US 7,402,429 B1
Host cells comprising cDNA corresponding to the antigenome of nonsegmented negative strand RNA viruses, further encoding antigenically active proteins
Martin A. Billeter, Zurich (Switzerland); Pius Spielhofer, Zurich (Switzerland); Karin Kälin, Gif-sur-Yvette (France); Frank Radecke, Zurich (Switzerland); and Henriette Schneider, Zurich (Switzerland)
Assigned to Schweiz Serum & Impfinstitut Bern, (Switzerland)
Appl. No. 9/11,425
PCT Filed Aug. 09, 1996, PCT No. PCT/EP96/03544
§ 371(c)(1), (2), (4) Date Sep. 15, 1998,
PCT Pub. No. WO97/06270, PCT Pub. Date Feb. 20, 1997.
Claims priority of application No. 95112559 (EP), filed on Aug. 09, 1995.
Int. Cl. A01N 63/00 (2006.01); A01N 65/00 (2006.01); A61K 48/00 (2006.01); C12N 5/00 (2006.01); C12N 5/02 (2006.01); C12N 15/00 (2006.01)
U.S. Cl. 435—325  [435/320.1; 424/93.1; 424/93.2] 20 Claims
 
1. An isolated eukaryotic host cell comprising (1) stably transfected expression plasmids that allow genomic expression of an exogenous RNA polymerase, a viral N protein, and a viral P protein in the host cell; (2) a nucleic acid sequence encoding a viral L protein; and (3) a cDNA molecule, said cDNA molecule comprising:
(a) a (+) strand sequence of a non-segmented negative-strand RNA virus of the family Paramyxoviridae from which RNA transcripts bearing the authentic 3′ termini can be transcribed; and
(b) an expression control sequence,
wherein said (+) strand sequence of a non-segmented negative-strand RNA virus is operatively linked to the expression control sequence, and wherein the host cell, without a replicating vaccinia helper virus, produces an infectious RNA virus containing the (−) strand sequence of the non-segmented negative-strand RNA virus of the family Paramyxoviridae, said (−) strand sequence of the non-segmented negative-strand RNA virus obeying the rule of six.