motif for antibody recognition of the receptor-binding site of influenza hemagglutinin. Nature Struct. Mol. Biol. 20, 363–370 (2013)

  36. 36.

    et al. Architecture of ribonucleoprotein complexes in influenza A virus particles. Nature 439, 490–492 (2006)

Download references

Acknowledgements

We thank Y. Shu for A/Anhui/1/2013 (H7N9) and A/Shanghai/1/2013 (H7N9) viruses. We thank the IMSUT serum bank for providing human sera. We thank R. Webster for providing monoclonal antibody to A/seal/Massachusetts/1/80 (H7N7). Polyclonal anti-influenza virus H7 HA, A/Netherlands/219/2003 (H7N7) (anti-serum, goat) NR-9226, was obtained through the National Institutes of Health (NIH) Biodefense and Emerging Infections Research Resources Repository, National Institute of Allergy and Infectious Diseases (NIAID), NIH. We thank S. Watson for editing the manuscript, T. Suzuki, K. Takahashi, S. Fujisaki and H. Xu for discussions, and Y. Sato, H. Sugawara, A. Sato, M. Ejima and T. Miura for technical assistance. We thank Toyama Chemical Co. for providing favipiravir, Daiichi Sankyo Co. for providing laninamivir, F. Hoffmann-La Roche for providing oseltamivir carboxylate, GlaxoSmithKline for providing zanamivir and Shionogi & Co. for providing peramivir. This work was supported by the Japan Initiative for Global Research Network on Infectious Diseases from the Ministry of Education, Culture, Sports, Science and Technology, Japan; by grants-in-aid from the Ministry of Health, Labour and Welfare, Japan; by ERATO (Japan Science and Technology Agency); by NIAID Public Health Service research grants AI099274 and AI058113 to J.C.P., and by an NIAID-funded Center for Research on Influenza Pathogenesis (CRIP, HHSN266200700010C) to Y.K.

Author information

Author notes

    • Tokiko Watanabe
    • , Maki Kiso
    • , Satoshi Fukuyama
    • , Noriko Nakajima
    •  & Masaki Imai

    These authors contributed equally to this work.

Affiliations

  1. ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 332-0012, Japan

    • Tokiko Watanabe
    • , Satoshi Fukuyama
    • , Dongming Zhao
    • , Yuriko Tomita
    • , Naomi Fujimoto
    • , Kazue Goto
    • , Eiryo Kawakami
    • , Izumi Ishikawa
    • , Shinji Watanabe
    •  & Yoshihiro Kawaoka
  2. Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan

    • Maki Kiso
    • , Shinya Yamada
    • , Seiya Yamayoshi
    • , Kiyoko Iwatsuki-Horimoto
    • , Takeshi Noda
    • , Hiroaki Katsura
    • , Mutsumi Ito
    • , Yuko Sakai-Tagawa
    • , Yukihiko Sugita
    • , Ryuta Uraki
    • , Reina Yamaji
    •  & Yoshihiro Kawaoka
  3. Department of Pathology, National Institute of Infectious Diseases, Shinjuku, Tokyo 162-8640, Japan

    • Noriko Nakajima
    •  & Hideki Hasegawa
  4. Influenza Virus Research Center, National Institute of Infectious Diseases, Musashimurayama, Tokyo 208-0011, Japan

    • Masaki Imai
    • , Emi Takashita
    • , Noriko Kishida
    • , Masayuki Shirakura
    • , Takato Odagiri
    •  & Masato Tashiro
  5. Department of Special Pathogens, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan

    • Shin Murakami
    •  & Yoshihiro Kawaoka
  6. Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan

    • Yoshihiro Sakoda
    • , Shintaro Shichinohe
    • , Masatoshi Okamatsu
    • , Tomokazu Tamura
    •  & Hiroshi Kida
  7. The Scripps Research Institute, 10550 North Torrey Pines Road, SP-3 La Jolla, California 92037, USA

    • Ryan McBride
    • , Robert P. de Vries
    •  & James C. Paulson
  8. Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison, Wisconsin 53711, USA

    • Masato Hatta
    • , Hirotaka Imai
    • , Amie J. Eisfeld
    • , Gongxun Zhong
    • , Shufang Fan
    • , Jihui Ping
    • , Eileen A. Maher
    • , Anthony Hanson
    • , Gabriele Neumann
    •  & Yoshihiro Kawaoka
  9. Laboratory of Veterinary Microbiology, Department of Veterinary Sciences, University of Miyazaki, Miyazaki 889-2192, Japan

    • Shinji Watanabe
  10. Influenza and Prion Disease Research Center, National Institute of Animal Health, Kannondai 3-1-5, Tsukuba, Ibaraki 305-0856, Japan

    • Yuko Uchida
    •  & Takehiko Saito
  11. Laboratory of Animal Hygiene, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, Japan

    • Makoto Ozawa
  12. Transboundary Animal Disease Center, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, Japan

    • Makoto Ozawa
  13. Research Center for Zoonosis Control, Hokkaido University, Sapporo 001-0020, Japan

    • Hiroshi Kida
  14. Laboratory of Bioresponses Regulation, Department of Biological Responses, Institute for Virus Research, Kyoto University, Kyoto 606-8507, Japan

    • Yoshihiro Kawaoka

Authors

  1. Search for Tokiko Watanabe in:

  2. Search for Maki Kiso in:

  3. Search for Satoshi Fukuyama in:

  4. Search for Noriko Nakajima in:

  5. Search for Masaki Imai in:

  6. Search for Shinya Yamada in:

  7. Search for Shin Murakami in:

  8. Search for Seiya Yamayoshi in:

  9. Search for Kiyoko Iwatsuki-Horimoto in:

  10. Search for Yoshihiro Sakoda in:

  11. Search for Emi Takashita in:

  12. Search for Ryan McBride in:

  13. Search for Takeshi Noda in:

  14. Search for Masato Hatta in:

  15. Search for Hirotaka Imai in:

  16. Search for Dongming Zhao in:

  17. Search for Noriko Kishida in:

  18. Search for Masayuki Shirakura in:

  19. Search for Robert P. de Vries in:

  20. Search for Shintaro Shichinohe in:

  21. Search for Masatoshi Okamatsu in:

  22. Search for Tomokazu Tamura in:

  23. Search for Yuriko Tomita in:

  24. Search for Naomi Fujimoto in:

  25. Search for Kazue Goto in:

  26. Search for Hiroaki Katsura in:

  27. Search for Eiryo Kawakami in:

  28. Search for Izumi Ishikawa in:

  29. Search for Shinji Watanabe in:

  30. Search for Mutsumi Ito in:

  31. Search for Yuko Sakai-Tagawa in:

  32. Search for Yukihiko Sugita in:

  33. Search for Ryuta Uraki in:

  34. Search for Reina Yamaji in:

  35. Search for Amie J. Eisfeld in:

  36. Search for Gongxun Zhong in:

  37. Search for Shufang Fan in:

  38. Search for Jihui Ping in:

  39. Search for Eileen A. Maher in:

  40. Search for Anthony Hanson in:

  41. Search for Yuko Uchida in:

  42. Search for Takehiko Saito in:

  43. Search for Makoto Ozawa in:

  44. Search for Gabriele Neumann in:

  45. Search for Hiroshi Kida in:

  46. Search for Takato Odagiri in:

  47. Search for James C. Paulson in:

  48. Search for Hideki Hasegawa in:

  49. Search for Masato Tashiro in:

  50. Search for Yoshihiro Kawaoka in:

Contributions

T.W., M.K., S. Fukuyama, M. Imai, S. Yamada, S.M., S. Yamayoshi, K.I.-H., Y. Sakoda, E.T., M.H., S.W., E.A.M., G.N., H.K., T.O., J.C.P., M.T. and Y.K. designed the study; T.W., M.K., S. Fukuyama, N.N., M. Imai, S. Yamada, S.M., S. Yamayoshi, K.I.-H., Y. Sakoda, E.T., R.M., T.N., M.H., H.I., D.Z., N.K., M.S., R.P.d.V., S.S., M. Okamatsu., T.T., Y.T., N.F., K.G., H.K., I.I., M. Ito, Y.S.-T., Y. Sugita, R.U., R.Y., A.J.E., G.Z., S. Fang, J.P., A.H., Y.U., T.S. and H.H. performed the experiments; T.W., M.K., S. Fang, N.N., M. Imai, S. Yamayoshi, S.M., S. Yamada, K.I.-H., Y. Sakoda, E.T., R.M., T.N., M.H., H.I., D.Z., R.P.V., S.S., T.T., Y.T., H.K., E.K. and H. H. analysed the data; T.W., S. Fukuyama, N.N., E.T., R.M., M.H., R.P.d.V., M. Ozawa, G.N., T.O., J.C.P., H.H., M.T. and Y.K. wrote the manuscript; Y.K. oversaw the project. T.W., M.K., S. Fukuyama, N.N. and M. Imai contributed equally to this work.

Competing interests

Y.K. has received speaker’s honoraria from Chugai Pharmaceuticals, Novartis, Daiichi-Sankyo Pharmaceutical, Toyama Chemical, Wyeth, GlaxoSmithKline and Astellas; grant support from Chugai Pharmaceuticals, Daiichi Sankyo Pharmaceutical, Toyama Chemical and Otsuka Pharmaceutical Co.; is a consultant for Crucell; and is a founder of FluGen. G.N. is a founder of FluGen.

Corresponding author

Correspondence to Yoshihiro Kawaoka.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Results, Supplementary Figures S1-S11, Supplementary Tables S1-S18 and a Supplementary Reference.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature12392

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.