Temperature sensitive mutants of influenza A virus generated by reverse genetics and clustered charged to alanine mutagenesis

Temperature sensitive ( ts) mutants of influenza A virus have the potential to serve as live attenuated ( att) virus vaccines. Previously, ts mutants were isolated by chemical mutagenesis or arose spontaneously, and most likely contained point mutations in one or more genes. While sufficiently atten...

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Bibliographic Details
Published in:Virus research 1996-12, Vol.46 (1), p.31-44
Main Authors: Parkin, Neil T., Chiu, Peggy, Coelingh, Kathleen L.
Format: Article
Language:English
Subjects:
Alanine - genetics
Alanine mutagenesis
Animals
Dogs
Ferrets
Helper Viruses - genetics
Helper Viruses - growth & development
Influenza
influenza A virus
Influenza A virus - genetics
Influenza A virus - growth & development
Kidney
Mice
Mice, Inbred BALB C
Molecular Sequence Data
Mutagenesis
PB2
Phenotype
Temperature
Temperature sensitive
Transfection
Virus Replication - genetics
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Summary:Temperature sensitive ( ts) mutants of influenza A virus have the potential to serve as live attenuated ( att) virus vaccines. Previously, ts mutants were isolated by chemical mutagenesis or arose spontaneously, and most likely contained point mutations in one or more genes. While sufficiently attenuated, even the most genetically stable of these viruses was found to revert to a more virulent form in a seronegative vaccinee. Recently developed technology, however, allows the introduction of engineered mutations into the genome of influenza A and B viruses, permitting the rational design of attenuated mutants with the potential for increased genetic stability. To accomplish this goal, we have introduced ts mutations into the PB2 gene of A/Los Angeles/2/87 (H3N2) and rescued the mutated genes into infectious viruses. We have used clustered charged to alanine mutagenesis (substitution of alanine for charged amino acid residues which are present in clusters) of the PB2 gene to generate novel ts mutants. Viruses containing such ts PB2 genes were attenuated in mice and ferrets. This approach has thus yielded several vaccine candidates with ts and attenuated characteristics in animal models. Combination of these mutations with each other or with other ts mutations may lead to a high level of genetic stability.
ISSN:0168-1702
1872-7492
DOI:10.1016/S0168-1702(96)01373-1