Patent ID: 8057806
Filing Date: 2011-11-15
Classification: A61K,A61P,C07K,C12N

Abstract:
1. A method of preparing an attenuated recombinant influenza virus comprising a mutant M2 protein gene, comprising: (i) transfecting an isolated a host cell with a plurality of influenza virus vRNA and influenza virus protein encoding vectors so as to yield recombinant influenza virus, wherein the plurality of vectors comprises: a) influenza virus vRNA vectors comprising a vector comprising a promoter operably linked to an influenza virus PA cDNA linked to a transcription termination sequence, a vector comprising a promoter operably linked to an influenza virus PB1 cDNA linked to a transcription termination sequence, a vector comprising a promoter operably linked to an influenza virus PB2 cDNA linked to a transcription termination sequence, a vector comprising a promoter operably linked to an influenza virus HA cDNA linked to a transcription termination sequence, a vector comprising promoter operably linked to an influenza virus NP cDNA linked to a transcription termination sequence, a vector comprising a promoter operably linked to an influenza virus NA cDNA linked to a transcription termination sequence, a vector comprising a promoter operably linked to an influenza virus M cDNA linked to a transcription termination sequence, and a vector comprising a promoter operably linked to an influenza virus NS cDNA linked to a transcription termination sequence, wherein the M cDNA comprises a mutation in the transmembrane domain of M2 protein DNA, wherein the mutation is a deletion of residues that include residues between residues 29 to 31, and wherein the mutation is associated with attenuation of the virus in vivo; and b) influenza virus protein encoding vectors comprising a vector comprising a promoter operably linked to a DNA segment encoding influenza virus PA, a vector comprising a promoter operably linked to a DNA segment encoding influenza virus PB1, a vector comprising a promoter operably linked to a DNA segment encoding influenza virus PB2, and a vector comprising a promoter operably linked to a DNA segment encoding influenza virus NP, and optionally a vector comprising a promoter operably linked to a DNA segment encoding influenza virus HA, a vector comprising a promoter operably linked to a DNA segment encoding influenza virus NA, a vector comprising a promoter operably linked to a DNA segment encoding influenza virus M1, a vector comprising a promoter operably linked to a DNA segment encoding an influenza A virus ion channel protein, and a vector comprising a promoter operably linked to a DNA segment encoding influenza virus NS2; and (ii) isolating the virus.