Humanized cell line
AssigneesWisconsin Alumni Research Foundation (WARF), The University of Tokyo
InventorsYoshihiro Kawaoka, Kosuke Takada, Masaki Imai
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Abstract
A mammalian or avian cell line that expresses high levels of human influenza virus receptors is provided. In one embodiment, the cell line supports human influenza virus, e.g., human A/H3 influenza virus, isolation and growth much more effectively than corresponding conventional (unmodified) cells or in corresponding human virus receptor-overexpressing cells, and the propagated viruses may maintain higher genetic stability than in the corresponding cells.
Claims19
An isolated recombinant mammalian cell, comprising a reduced amount of cell surface β-galactoside α2,3 sialyl residues relative to a corresponding non-recombinant mammalian cell, wherein β-galactoside α2,3 sialyltransferase (ST3Gal) genes are mutated so as to reduce the amount of the cell surface β-galactoside α2,3 sialyl residues, wherein the ST3Gal genes include ST3Gal-I, ST3Gal-II, ST3Gal-III, ST3Gal-IV, ST3Gal-V, ST3Gal-VI, and ST3Gal-II-like genes, and wherein the recombinant mammalian cell comprises an expression cassette encoding human β-galactoside α2,6 sialyltransferase (ST6Gal), wherein the reduced amount of cell surface 3-galactoside a2,3 sialyl residues is the result of mutation of the ST3Gal genes in the recombinant cell.
A method of modifying the amount of cell surface β-galactoside α2,3 sialyl residues and human β-galactoside α2,6 sialyl residues on a mammalian cell, comprising:
- mutating β-galactoside α2,3 sialyltransferase (ST3Gal) genes, wherein the ST3Gal genes include ST3Gal-I, ST3Gal-II, ST3Gal-III, ST3Gal-IV, ST3Gal-V, ST3Gal-VI, and ST3Gal-II-like genes, and
- overexpressing a human β-galactoside α2,6 sialyltransferase (ST6Gal) gene, in a parental mammalian cell so as to result in a modified mammalian cell having a reduced amount of cell surface β-galactoside α2,3 sialyl residues and an increased amount of human p β-galactoside α2,6 sialyl residues on the surface of the modified cell relative to the corresponding parental mammalian cell.
Dependencies are inferred from the claim text.