Recombinant tryptophan mutants of influenza
AssigneeAviron
InventorNeil T. Parkin
Life of this patent
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Abstract
Recombinant PB2 tryptophan variant influenza viruses, RNA, cDNA and vectors are provided. Also provided are immunogenic compositions containing the variant viruses, methods of producing such viruses and methods for the prophylactic treatment of influenza in humans.
Claims22
A method of making a recombinant influenza virus in which the amino acid sequence comprising the PB2 protein has been modified by the substitution of at least one non-native amino acid for at least one native amino acids that affects the cap-binding activity of PB2 in the region spanning amino acid residues 537 through 575 comprising:
- (i) introducing a negative-strand RNA sequence encoding the modified PB2 protein into host cells infected with helper virus capable of producing influenza virus RNA segments;
- (ii) recovering from the host cells viruses that have incorporated into their genomes the introduced recombinant, negative-strand RNA sequence.
A method of making a recombinant influenza virus in which the amino acid sequence comprising the PB2 protein has been modified by the substitution of at least one non-native amino acid for at least one native amino acids that affects the cap-binding activity of PB2 in the region spanning amino acid residues 537 through 575 comprising:
- (i) infecting host cells with a helper virus capable of producing influenza virus RNA segments;
- (ii) generating cDNA encoding the modified PB2 protein;
- (iii) preparing a vRNA-sense RNP complex by transcription of the cDNA with a polymerase enzyme;
- (iv) electroporating the host cells with the RNP mixture;
- and
- (v) isolating the recombinant influenza virus from the host cells.
A method for the treatment of humans for influenza infection, comprising administering to a human patient in need of treatment of immunologically inducing effective amount of a composition composed of recombinant influenza virus in admixture with pharmaceutically acceptable carrier, the recombinant influenza virus containing RNA encoding an amino acid sequence comprising a PB2 protein modified by substitution of at least one-non-native amino acid for a native tryptophan residue that affects cap binding activity of PB2 in the region spanning amino acid residues 537 through 575.
Dependencies are inferred from the claim text.