| Title: |
Contemporary human H3N2 influenza a viruses require a low threshold of suitable glycan receptors for efficient infection |
| Authors: |
Spruit, Cindy M; Sweet, Igor R; Maliepaard, Joshua C L; Bestebroer, Theo; Lexmond, Pascal; Qiu, Boning; Damen, Mirjam J A; Fouchier, Ron A M; Reiding, Karli R; Snijder, Joost; Herfst, Sander; Boons, Geert-Jan; de Vries, Robert P; Afd Chemical Biology and Drug Discovery; Afd Biomol.Mass Spect. and Proteomics; Afd Pharmaceutics; Sub Biomol.Mass Spectrometry & Proteom.; Sub Chemical Biology and Drug Discovery; Chemical Biology and Drug Discovery; Biomolecular Mass Spectrometry and Proteomics; Pharmaceutics |
| Publication Year: |
2023 |
| Subject Terms: |
sialic acid; poly-LacNAc; influenza; H3N2; genetic glycoengineering |
| Description: |
Recent human H3N2 influenza A viruses have evolved to employ elongated glycans terminating in α2,6-linked sialic acid as their receptors. These glycans are displayed in low abundancies by (humanized) Madin-Darby Canine Kidney cells, which are commonly employed to propagate influenza A virus, resulting in low or no viral propagation. Here, we examined whether the overexpression of the glycosyltransferases β-1,3-N-acetylglucosaminyltransferase and β-1,4-galactosyltransferase 1, which are responsible for the elongation of poly-N-acetyllactosamines (LacNAcs), would result in improved A/H3N2 propagation. Stable overexpression of β-1,3-N-acetylglucosaminyltransferase and β-1,4-galactosyltransferase 1 in Madin-Darby Canine Kidney and "humanized" Madin-Darby Canine Kidney cells was achieved by lentiviral integration and subsequent antibiotic selection and confirmed by qPCR and protein mass spectrometry experiments. Flow cytometry and glycan mass spectrometry experiments using the β-1,3-N-acetylglucosaminyltransferase and/or β-1,4-galactosyltransferase 1 knock-in cells demonstrated increased binding of viral hemagglutinins and the presence of a larger number of LacNAc repeating units, especially on "humanized" Madin-Darby Canine Kidney-β-1,3-N-acetylglucosaminyltransferase cells. An increase in the number of glycan receptors did, however, not result in a greater infection efficiency of recent human H3N2 viruses. Based on these results, we propose that H3N2 influenza A viruses require a low number of suitable glycan receptors to infect cells and that an increase in the glycan receptor display above this threshold does not result in improved infection efficiency. |
| Document Type: |
article in journal/newspaper |
| File Description: |
text/plain |
| Language: |
English |
| ISSN: |
0959-6658 |
| Relation: |
https://dspace.library.uu.nl/handle/1874/433297 |
| Availability: |
https://dspace.library.uu.nl/handle/1874/433297 |
| Rights: |
info:eu-repo/semantics/OpenAccess |
| Accession Number: |
edsbas.207E43D8 |
| Database: |
BASE |