Recombinant viruses expressing alpha-1, 3-galactosyltransferase and uses thereof
Disclosed are viruses, and vaccines comprised of and made from such viruses, that include a heterologous nucleic acid segment encoding α-1,3-galactosyltransferase (α-1,3-GT) such that the nucleic acid segment expresses α-1,3-GT when the virus infects a host cell. Such viruses produce proteins having α-1,3-galactose. The presence of α-1,3-galactose on proteins of infected cells can powerfully stimulate the immune response of the host against the viral proteins of the virus, thus enhancing the effect of the virus as a vaccine. Also disclosed are vaccines that include and/or are produced by such viruses. Also disclosed are methods of making and using such viruses and vaccines, such as administering to a subject in need thereof a vaccine as disclosed and such as making a vaccine that includes one or more viral proteins expressed by a virus as disclosed.
1 . An attenuated live influenza virus comprising a heterologous nucleic acid encoding α-1,3-galactosyltransferase (α-1,3-GT),
wherein, upon infecting a host cell the attenuated live influenza virus expresses α-1,3-GT,
wherein the expressed α-1,3-GT synthesizes α-1,3-galactose in the host cell, and
wherein the α-1,3-galactose is exposed on the surface of the host cell.
2 . The attenuated live influenza virus of claim 1 , wherein the heterologous nucleic acid encoding α-1,3-GT is fused to a nucleic acid encoding at least one viral protein expressed by the attenuated live influenza virus.
3 . The attenuated live influenza virus of claim 2 , wherein the heterologous nucleic acid encoding α-1,3-GT is fused to the nucleic acid encoding the viral protein expressed by the attenuated live influenza virus via one or more linkers.
4 . The attenuated live influenza virus of claim 3 , wherein the one or more linkers are peptide linkers.
5 . The attenuated live influenza virus of claim 2 , further comprising a nucleic acid encoding a 2A self-cleaving peptide between the heterologous nucleic acid encoding α-1,3-GT and the nucleic acid encoding the viral protein.
6 . The attenuated live influenza virus of claim 5 , wherein the nucleic acid encoding the 2A self-cleaving peptide comprises a protease cleavage site.
7 . The attenuated live influenza virus of claim 5 , wherein the 2A self-cleaving peptide is derived from porcine teschovirus-1.
8 . The attenuated live influenza virus of claim 5 , wherein the 2A self-cleaving peptide is flanked by short peptide linkers.
9 . The attenuated live influenza virus of claim 8 , wherein the short peptides linkers are GSG.
10 . The attenuated live influenza virus of claim 2 , wherein the viral protein is neuraminidase (NA).
11 . The virus of claim 1 , wherein the nucleic acid encoding α-1,3-GT is incorporated into the genome of the attenuated live influenza virus.
12 . The attenuated live influenza virus of claim 1 , wherein the heterologous nucleic acid encoding α-1,3-GT is a mammalian nucleic acid encoding α-1,3-GT.
13 . The attenuated live influenza virus of claim 1 , wherein the heterologous nucleic acid encoding α-1,3-GT is a mouse nucleic acid encoding α-1,3-GT.
14 . The attenuated live influenza virus of claim 1 , wherein the virus is attenuated by serial passage in cultured cells, serial passage in a heterologous host animal, gene deletion in the virus, site-directed mutagenesis of the virus, altering codon usage of the virus, selection of cold-adapted mutants, using a related virus from a heterologous host species, using a naturally occurring attenuated virus strain, or combinations thereof.
15 . The virus of claim 14 , wherein the virus is attenuated by serial passage in cultured cells, wherein the cultured cells are Madin-Darby Canine Kidney (MDCK) cells.
16 . A vaccine comprising the virus of claim 1 .
17 . A method of vaccination, the method comprising administering to a subject in need thereof a vaccine of claim 16 .
18 . The method of claim 17 , wherein the vaccine is administered intranasally, pulmonarily, orally, subcutaneously, intramuscularly, intradermally, or intraperitoneally.
19 . A vaccine comprising one or more viral proteins expressed by the virus of claim 1 , wherein the one or more viral proteins comprise α-1,3-galactose.
20 . A method of making the vaccine of claim 19 , the method comprising infecting a cell with the virus, whereby the one or more viral proteins comprising α-1,3-galactose are produced.
21 . The method of claim 20 further comprising purifying the one or more viral proteins.