Attenuated vaccination and gene-transfer virus, a method to make the virus and a pharmaceutical composition comprising the virus

US6524588B1 · kind B1 · utility

Assignee

• Gerd Hobom · DE

Inventors

• Gerd Hobom · Neu-Ulm, DE
• Gabriele Neumann · Madison, US
• Annette Menke · Marburg, DE

Key dates

• Filing date: Mar 24, 1997
• Grant date: Feb 25, 2003
• Expiry date: Mar 24, 2017

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2760/16161
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

RNA polymerase I transcription in vivo in transiently DNA-transfected cells has been used for expression of influenza vRNA molecules coding for chloramphenicol acetyltransferase (CAT) in anti-sense orientation. Influenza virus superinfection served to provide viral RNA polymerase and other proteins for transcriptional conversion of minus-strand vRNA into plus-strand viral mRNA molecules expressing CAT activity. This system has been used for an analysis via nucleotide exchanges as well as deletions and insertions of both terminal segments of the vRNA sequence which cooperatively constitute the vRNA promoter structure. Several mutants with greatly enhanced expression rates over wild-type levels have been constructed, which also can be packaged and serially passaged into progeny virus. The data obtained for the mutations in various promoter elements support a model of consecutive, double strand vRNA promoter structures in binding of viral polymerase and initiation of RNA synthesis. Preparations of attenuated influenza virus for vaccination purposes include a single recombinant segment with promoter up mutation(s) for over-expression of an own or foreign gene product, which at the same…