• A Corrigendum to this article was published on 26 September 2012

Abstract

Investigation of the human antibody response to influenza virus infection has been largely limited to serology, with relatively little analysis at the molecular level. The 1918 H1N1 influenza virus pandemic was the most severe of the modern era1. Recent work has recovered the gene sequences of this unusual strain2, so that the 1918 pandemic virus could be reconstituted to display its unique virulence phenotypes3,4. However, little is known about adaptive immunity to this virus. We took advantage of the 1918 virus sequencing and the resultant production of recombinant 1918 haemagglutinin (HA) protein antigen to characterize at the clonal level neutralizing antibodies induced by natural exposure of survivors to the 1918 pandemic virus. Here we show that of the 32 individuals tested that were born in or before 1915, each showed seroreactivity with the 1918 virus, nearly 90 years after the pandemic. Seven of the eight donor samples tested had circulating B cells that secreted antibodies that bound the 1918 HA. We isolated B cells from subjects and generated five monoclonal antibodies that showed potent neutralizing activity against 1918 virus from three separate donors. These antibodies also cross-reacted with the genetically similar HA of a 1930 swine H1N1 influenza strain, but did not cross-react with HAs of more contemporary human influenza viruses. The antibody genes had an unusually high degree of somatic mutation. The antibodies bound to the 1918 HA protein with high affinity, had exceptional virus-neutralizing potency and protected mice from lethal infection. Isolation of viruses that escaped inhibition suggested that the antibodies recognize classical antigenic sites on the HA surface. Thus, these studies demonstrate that survivors of the 1918 influenza pandemic possess highly functional, virus-neutralizing antibodies to this uniquely virulent virus, and that humans can sustain circulating B memory cells to viruses for many decades after exposure—well into the tenth decade of life.

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Data deposits

Antibody nucleotide sequences have been deposited in GenBank under accession numbers EU169674 to EU169679, and EU825947 to EU825950.

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Acknowledgements

We thank L. Schoenherr, K. Sharma, L. Adams (supported by the University of Medicine and Dentistry of New Jersey (UMDNJ) Institute for the Elimination of Health Disparities), C. Dokes, C. Gaines, B. Butter and S. Rivera for assistance with subjects, and S. Yoder and B. Briney for sample preparation. This work was supported by grants from the National Institutes of Health to J.E.C. (U54 AI057157, Southeast Regional Center of Excellence for Emerging Infections and Biodefense, and U19 AI057229), and C.F.B. (U54 AI57158, Northeast Biodefense Center, U19 AI62623, Center for Investigating Viral Immunity and Antagonism, and P01 AI058113). I.A.W. and J.S. were supported in part by the National Institutes of Health (CA55896 and AI42266). P.V.A. was supported by a fellowship awarded by the Northeast Biodefense Center (AI057158). We thank P. Palese and A. Garcia-Sastre for advice and for providing H1N1 viruses, and M. Posner and L. Cavacini for the HMMA2.5 cell line.

Author Contributions X.Y., P.A.M., M.D.H. and F.S.H. made and cloned the monoclonal antibodies, sequenced antibody genes, and performed immunofluorescence experiments. T.T. characterized the interaction of the antibodies with viruses and VLPs and selected for and characterized the escape mutants. C.J.K. performed biosensor studies. T.M.T., C.P. and L.A.P. designed and performed in vivo studies. O.M. sequenced the HA genes of the H1N1 viruses used in this study and performed ELISA assays with these viruses. P.V.A. assisted with HAI and neutralization assays and with cloning of recombinant HA molecules. J.S. and I.A.W. provided recombinant HA. E.L.A. led the clinical recruitment and, together with C.F.B. and J.E.C., conceived of the experimental plan. C.F.B. and J.E.C. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Author information

Author notes

    • Xiaocong Yu
    •  & Tshidi Tsibane

    These authors contributed equally to this work.

Affiliations

  1. Departments of Pediatrics and of Microbiology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee 37232, USA

    • Xiaocong Yu
    • , Patricia A. McGraw
    • , Frances S. House
    • , Christopher J. Keefer
    • , Mark D. Hicar
    •  & James E. Crowe Jr
  2. Department of Microbiology, Mount Sinai School of Medicine, New York 10029, USA

    • Tshidi Tsibane
    • , Claudia Pappas
    • , Osvaldo Martinez
    • , Patricia V. Aguilar
    •  & Christopher F. Basler
  3. Influenza Division, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA

    • Terrence M. Tumpey
    • , Claudia Pappas
    • , Lucy A. Perrone
    •  & James Stevens
  4. Department of Molecular Biology and Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California 92037, USA

    • James Stevens
    •  & Ian A. Wilson
  5. Department of Physical Medicine and Rehabilitation, University of Medicine & Dentistry of New Jersey, Newark, New Jersey 07103, USA

    • Eric L. Altschuler

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Correspondence to Eric L. Altschuler or Christopher F. Basler or James E. Crowe Jr.

Supplementary information

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  1. 1.

    Supplementary Information

    The file contains Supplementary Tables 1-2 and Supplementary Figures and Legends 1-3.  The Supplementary Tables illustrate 1) Serologic data from volunteers of varying ages to 1918 and Sw/30 antigens and viruses, 2) Neutralization or HAI specific activity (µg/mL) of mAbs against representative H1N1 viruses.  The Supplementary Figures illustrate, 1) ELISA binding data for mAbs 1F1, 2D1, and 4D20 to representative 20th century H1N1 viruses, 2) binding of human mAbs to 1918 HA protein in HA transfected cells, and 3) the location of escape mutations in the HA for variant viruses selected with mAbs. This file was replaced on 27th September 2012 - see Corrigendum nature11235 linked to this paper for details.

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https://doi.org/10.1038/nature07231

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