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 () and James C. Paulson ()
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Chika Kikuchi: The Scripps Research Institute
Aristotelis Antonopoulos: Imperial College London
Shengyang Wang: The Scripps Research Institute
Tadashi Maemura: University of Wisconsin-Madison
Rositsa Karamanska: Imperial College London
Chiara Lee: Imperial College London
Andrew J. Thompson: The Scripps Research Institute
Anne Dell: Imperial College London
Yoshihiro Kawaoka: University of Wisconsin-Madison
Stuart M. Haslam: Imperial College London
James C. Paulson: The Scripps Research Institute

Nature Communications, 2023, vol. 14, issue 1, 1-13

Abstract: 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.

Date: 2023
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DOI: 10.1038/s41467-023-41908-0

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