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Identification of nucleolin as a cellular receptor for human respiratory syncytial virus

Nature Medicine volume 17, pages 1132–1135 (2011)Cite this article

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Abstract

Human respiratory syncytial virus (RSV) causes a large burden of disease worldwide1. There is no effective vaccine or therapy, and the use of passive immunoprophylaxis with RSV-specific antibodies is limited to high-risk patients2,3,4,5. The cellular receptor (or receptors) required for viral entry and replication has yet to be described; its identification will improve understanding of the pathogenesis of infection and provide a target for the development of novel antiviral interventions. Here we show that RSV interacts with host-cell nucleolin via the viral fusion envelope glycoprotein and binds specifically to nucleolin at the apical cell surface in vitro. We observed decreased RSV infection in vitro in neutralization experiments using nucleolin-specific antibodies before viral inoculation, in competition experiments in which virus was incubated with soluble nucleolin before inoculation of cells, and upon RNA interference (RNAi) to silence cellular nucleolin expression. Transfection of nonpermissive Spodoptera frugiperda Sf9 insect cells with human nucleolin conferred susceptibility to RSV infection. RNAi-mediated knockdown of lung nucleolin was associated with a significant reduction in RSV infection in mice (P = 0.0004), confirming that nucleolin is a functional RSV receptor in vivo.

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Figure 1: RSV VOPBA and coimmunoprecipitation of RSV fusion protein with nucleolin.
Figure 2: RSV and nucleolin colocalized on cell surfaces, and inhibition interventions decreased infection.
Figure 3: Sf9 cells are made permissive to RSV infection by heterologous expression of human nucleolin.
Figure 4: Silencing nucleolin reduces RSV infection in mice.

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Acknowledgements

The authors thank R.A. Weiss, B. McManus and F. Jean for helpful comments, and L. Morikawa and Q. Xu for help with immunohistochemical staining of mouse lung. F.T. is the recipient of a Canadian Institutes of Health Research (CIHR) Institute of Genetics Grant for Short-term Visits, and D.M. is the recipient of CIHR IMPACT and Heart and Stroke Foundation of British Columbia Post-doctoral Fellowship awards.

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Author notes

  1. Farnoosh Tayyari and David Marchant: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, The James Hogg Research Centre, Providence Heart and Lung Institute at St. Paul's Hospital, University of British Columbia, Vancouver, British Columbia, Canada

    Farnoosh Tayyari & David Marchant

  2. Department of Paediatrics, The Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada

    Theo J Moraes & Wenming Duan

  3. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada

    Theo J Moraes, Peter Mastrangelo & Richard G Hegele

  4. Department of Paediatric Laboratory Medicine, The Hospital for Sick Children, Toronto, Ontario, Canada

    Richard G Hegele

Authors
  1. Farnoosh Tayyari
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  2. David Marchant
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  3. Theo J Moraes
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Contributions

F.T. conducted VOPBAs, mass spectrometry analysis, neutralization and competition experiments in vitro. D.M. conducted immunoprecipitation, cell culture and virus experiments with mammalian and Sf9 cells. T.J.M. performed mouse dissections and lung fixation. W.D. generated virus for the mouse experiments and did quantitative plaque assays. P.M. assisted with cell culture and mouse experiments, analyzed the data, prepared figures and graphs, and coordinated manuscript writing. R.G.H. did mouse lung histological examination and supervised the project.

Corresponding author

Correspondence to Richard G Hegele.

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The authors declare no competing financial interests.

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Supplementary Figures 1–13, Supplementary Table 1 and Supplementary Methods (PDF 1429 kb)

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Tayyari, F., Marchant, D., Moraes, T. et al. Identification of nucleolin as a cellular receptor for human respiratory syncytial virus. Nat Med 17, 1132–1135 (2011). https://doi.org/10.1038/nm.2444

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