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The GD1a glycan is a cellular receptor for adenoviruses causing epidemic keratoconjunctivitis

Nature Medicine volume 17, pages 105–109 (2011)Cite this article

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Abstract

Adenovirus type 37 (Ad37) is a leading cause of epidemic keratoconjunctivitis (EKC)1,2, a severe and highly contagious ocular disease. Whereas most other adenoviruses infect cells by engaging CD46 or the coxsackie and adenovirus receptor (CAR), Ad37 binds previously unknown sialic acid–containing cell surface molecules3,4. By glycan array screening, we show here that the receptor-recognizing knob domain of the Ad37 fiber protein specifically binds a branched hexasaccharide that is present in the GD1a ganglioside and that features two terminal sialic acids. Soluble GD1a glycan and GD1a-binding antibodies efficiently prevented Ad37 virions from binding and infecting corneal cells. Unexpectedly, the receptor is constituted by one or more glycoproteins containing the GD1a glycan motif rather than the ganglioside itself, as shown by binding, infection and flow cytometry experiments. Molecular modeling, nuclear magnetic resonance and X-ray crystallography reveal that the two terminal sialic acids dock into two of three previously established sialic acid–binding sites in the trimeric Ad37 knob. Surface plasmon resonance analysis shows that the knob–GD1a glycan interaction has high affinity. Our findings therefore form a basis for the design and development of sialic acid–containing antiviral drugs for topical treatment of EKC.

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Figure 1: Glycan array of Ad37 knob interactions with 260 different glycans.
Figure 2: Ad37 binding to the corneal epithelial cell surface depends on fiber knob binding to molecules containing the GD1a glycan.
Figure 3: X-ray crystallography and functional analysis of the Ad37 knob-GD1a glycan complex.
Figure 4: Surface plasmon resonance analysis of GD1a glycan binding to immobilized Ad37 knob protein.

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Acknowledgements

We highly appreciate the support from F. Lindh and S. Spjut regarding the purification of GD1a glycan; F. Lindh (Isosep) provided GD1a gangliosides. We also thank A. Carlsson (Medigelium) for providing GD1a-containing liposomes; D. Guilligay (EMBL) for providing the Ad37 knob gene cloned into pPROEX Htb plasmid (Life Technologies)16; R.L. Schnaar (The Johns Hopkins University School of Medicine) for providing P4 compounds; and K. Lindman, M. Hägg and F. Jamshidi for technical support. HCE cells were provided by K. Araki-Sasaki (Kinki Central Hospital). GM3 and GD2 glycans were provided by the Consortium for Functional Glycomics. We also highly appreciate the resources provided by the Consortium for Functional Glycomics (funded by National Institute of General Medical Sciences grant no. GM62116) Core D and H, as well as related technical support from O. Blixt and N. Reza. We are grateful to the Berlin Electron Storage Ring Society for Synchrotron Radiation (BESSY) for beamtime and beamline support. This project was supported by the Swedish Research Council (grants no. 2007-3402 (N.A.); 2009-3859 (N.A.) and 11612 (J.Å. via M.E. Breimer)), the Swedish Foundation for Strategic Research (grant no. F06-0011 to N.A.), the Swedish Society of Medicine (grant no. 97031 to N.A.), the Estonian Science Foundation (grant 8300 to A.L.), the Collaborative Research Center SFB-685 (T.S.), a student fellowship from the University of Tübingen (J.B.) and The Wellcome Trust (S.R. and H.J.W.).

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Authors and Affiliations

  1. Department of Clinical Microbiology, Division of Virology, Umeå University, Umeå, Sweden

    Emma C Nilsson, Rickard J Storm, Susanne M C Johansson, Therese L Eriksson, Lars Frängsmyr & Niklas Arnberg

  2. Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany

    Johannes Bauer & Thilo Stehle

  3. Department of Chemistry, Tallinn University of Technology, Tallinn, Estonia

    Aivar Lookene

  4. Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, University of Göteborg, Göteborg, Sweden

    Jonas Ångström

  5. Department of Chemistry, Computational Life Science Cluster (CLiC), Umeå University, Umeå, Sweden

    Mattias Hedenström

  6. Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, Glasgow Biomedical Research Centre, University of Glasgow, Glasgow, UK

    Simon Rinaldi & Hugh J Willison

  7. Department of Clinical Sciences, Ophthalmology, Umeå University, Umeå, Sweden

    Fatima Pedrosa Domellöf

  8. Department of Integrative Medical Biology, Anatomy, Umeå University, Umeå, Sweden

    Fatima Pedrosa Domellöf

  9. Department of Pediatrics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Thilo Stehle

  10. Laboratory for Molecular Infection Medicine in Sweden (MIMS), Umeå University, Umeå, Sweden

    Niklas Arnberg

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  1. Emma C Nilsson
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Contributions

E.C.N. and R.J.S. contributed equally to design and conduction of binding, infection and flow cytometry experiments; S.M.C.J. produced GD1a glycan and performed NMR studies with M.H.; J.B. and T.S. carried out X-ray crystallography studies; S.R. and H.J.W. performed combinatorial glycolipid glycoarray; J.Å. conducted molecular modeling; F.P.D. carried out immunohistochemistry analysis; and A.L. performed SPR experiments. L.F. and T.L.E. conducted two-dimensional gel electrophoresis and blotting experiments, and L.F. did statistical calculations. T.S., M.H., F.P.D., A.L., J.Å., H.J.W., L.F., J.B. and N.A. discussed and wrote the manuscript, and T.S. and N.A. supervised the project.

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Correspondence to Niklas Arnberg.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6 and Supplementary Tables 2–5 (PDF 4855 kb)

Supplementary Table 1

Summary of glycan array data (XLS 119 kb)

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Nilsson, E., Storm, R., Bauer, J. et al. The GD1a glycan is a cellular receptor for adenoviruses causing epidemic keratoconjunctivitis. Nat Med 17, 105–109 (2011). https://doi.org/10.1038/nm.2267

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