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Receptor-binding specificity of a bovine influenza A virus

Nature volume 640, pages E21–E27 (2025)Cite this article

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Matters Arising to this article was published on 16 April 2025

The Original Article was published on 08 July 2024

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arising from: A. J. Eisfeld et al. Nature https://doi.org/10.1038/s41586-024-07766-6 (2024).

The receptor-binding preference of haemagglutinin of influenza A viruses (IAVs) is a major determinant of host range1,2, and avian viruses preferentially bind α2,3-linked sialic acids, whereas human IAVs recognize α2,6-linked sialic acids. Eisfeld et al.3 examined the pathogenicity and transmissibility of a highly pathogenic avian influenza (HPAI) bovine H5N1 virus, and found that it has a dual binding specificity for human and avian receptors, as measured in a solid-phase glycan-binding assay, raising concerns about its pandemic potential. Here we examined the receptor specificity of a bovine HPAI H5N1 virus (A/bovine/Ohio/B24OSU-432/2024, H5N1, clade 2.3.4.4b), using four different assays including glycan array technology, bio-layer interferometry (BLI), solid-phase capture and haemagglutination of glycan-remodelled erythrocytes and found that it preferentially binds to ‘avian-type’ receptors (α2,3-sialosides). Although we find no evidence for human receptor specificity of influenza A/bovine virus, ongoing effort must be placed on continued monitoring for this trait.

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Fig. 1: Receptor-binding specificity of bovine H5N1.
Fig. 2: Binding of H5N1 and H1N1 viruses to glycan-remodelled erythrocytes.

Data availability

All data are available in the main text or the supplementary materials.

Code availability

The script for Microsoft Excel Macro for batch processing glycan microarray data is available at https://github.com/enthalpyliu/carbohydrate-microarray-processing.

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Acknowledgements

This research was funded in whole or in part with federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under Award Number R01 AI165692 (G.-J.B.) and contract numbers 75N93021C00016 (R.J.W.) and 75N93021C00018 (S.M.T.; NIAID Centers for Excellence for Influenza Research and Response, CEIRR). Funding was obtained from ICRAD and ERA-NET co-funded under the European Union’s Horizon 2020 research and innovation programme under grant agreement 862605 (R.P.d.V.)

Author information

Author notes

  1. These authors contributed equally: Pradeep Chopra, Sean D. Ray

Authors and Affiliations

  1. Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA

    Pradeep Chopra & Geert-Jan Boons

  2. Center for Influenza Disease and Emergence Response (CIDER), University of Georgia, Athens, GA, USA

    Pradeep Chopra, Sean D. Ray, Caroline K. Page, Justin D. Shepard, S. Mark Tompkins & Geert-Jan Boons

  3. Department of Infectious Diseases, University of Georgia, Athens, GA, USA

    Sean D. Ray, Caroline K. Page, Justin D. Shepard & S. Mark Tompkins

  4. Center for Vaccines and Immunology, University of Georgia, Athens, GA, USA

    Sean D. Ray, Caroline K. Page, Justin D. Shepard & S. Mark Tompkins

  5. Department of Infectious Diseases, St Jude Children’s Research Hospital, Memphis, TN, USA

    Ahmed Kandeil, Trushar Jeevan & Richard J. Webby

  6. St Jude Center of Excellence for Influenza Research and Response, Memphis, TN, USA

    Ahmed Kandeil, Trushar Jeevan, Andrew S. Bowman & Richard J. Webby

  7. Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University, Columbus, OH, USA

    Andrew S. Bowman

  8. Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO, USA

    Ali H. Ellebedy

  9. Center for Vaccines and Immunity to Microbial Pathogens, Washington University School of Medicine, St Louis, MO, USA

    Ali H. Ellebedy

  10. The Andrew M. and Jane M. Bursky Center for Human Immunology and Immunotherapy Programs, Washington University School of Medicine, St Louis, MO, USA

    Ali H. Ellebedy

  11. Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands

    Robert P. de Vries & Geert-Jan Boons

  12. Department of Chemistry, University of Georgia, Athens, GA, USA

    Geert-Jan Boons

Authors
  1. Pradeep Chopra
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Contributions

P.C., R.J.W., R.P.d.V., S.M.T. and G.-J.B. conceived the study. P.C., S.D.R., C.K.P., J.D.S., R.P.d.V., A.K. and T.J. performed the experiments. P.C., S.D.R., C.K.P., J.D.S., R.J.W., R.P.d.V., S.M.T. and G.-J.B. analysed the data and interpreted the results. A.S.B. isolated A/bovine/Ohio/B24OSU-432/2024 and A.H.E. developed reagents. P.C., S.D.R., S.M.T. and G.-J.B. wrote the manuscript. R.J.W., S.M.T. and G.-J.B. supervised research. All authors have given approval to the final version of the manuscript.

Corresponding authors

Correspondence to S. Mark Tompkins or Geert-Jan Boons.

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The authors declare no competing interests.

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Extended data figures and tables

Extended Data Fig. 1 Solid phase assays for receptor binding specificities.

The influenza A viruses (IAV) A/bovine/OH/B240SU-432/2024 (H5N1), A/Vietnam/1203/2004 (H5N1) and A/CA/04/2009 (pdmH1N1) (64 HA units) were captured on a fetuin-coated plate and incubated with 3’-SLN-PAA-biotin (10 µg/mL), 6’-SLN-PAA-biotin (10 µg/mL) or PBS in the presence or absence of neuraminidase inhibitor (Oseltamivir carboxylate, 2 μM). Virus-bound biotinylated polymer was detected using horseradish peroxidase (HRP)-conjugated streptavidin. The absorbance (A450) values for each treatment (in replicate of three, n = 3) of all viruses were plotted using Prism 10 software (GraphPad Software, Inc.). The assay was performed in triplicate at least two times for each virus, bars (scatter dot plot) represent the mean ± SEM (n = 3) for each treatment.

Source Data

Extended Data Table 1 HA receptor binding site (RBS) amino acid alignment of the H5 hemagglutinins
Full size table
Extended Data Table 2 HA titers of influenza A viruses with remodeled erythrocytes
Full size table

Supplementary information

Supplementary Information

Supplementary Information, including two supplementary figures, 1 supplementary table, and additional references.

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Supplementary Data

Source data for Supplementary Figs. 1 and 2.

Source data

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Chopra, P., Ray, S.D., Page, C.K. et al. Receptor-binding specificity of a bovine influenza A virus. Nature 640, E21–E27 (2025). https://doi.org/10.1038/s41586-025-08822-5

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