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AssigneeSt. Louis University
InventorsRobert B. Belshe, Ranjit Ray
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The present invention is based upon the observation that the temperature sensitive phenotype of the cp45 strain of HPIV-3 correlates to a mutation in the large, or L, gene of cp45 relative to the corresponding gene in the wild-type strain. This correlation enables new vaccines directed at viruses other than HPIV-3 by combining, through genetic engineering methods, the region of the cp45 viral genome which encodes proteins responsible for replication and internal structure with the region of the genome of the target virus which encodes proteins responsible for attachment, penetration and release of the virus and virus progeny, respectfully. Moreover, it is possible to determine whether HPIV-3 or a cp45-hybrid virus is attenuated by confirming the presence or absence of mutations in its L gene.
A hybrid virus comprising
A live, attenuated vaccine comprising
A live, attenuated human vaccine comprising
A hybrid virus comprising an enveloped, negative-sense, single-stranded chimeric RNA genome, the genome including, in succession from its 3' end:
A hybrid virus comprising an enveloped, negative-sense, single-stranded chimeric RNA genome, the genome including, in succession from its 3' end:
A method for producing an enveloped, negative-sense, single-stranded RNA virus, the method comprising:
A plasmid vector comprising a chimeric RNA genome which includes (i) a nucleic acid sequence which encodes each surface antigen of an enveloped, negative-sense, single-strand RNA target virus selected from the group consisting of HPIV-1, HPIV-2 and RSV, and (ii) a nucleic acid sequence which encodes a HPIV-3 large protein, L, the variant L protein having at least two substitutions in amino acid sequence relative to the L protein of wild-type HPIV-3 (JS), the substitutions being His for Tyr at residue 942 of SEQ ID NO:1 and Phe for Leu at residue 992 of SEQ ID NO:1, and having an RNA polymerase activity which is at least about 10% less than the polymerase activity of the target virus at a temperature of about 39.degree. C., the sense of the genome being either positive or negative.
A method for determining whether an enveloped, negative-sense, single-stranded RNA virus having a genome which encodes a HPIV-3 large protein, L is attenuated, the method comprising confirming the presence of at least one variation in the genome of the virus relative to the genome of wild-type HPIV-3 (JS), the variations being in the region of the genome which encodes the L protein and effecting at least one substitution in the L-Protein of the virus relative to the L-protein of wild-type HPIV-3(JS), the substitution being selected from the group consisting of His for Tyr at residue 942 of SEQ ID NO:1, and Phe for Leu at residue 992 of SEQ ID NO:1 and Ile for Thr at residue 1558 of SEQ ID NO:1.
A method for determining whether an enveloped, negative-sense, single-stranded RNA virus having a genome which encodes a HPIV-3 large protein, L, is attenuated, the method comprising
Dependencies are inferred from the claim text.