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Rapid cloning of high-affinity human monoclonal antibodies against influenza virus

Nature volume 453, pages 667–671 (2008)Cite this article

Abstract

Pre-existing neutralizing antibody provides the first line of defence against pathogens in general. For influenza virus, annual vaccinations are given to maintain protective levels of antibody against the currently circulating strains. Here we report that after booster vaccination there was a rapid and robust influenza-specific IgG+ antibody-secreting plasma cell (ASC) response that peaked at approximately day 7 and accounted for up to 6% of peripheral blood B cells. These ASCs could be distinguished from influenza-specific IgG+ memory B cells that peaked 14–21 days after vaccination and averaged 1% of all B cells. Importantly, as much as 80% of ASCs purified at the peak of the response were influenza specific. This ASC response was characterized by a highly restricted B-cell receptor (BCR) repertoire that in some donors was dominated by only a few B-cell clones. This pauci-clonal response, however, showed extensive intraclonal diversification from accumulated somatic mutations. We used the immunoglobulin variable regions isolated from sorted single ASCs to produce over 50 human monoclonal antibodies (mAbs) that bound to the three influenza vaccine strains with high affinity. This strategy demonstrates that we can generate multiple high-affinity mAbs from humans within a month after vaccination. The panel of influenza-virus-specific human mAbs allowed us to address the issue of original antigenic sin (OAS): the phenomenon where the induced antibody shows higher affinity to a previously encountered influenza virus strain compared with the virus strain present in the vaccine1. However, we found that most of the influenza-virus-specific mAbs showed the highest affinity for the current vaccine strain. Thus, OAS does not seem to be a common occurrence in normal, healthy adults receiving influenza vaccination.

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Figure 1: Analysis of the B-cell response induced by influenza vaccination.
Figure 2: The ASC response after influenza vaccination is pauci-clonal and highly diversified by somatic hypermutation.
Figure 3: High-affinity mAbs generated from single influenza-specific ASCs.
Figure 4: Specificity for the newly introduced influenza-B strain in the vaccine suggests a minimal impact of OAS.

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Acknowledgements

We thank A. Popkowski, H. Wu, L. Abraham and B. Begley for technical assistance, and R. Casellas and J. Knight for reading the manuscript. This work was funded in parts by National Institutes of Health (NIH) grant numbers HHSN266200500026C (P.C.W.), P20 RR018758 (P.C.W.), NIH/National Institute of Allergy and Infectious Diseases (NIAID) U19-AI057266-04 (R.A.), NIH/NIAID HHSN266200700006C Center of Excellence for Influenza Research and Surveillance (R.A.) and NIH/NIAID N01-AI-50025-02 (R.A. and C.L.). J.W. was supported by a postdoctoral fellowship from The Swedish Research Council.

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Author notes

  1. Jens Wrammert and Kenneth Smith: These authors contributed equally to this work.

Authors and Affiliations

  1. Emory Vaccine Center and Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA,

    Jens Wrammert, Joe Miller, William A. Langley & Rafi Ahmed

  2. Emory Transplant Center and Department of Surgery, Emory University School of Medicine, Atlanta, Georgia 30322, USA,

    Kenneth Kokko & Christian Larsen

  3. Immunobiology and Cancer Research Program, The Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA,

    Kenneth Smith, Nai-Ying Zheng, Israel Mays, Lori Garman, Christina Helms & Patrick C. Wilson

  4. Arthritis and Immunology Research Program, The Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA,

    Judith James

  5. Molecular Immunogenetics Research Program, The Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA,

    J. Donald Capra

  6. Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA,

    Judith James & Patrick C. Wilson

  7. Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA,

    Judith James

  8. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA,

    Gillian M. Air

  9. Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA,

    J. Donald Capra & Patrick C. Wilson

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Corresponding author

Correspondence to Patrick C. Wilson.

Supplementary information

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The file contains Supplementary Figures 1-4 with Legends, Supplementary Tables 1-2 and Supplementary Methods (PDF 3828 kb)

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Wrammert, J., Smith, K., Miller, J. et al. Rapid cloning of high-affinity human monoclonal antibodies against influenza virus. Nature 453, 667–671 (2008). https://doi.org/10.1038/nature06890

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