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Characterization of Influenza Virus Binding to Receptors on Isolated Cell Membranes

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Glycovirology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2556))

Abstract

An interplay between receptor-binding properties of influenza viruses (IVs) and spectrum of sialic acid-containing receptors on target cells in birds and mammals determine viral host range, tissue tropism, and pathogenicity. Here, we describe method that allows to characterize binding of IVs to biologically relevant cellular receptors using a conventional solid-phase enzyme-linked assay. In this method, we isolate plasma membranes from respiratory and intestinal epithelial cells of animal origin (Subheading 3.2). We adsorb the membranes in the wells of 96-well ELISA plates, incubate the membrane-coated wells with serially diluted IVs, and determine amounts of IVs attached to the membranes using viral ability to bind peroxidase-labeled sialoglycoprotein fetuin. Based on the concentration dependence of IV binding to the membrane, we estimate binding avidity and number of binding sites. We describe two variants of the assay in Subheadings 3.6 and 3.7 and provide examples.

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References

  1. Overeem NJ, van der Vries E, Huskens J (2021) A dynamic, supramolecular view on the multivalent interaction between influenza virus and host cell. Small 17:e2007214. https://doi.org/10.1002/smll.202007214

    Article  CAS  PubMed  Google Scholar 

  2. Ji Y, White YJ, Hadden JA et al (2017) New insights into influenza A specificity: an evolution of paradigms. Curr Opin Struct Biol 44:219–231. https://doi.org/10.1016/j.sbi.2017.06.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Klenk HD, Garten W, Matrosovich M (2013) Pathogenesis. In: Webster RG (ed) Textbook of influenza. John Wiley & Sons, Ltd, Chichester, pp 157–172

    Chapter  Google Scholar 

  4. Kuchipudi SV, Nelli RK, Gontu A et al (2021) Sialic acid receptors: the key to solving the enigma of zoonotic virus spillover. Viruses 13(2):262. https://doi.org/10.3390/v13020262

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Matrosovich MN, Klenk H-D, Kawaoka Y (2006) Receptor specificity, host range and pathogenicity of influenza viruses. In: Kawaoka Y (ed) Influenza virology: current topics. Caister Academic Press, Wymondham, pp 95–137

    Google Scholar 

  6. Xiong X, McCauley JW, Steinhauer DA (2014) Receptor binding properties of the influenza virus hemagglutinin as a determinant of host range. In: Compans R, Oldstone M (eds) Influenza pathogenesis and control – volume I, Current topics in microbiology and immunology, vol 385. Springer, Cham, pp 63–91. https://doi.org/10.1007/82_2014_423

    Chapter  Google Scholar 

  7. Matrosovich MN, Gambaryan AS (2012) Solid-phase assays of receptor-binding specificity. In: Kawaoka Y, Neumann G (eds) Influenza virus. Methods in molecular biology (methods and protocols), vol 865. Humana Press, pp 71–94. https://doi.org/10.1007/978-1-61779-621-0_5

    Chapter  Google Scholar 

  8. Sriwilaijaroen N, Suzuki Y (2020) Host receptors of influenza viruses and coronaviruses--molecular mechanisms of recognition. Vaccine 8(4):587. https://doi.org/10.3390/vaccines8040587

    Article  CAS  Google Scholar 

  9. van Riel D, Munster VJ, de Wit E et al (2007) Human and avian influenza viruses target different cells in the lower respiratory tract of humans and other mammals. Am J Pathol 171(4):1215–1223. https://doi.org/10.2353/ajpath.2007.070248

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Verhagen JH, Eriksson P, Leijten L et al (2021) Host range of influenza A virus H1 to H16 in Eurasian ducks based on tissue and receptor binding studies. J Virol 95(6):e01873–e01820. https://doi.org/10.1128/JVI.01873-20

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Eriksson P, Lindskog C, Lorente-Leal V et al (2019) Attachment patterns of human and avian influenza viruses to trachea and colon of 26 bird species – support for the community concept. Front Microbiol 10:815. https://doi.org/10.3389/fmicb.2019.00815

    Article  PubMed  PubMed Central  Google Scholar 

  12. Detmer SE, Gramer MR, Goyal SM et al (2013) In vitro characterization of influenza A virus attachment in the upper and lower respiratory tracts of pigs. Vet Pathol 50(4):648–658. https://doi.org/10.1177/0300985812467469

    Article  CAS  PubMed  Google Scholar 

  13. Wang B, Liu B, Chen L et al (2012) Qualitative and quantitative analyses of influenza virus receptors in trachea and lung tissues of humans, mice, chickens and ducks. Sci China Life Sci 55(7):612–617. https://doi.org/10.1007/s11427-012-4341-8

    Article  CAS  PubMed  Google Scholar 

  14. Wickramasinghe IN, de Vries RP, Eggert AM et al (2015) Host tissue and glycan binding specificities of avian viral attachment proteins using novel avian tissue microarrays. PLoS One 10(6):e0128893. https://doi.org/10.1371/journal.pone.0128893

    Article  CAS  PubMed  Google Scholar 

  15. Matrosovich M, Matrosovich T, Carr J et al (2003) Overexpression of the alpha-2,6-sialyltransferase in MDCK cells increases influenza virus sensitivity to neuraminidase inhibitors. J Virol 77(15):8418–8425. https://doi.org/10.1128/JVI.77.15.8418-8425.2003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Govorkova EA, Matrosovich MN, Tuzikov AB et al (1999) Selection of receptor-binding variants of human influenza A and B viruses in baby hamster kidney cells. Virology 262(1):31–38. https://doi.org/10.1006/viro.1999.9892

    Article  CAS  PubMed  Google Scholar 

  17. Gubareva LV, Matrosovich MN, Brenner MK et al (1998) Evidence for zanamivir resistance in an immunocompromised child infected with influenza B virus. J Infect Dis 178(5):1257–1262. https://doi.org/10.1086/314440

    Article  CAS  PubMed  Google Scholar 

  18. Gambaryan AS, Matrosovich MN, Bender CA et al (1998) Differences in the biological phenotype of low-yielding (L) and high-yielding (H) variants of swine influenza virus A/NJ/11/76 are associated with their different receptor-binding activity. Virology 247(2):223–231. https://doi.org/10.1006/viro.1998.9274

    Article  CAS  PubMed  Google Scholar 

  19. Gambaryan AS, Marinina VP, Tuzikov AB et al (1998) Effects of host-dependent glycosylation of hemagglutinin on receptor-binding properties on H1N1 human influenza A virus grown in MDCK cells and in embryonated eggs. Virology 247(2):170–177. https://doi.org/10.1006/viro.1998.9224

    Article  CAS  PubMed  Google Scholar 

  20. Gambaryan AS, Yamnikova SS, Lvov DK et al (2002) Differences in receptor specificity between the influenza viruses of duck, chicken, and human. Mol Biol 36(3):429–435. https://doi.org/10.1023/A:1016023700050

    Article  CAS  Google Scholar 

  21. Gambaryan A, Webster R, Matrosovich M (2002) Differences between influenza virus receptors on target cells of duck and chicken. Arch Virol 147(6):1197–1208. https://doi.org/10.1007/s00705-002-0796-4

    Article  CAS  PubMed  Google Scholar 

  22. Gambaryan AS, Tuzikov AB, Pazynina GV et al (2004) H5N1 chicken influenza viruses display a high binding affinity for Neu5Ac alpha 2-3Gal beta 1-4(6-HSO3)GlcNAc-containing receptors. Virology 326(2):310–316. https://doi.org/10.1016/j.virol.2004.06.002

    Article  CAS  PubMed  Google Scholar 

  23. Gambarian AS, Marinina VP, Solodar TA et al (2006) Different receptor specificity of influence viruses from ducks and chickens and its reflection in the composition of sialosides on host cells and mucins. Vopr Virusol 51(4):24–32

    CAS  PubMed  Google Scholar 

  24. Szretter KJ, Balish AL, Katz JM (2006) Influenza: propagation, quantification, and storage. Curr Protoc Microbiol 3(1):15G. https://doi.org/10.1002/0471729256.mc15g01s3

    Article  Google Scholar 

  25. World Health Organization (2002) WHO manual on animal influenza diagnosis and surveillance. No. WHO/CDS/CSR/NCS/2002.5

    Google Scholar 

  26. Eisfeld AJ, Neumann G, Kawaoka Y (2014) Influenza A virus isolation, culture and identification. Nat Protoc 9(11):2663–2681. https://doi.org/10.1038/nprot.2014.180

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Yoon SW, Webby RJ, Webster RG (2014) Evolution and ecology of influenza A viruses. In: Compans R, Oldstone M (eds) Influenza pathogenesis and control – volume I, Current topics in microbiology and immunology, vol 385. Springer, Cham, pp 359–375. https://doi.org/10.1007/82_2014_396

    Chapter  Google Scholar 

  28. Gambaryan AS, Matrosovich MN (1992) A solid-phase enzyme-linked assay for influenza virus receptor-binding activity. J Virol Methods 39(1–2):111–123. https://doi.org/10.1016/0166-0934(92)90130-6

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We acknowledge recent financial support of our work by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Project number 197785619—SFB 1021 and by the Russian Foundation for Basic Research—research grants 14-04-00547 and 17-04-00148.

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Authors and Affiliations

  1. Institute of Virology, Philipps University, Marburg, Germany

    Mikhail N. Matrosovich

  2. Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products, Russian Academy of Sciences, Moscow, Russia

    Alexandra S. Gambaryan

Authors
  1. Mikhail N. Matrosovich
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  2. Alexandra S. Gambaryan
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Editors and Affiliations

  1. Department of Medical Biochemistry, University of Shizuoka School of Pharmaceutical Sciences, Shizuoka, Japan

    Yasuo Suzuki

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Matrosovich, M.N., Gambaryan, A.S. (2022). Characterization of Influenza Virus Binding to Receptors on Isolated Cell Membranes. In: Suzuki, Y. (eds) Glycovirology. Methods in Molecular Biology, vol 2556. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2635-1_12

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  • DOI: https://doi.org/10.1007/978-1-0716-2635-1_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2634-4

  • Online ISBN: 978-1-0716-2635-1

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