HEPATITIS C VIRUS EXPRESSING REPORTER TAGGED NS5A PROTEIN

The present inventors developed hepatitis C reporter viruses containing Core through NS2 of prototype isolates of all major HCV genotypes and the remaining genes of isolate JFH1, by insertion of reporter genes in domain III of HCV NS5A. The inventors have identified a deletion upstream of the inserted reporter gene sequence, which conferred favourable growth kinetics in Huh7.5 cells to these viruses. These reporter viruses can be used for high throughput analysis of drug and vaccine candidates as well as patient samples. Drugs could be evaluated for their potential to prevent infection or cure infected cells. The neutralizing capacity of antibodies induced by vaccine candidates could be evaluated in order to define successful vaccination strategies. Broadly neutralizing antibodies could be identified testing engineered antibodies and antibodies derived from serum of HCV infected individuals; thus this technique could contribute to the development of immunotherapy. The developed systems could aid individualized treatment of HCV infected: Patient isolates could be tested for resistance to drugs by introduction of genome regions involved in drug resistance in the developed constructs and subsequent treatment with the drug of interest. The present inventorsalso developed JFH1-based intergenotypic recombinants with genotype specific homotypic 5UTR, or heterotypic 5'UTR (either of genotype 1a (strain H77) or of genotype 3a (strain S52)). The present inventors additionallydeveloped J6/JFH1 recombinants with the 5'UTR of genotypes 1-6. These recombinants with different 5UTRs are a useful to study the function of the 5'UTR in a genotype specific manner. Les présents inventeurs ont développé des virus rapporteurs de l'hépatite C contenant les protéines du noyau jusqu'à NS2 d'isolats prototypes de tous les principaux génotypes du VHC et les gènes restants d'isolat JFH1, par insertion de gènes rapporteurs dans le domaine III de NS5A du VHC. Les inventeurs ont identifié une délétion en amont de la séquence du gène rapporteur inséré, qui conférait une cinétique de croissance favorable dans des cellules Huh7.5 à ces virus. Ces virus rapporteurs peuvent être utilisés pour l'analyse à haut débit de médicaments et vaccins candidats ainsi que d'échantillons de patients. Des médicaments pourraient être évalués quant à leur potentiel à prévenir une infection ou à guérir des cellules infectées. L'aptitude de neutralisation d'anticorps induits par des vaccins candidats pourrait