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.
Claims17
An isolated recombinant canine cell, comprising a reduced amount of cell surface β-galactoside α2,3 sialyl residues relative to a corresponding non-recombinant canine cell, wherein seven different β-galactoside α2,3 sialyltransferase (ST3) genes are mutated so as to reduce the amount of the cell surface β-galactoside α2,3 sialyl residues, wherein the seven different genes include ST3Gal-I, ST3Gal-II, ST3Gal-III, ST3Gal-IV, ST3Gal-V, ST3Gal-VI, and ST3Gal-II-like genes, and wherein the recombinant canine cell comprises an expression cassette encoding human β-galactoside α2,6 sialyltransferase I (ST6Gal-I) or ST6Gal-II, wherein the reduced amount of cell surface 3-galactoside α2,3 sialyl residues is the result of recombinase knock out mutation of the plurality of ST3 genes in the recombinant cell, and wherein the recombinant canine cell provides for increased human influenza virus replication relative to the corresponding non-recombinant canine 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 canine cell, comprising:
- mutating seven different β-galactoside α2,3 sialyltransferase (ST3Gal) genes, wherein the seven different 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 canine cell so as to result in a modified canine cell having a reduced amount of cell surface β-galactoside α2,3 sialyl residues and an increased amount of human β-galactoside α2,6 sialyl residues on the surface of the modified cell relative to the corresponding parental canine cell.
An isolated recombinant MDCK cell comprising a reduced amount of cell surface β-galactoside α2,3 sialyl residues relative to a parental MDCK cell, wherein seven β-galactoside α2,3 sialyltransferase (ST3) genes are mutated so as to reduce the amount of the cell surface β-galactoside α2,3 sialyl residues, and wherein the recombinant MDCK cell comprises an expression cassette encoding human β-galactoside α2,6 sialyltransferase I (ST6Gal-I) or ST6Gal-II, wherein the reduced amount of cell surface β-galactoside α2,3 sialyl residues is the result of recombinase knock out mutation of ST3Gal-I, ST3Gal-II, ST3Gal-III, ST3Gal-IV, ST3Gal-V, ST3Gal-VI, and ST3Gal-II-like genes in the recombinant MDCK cell.
Dependencies are inferred from the claim text.