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Composition génétique de semences vaccinales H3N2 et construction d'un virus vecteur : une histoire d'encapsidation de segments chez les virus influenza de type A

Abstract : The packaging of the eight segments corresponding to the influenza A viruses genomeis a key process of the viral replication as well as a stake of actual scientific researchesbecause it leads to reassortant viruses, e.g. pandemic viruses. We studied the two main facetsof influenza segment packaging: reassortment, during vaccine seeds production and foreignsegment incorporation for influenza vector construction. Vaccine seeds are produced bycoinfection of hens’ eggs with two viruses, a donor one (reference circulating strain) and anacceptor one (A/Puerto Rico/8/34 (H1N1) (PR8)). Analysis of internal genetic composition ofthirteen H3N2 vaccine seeds reveals that PB1 segment of H3N2 donor strain is incorporatedin more than fifty per cent of the cases. Moreover, coinfection events lead to an extremelywide range of reassortants from 6:2 to 2:6 (PR8:H3N2). Segment incorporation competitionassays performed using plasmid-based reverse genetics show that selective packaging of PB1segment is based on genetic environment, i.e. viral origin of HA and NA segments. Thesecond part of my PhD work has been devoted to replicative influenza vector based on H3virus isolated from patients without NA segment at the native stage. None of the gfptransgenic constructions containing reporter gene have been incorporated in viral particles,contrary to literature studies performed using H1N1 laboratory-adapted strains. Even ifmolecular mechanisms controlling influenza A viruses segments incorporation remain stillcomplex, genetic background seems to be an essential element which must be considered withinterest.
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Corinne Bergeron. Composition génétique de semences vaccinales H3N2 et construction d'un virus vecteur : une histoire d'encapsidation de segments chez les virus influenza de type A. Sciences agricoles. Université Claude Bernard - Lyon I, 2009. Français. ⟨NNT : 2009LYO10332⟩. ⟨tel-00625467⟩

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