Preexisting human antibodies neutralize recently emerged H7N9 influenza strains

Carole Dunand

Paul Leon

Kaval Kaur

Gene Tan

Yunping Huang+10 authors

Supplemental data:

Click here to view.^{(229K, pdf)}

^{Department of Medicine, Section of Rheumatology, Gwen Knapp Center for Lupus and Immunology Research, The University of Chicago, Chicago, Illinois, USA.}

^{Department of Microbiology and}

^{Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA.}

^{Committee on Immunology, The University of Chicago, Chicago, Illinois, USA.}

^{Emory Vaccine Center, Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA.}

^{Department of Medicine and}

^{Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA.}

Address correspondence to: Patrick C. Wilson, The University of Chicago, BSLC/Jules F. Knapp Building, 924 East 57th street, R414, Chicago, Illinois 60637, USA. Phone: 773.702.9009; E-mail: ude.ogacihcu@pnosliw. Or to: Florian Krammer, Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA. Phone: 212.241.7318; E-mail: ude.mssm@remmark.nairolf.

Authorship note: Patrick C. Wilson and Florian Krammer contributed equally to this work.

Received 2014 Jul 21; Accepted 2015 Jan 6.

Abstract

The emergence and seasonal persistence of pathogenic H7N9 influenza viruses in China have raised concerns about the pandemic potential of this strain, which, if realized, would have a substantial effect on global health and economies. H7N9 viruses are able to bind to human sialic acid receptors and are also able to develop resistance to neuraminidase inhibitors without a loss in fitness. It is not clear whether prior exposure to circulating human influenza viruses or influenza vaccination confers immunity to H7N9 strains. Here, we demonstrate that 3 of 83 H3 HA-reactive monoclonal antibodies generated by individuals that had previously undergone influenza A virus vaccination were able to neutralize H7N9 viruses and protect mice against homologous challenge. The H7N9-neutralizing antibodies bound to the HA stalk domain but exhibited a difference in their breadth of reactivity to different H7 influenza subtypes. Mapping viral escape mutations suggested that these antibodies bind at least two different epitopes on the stalk region. Together, these results indicate that these broadly neutralizing antibodies may contribute to the development of therapies against H7N9 strains and may also be effective against pathogenic H7 strains that emerge in the future.

Abstract

Acknowledgments

This work was supported in part by NIH grants 1U19AI08724 (to P.C. Wilson), HHSN272201400005C (to P.C. Wilson), 5U54AI057158 (to P.C. Wilson and R. Ahmed), 5U19AI057266 (to P.C. Wilson, R. Ahmed, and J. Wrammert), 1U19AI090023 (to P.C. Wilson, R. Ahmed, and J. Wrammert), 1P01AI097092 (to P.C. Wilson, R. Ahmed, P. Palese, and A. García-Sastre), and 1U19 {"type":"entrez-nucleotide","attrs":{"text":"AI109946","term_id":"3478270","term_text":"AI109946"}}AI109946 (to P.C. Wilson and P. Palese) and by funds provided by the Gwen Knapp Center for Lupus and Immunology Research. This work was also supported in part by CRIP (Center for Research on Influenza Pathogenesis), an NIAID-funded Center of Excellence for Influenza Research and Surveillance (contract HHSN272201400008C) (P. Palese, A. García-Sastre, F. Krammer). Florian Krammer was supported by an Erwin Schrödinger fellowship (J3232) from the Austrian Science Fund (FWF). Kaval Kaur was supported by a National Science Scholarship (PhD) from the Agency of Science, Technology and Research (A*STAR), Singapore. We thank Jori Reigle, Samuel Lim, Marissa Kumabe, Elizabeth Yan, and Lie Li for collecting blood samples; Meghan Sullivan, Noel Pauli, and Andrew Kinloch for helpful comments on the manuscript; and Ariana Hirsh for technical assistance.

Acknowledgments

Footnotes

Conflict of interest: The authors have declared that no conflict of interest exists.

Reference information:J Clin Invest. 2015;125(3):1255–1268. doi:10.1172/JCI74374.

Footnotes

References

1. Shaw ML Palese P. Orthomyxoviruses. In: Knipe DM, Howley PM, eds. Fields Virology. 6th ed. Philadelphia, Pennsylvania, USA: Lippincott Williams and Wilkins; 2013:1151–1185. [PubMed]
2. Kaur K, Sullivan M, Wilson PCTargeting B cell responses in universal influenza vaccine design. Trends Immunol. 2011;32(11):524–531. doi: 10.1016/j.it.2011.08.007.] [[Google Scholar]
3. Air GMSequence relationships among the hemagglutinin genes of 12 subtypes of influenza A virus. Proc Natl Acad Sci U S A. 1981;78(12):7639–7643. doi: 10.1073/pnas.78.12.7639.] [[Google Scholar]
4. Medina RA, Garcia-Sastre AInfluenza A viruses: new research developments. Nat Rev Microbiol. 2011;9(8):590–603. doi: 10.1038/nrmicro2613.] [[PubMed][Google Scholar]
5. Hirst M, et al Novel avian influenza H7N3 strain outbreak, British Columbia. Emerg Infect Dis. 2004;10(12):2192–2195. doi: 10.3201/eid1012.040743.] [[Google Scholar]
6. Fouchier RA, et al Avian influenza A virus (H7N7) associated with human conjunctivitis and a fatal case of acute respiratory distress syndrome. Proc Natl Acad Sci U S A. 2004;101(5):1356–1361. doi: 10.1073/pnas.0308352100.] [[Google Scholar]
7. Gao R, et al Human infection with a novel avian-origin influenza A (H7N9) virus. N Engl J Med. 2013;368(20):1888–1897. doi: 10.1056/NEJMoa1304459.] [[PubMed][Google Scholar]
8. Belser JA, et al Pathogenesis transmission of avian influenza A (H7N9) virus in ferrets mice. Nature. 2013;501(7468):556–559. doi: 10.1038/nature12391.] [[PubMed][Google Scholar]
9. Watanabe T, et al Characterization of H7N9 influenza A viruses isolated from humans. Nature. 2013;501(7468):551–555. doi: 10.1038/nature12392.] [[Google Scholar]
10. Morens DM, Taubenberger JK, Fauci ASH7N9 avian influenza A virus and the perpetual challenge of potential human pandemicity. MBio. 2013;4(4):1–14.[Google Scholar]
11. Hu Y, et al Association between adverse clinical outcome in human disease caused by novel influenza A H7N9 virus and sustained viral shedding and emergence of antiviral resistance. Lancet. 2013;381(9885):2273–2279. doi: 10.1016/S0140-6736(13)61125-3.] [[PubMed][Google Scholar]
12. Zhang Q, et al H7N9 influenza viruses are transmissible in ferrets by respiratory droplet. Science. 2013;341(6144):410–414. doi: 10.1126/science.1240532.] [[PubMed][Google Scholar]
13. Xu R, et al Preferential recognition of avian-like receptors in human influenza A H7N9 viruses. Science. 2013;342(6163):1230–1235. doi: 10.1126/science.1243761.] [[Google Scholar]
14. Ramos I, et al H7N9 influenza viruses interact preferentially with α2,3-linked sialic acids and bind weakly to α2,6-linked sialic acids. J Gen Virol. 2013;94(pt 11):2417–2423.[Google Scholar]
15. Hai R, et al Influenza A(H7N9) virus gains neuraminidase inhibitor resistance without loss of in vivo virulence or transmissibility. Nat Commun. 2013;4:2854.[Google Scholar]
16. Wrammert J, et al Broadly cross-reactive antibodies dominate the human B cell response against 2009 pandemic H1N1 influenza virus infection. J Exp Med. 2011;208(1):181–193. doi: 10.1084/jem.20101352.] [[Google Scholar]
17. Wrammert J, et al Rapid cloning of high-affinity human monoclonal antibodies against influenza virus. Nature. 2008;453(7195):667–671. doi: 10.1038/nature06890.] [[Google Scholar]
18. Krammer F, et al An H7N1 influenza virus vaccine induces broadly reactive antibody responses against H7N9 in humans. Clin Vaccine Immunol. 2014;21(8):1153–1163. doi: 10.1128/CVI.00272-14.] [[Google Scholar]
19. Dreyfus C, et al Highly conserved protective epitopes on influenza B viruses. Science. 2012;337(6100):1343–1348. doi: 10.1126/science.1222908.] [[Google Scholar]
20. Kapczynski DR, et al Characterization of the 2012 highly pathogenic avian influenza H7N3 virus isolated from poultry in an outbreak in Mexico: pathobiology and vaccine protection. J Virol. 2013;87(16):9086–9096. doi: 10.1128/JVI.00666-13.] [[Google Scholar]
21. Brandenburg B, et al Mechanisms of hemagglutinin targeted influenza virus neutralization. PLoS One. 2013;8(12):e80034. doi: 10.1371/journal.pone.0080034.] [[Google Scholar]
22. Tan GS, et al Characterization of a broadly neutralizing monoclonal antibody that targets the fusion domain of group 2 influenza a virus hemagglutinin. J Virol. 2014;88(23):13580–13592. doi: 10.1128/JVI.02289-14.] [[Google Scholar]
23. Ekiert DC, et al A highly conserved neutralizing epitope on group 2 influenza A viruses. Science. 2011;333(6044):843–850. doi: 10.1126/science.1204839.] [[Google Scholar]
24. Hensley SE, et al Hemagglutinin receptor binding avidity drives influenza A virus antigenic drift. Science. 2009;326(5953):734–736. doi: 10.1126/science.1178258.] [[Google Scholar]
25. Corti D, et al Heterosubtypic neutralizing antibodies are produced by individuals immunized with a seasonal influenza vaccine. J Clin Invest. 2010;120(5):1663–1673. doi: 10.1172/JCI41902.] [[Google Scholar]
26. Li GM, et al Pandemic H1N1 influenza vaccine induces a recall response in humans that favors broadly cross-reactive memory B cells. Proc Natl Acad Sci U S A. 2012;109(23):9047–9052. doi: 10.1073/pnas.1118979109.] [[Google Scholar]
27. Thomson CA, et al Pandemic H1N1 influenza infection and vaccination in humans induces cross-protective antibodies that target the hemagglutinin stem. Front Immunol. 2012;3:87.[Google Scholar]
28. Wilson PC, Andrews SFTools to therapeutically harness the human antibody response. Nat Rev Immunol. 2012;12(10):709–719. doi: 10.1038/nri3285.] [[PubMed][Google Scholar]
29. Ekiert DC, et al Cross-neutralization of influenza A viruses mediated by a single antibody loop. Nature. 2012;489(7417):526–532. doi: 10.1038/nature11414.] [[Google Scholar]
30. Margine I, et al H3N2 influenza virus infection induces broadly reactive hemagglutinin stalk antibodies in humans and mice. J Virol. 2013;87(8):4728–4737. doi: 10.1128/JVI.03509-12.] [[Google Scholar]
31. Margine I, et al Hemagglutinin stalk-based universal vaccine constructs protect against group 2 influenza A viruses. J Virol. 2013;87(19):10435–10446. doi: 10.1128/JVI.01715-13.] [[Google Scholar]
32. Krammer F, et al H3 stalk-based chimeric hemagglutinin influenza virus constructs protect mice from H7N9 challenge. J Virol. 2014;88(4):2340–2343. doi: 10.1128/JVI.03183-13.] [[Google Scholar]
33. Ekiert DC, et al Antibody recognition of a highly conserved influenza virus epitope. Science. 2009;324(5924):246–251. doi: 10.1126/science.1171491.] [[Google Scholar]
34. Friesen RH, et al A common solution to group 2 influenza virus neutralization. Proc Natl Acad Sci U S A. 2014;111(1):445–450. doi: 10.1073/pnas.1319058110.] [[Google Scholar]
35. O’Donnell CD, et al Antibody pressure by a human monoclonal antibody targeting the 2009 pandemic H1N1 virus hemagglutinin drives the emergence of a virus with increased virulence in mice. MBio. 201233e00120-12 [Google Scholar]
36. Wang TT, et al Broadly protective monoclonal antibodies against H3 influenza viruses following sequential immunization with different hemagglutinins. PLoS Pathog. 2010;6(2):e1000796. doi: 10.1371/journal.ppat.1000796.] [[Google Scholar]
37. Corti D, et al A neutralizing antibody selected from plasma cells that binds to group 1 and group 2 influenza A hemagglutinins. Science. 2011;333(6044):850–856. doi: 10.1126/science.1205669.] [[PubMed][Google Scholar]
38. Throsby M, et al Heterosubtypic neutralizing monoclonal antibodies cross-protective against H5N1 and H1N1 recovered from human IgM^{memory B cells.}PLoS One. 2008;3(12):e3942. doi: 10.1371/journal.pone.0003942.] [[Google Scholar]
39. Sui J, et al Structural and functional bases for broad-spectrum neutralization of avian and human influenza A viruses. Nat Struct Mol Biol. 2009;16(3):265–273. doi: 10.1038/nsmb.1566.] [[Google Scholar]
40. Krammer F, et al Divergent H7 immunogens offer protection from H7N9 virus challenge. J Virol. 2014;88(8):3976–3985. doi: 10.1128/JVI.03095-13.] [[Google Scholar]
41. Margine I, Palese P, Krammer FExpression of functional recombinant hemagglutinin and neuraminidase proteins from the novel H7N9 influenza virus using the baculovirus expression system. J Vis Exp. 2013;(81):e51112.[Google Scholar]
42. Crotty S, Aubert RD, Glidewell J, Ahmed RTracking human antigen-specific memory B cells: a sensitive and generalized ELISPOT system. J Immunol Methods. 2004;286(1-2):111–122. doi: 10.1016/j.jim.2003.12.015.] [[PubMed][Google Scholar]
43. Quinlivan M, Zamarin D, Garcia-Sastre A, Cullinane A, Chambers T, Palese PAttenuation of equine influenza viruses through truncations of the NS1 protein. J Virol. 2005;79(13):8431–8439. doi: 10.1128/JVI.79.13.8431-8439.2005.] [[Google Scholar]
44. Fodor E, Devenish L, Engelhardt OG, Palese P, Brownlee GG, Garcia-Sastre ARescue of influenza A virus from recombinant DNA. J Virol. 1999;73(11):9679–9682.[Google Scholar]
45. Hai R, et al Influenza viruses expressing chimeric hemagglutinins: globular head and stalk domains derived from different subtypes. J Virol. 2012;86(10):5774–5781. doi: 10.1128/JVI.00137-12.] [[Google Scholar]
46. Smith K, et al Rapid generation of fully human monoclonal antibodies specific to a vaccinating antigen. Nat Protoc. 2009;4(3):372–384. doi: 10.1038/nprot.2009.3.] [[Google Scholar]