Glyco-engineered MDCK cells display preferred receptors of H3N2 influenza absent in eggs used for vaccines

Chika Kikuchi; Aristotelis Antonopoulos; Shengyang Wang; Tadashi Maemura; Rositsa Karamanska; Chiara Lee; Andrew J Thompson; Anne Dell; Yoshihiro Kawaoka; Stuart M Haslam; James C Paulson

doi:10.1038/s41467-023-41908-0

Glyco-engineered MDCK cells display preferred receptors of H3N2 influenza absent in eggs used for vaccines

Nat Commun. 2023 Oct 4;14(1):6178. doi: 10.1038/s41467-023-41908-0.

Authors

Chika Kikuchi^#^{1

2}, Aristotelis Antonopoulos^#³, Shengyang Wang^{1

2}, Tadashi Maemura⁴, Rositsa Karamanska³, Chiara Lee³, Andrew J Thompson^{1

2}, Anne Dell³, Yoshihiro Kawaoka^{4

5

6

7}, Stuart M Haslam⁸, James C Paulson^{9

10}

Affiliations

¹ Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, USA.
² Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA.
³ Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK.
⁴ Influenza Research Institute, Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI, USA.
⁵ Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, The University of Tokyo, Tokyo, Japan.
⁶ The Research Center for Global Viral Diseases, National Center for Global Health and Medicine Research Institute, Tokyo, Japan.
⁷ Pandemic Preparedness, Infection and Advanced Research Center, The University of Tokyo, Tokyo, Japan.
⁸ Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK. s.haslam@imperial.ac.uk.
⁹ Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA, USA. jpaulson@scripps.edu.
¹⁰ Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA, USA. jpaulson@scripps.edu.

^# Contributed equally.

Abstract

Evolution of human H3N2 influenza viruses driven by immune selection has narrowed the receptor specificity of the hemagglutinin (HA) to a restricted subset of human-type (Neu5Acα2-6 Gal) glycan receptors that have extended poly-LacNAc (Galβ1-4GlcNAc) repeats. This altered specificity has presented challenges for hemagglutination assays, growth in laboratory hosts, and vaccine production in eggs. To assess the impact of extended glycan receptors on virus binding, infection, and growth, we have engineered N-glycan extended (NExt) cell lines by overexpressing β3-Ν-acetylglucosaminyltransferase 2 in MDCK, SIAT, and hCK cell lines. Of these, SIAT-NExt cells exhibit markedly increased binding of H3 HAs and susceptibility to infection by recent H3N2 virus strains, but without impacting final virus titers. Glycome analysis of these cell lines and allantoic and amniotic egg membranes provide insights into the importance of extended glycan receptors for growth of recent H3N2 viruses and relevance to their production for cell- and egg-based vaccines.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Dogs
Hemagglutinin Glycoproteins, Influenza Virus
Humans
Influenza A Virus, H3N2 Subtype
Influenza Vaccines*
Influenza, Human* / prevention & control
Madin Darby Canine Kidney Cells
Polysaccharides / metabolism

Substances

Influenza Vaccines
Polysaccharides
Hemagglutinin Glycoproteins, Influenza Virus

Abstract

Publication types

MeSH terms

Substances

Grants and funding