Recombinant biologically contained filovirus vaccine
AssigneeWisconsin Alumni Research Foundation (WARF)
InventorsYoshihiro Kawaoka, Peter J. Halfmann
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Abstract
The invention provides a vaccine comprising a recombinant biologically contained filovirus and methods of making and using those viruses.
Claims18
A vaccine comprising an effective amount of a recombinant filovirus and one or more adjuvants comprising a saponin, wherein the genome of the recombinant filovirus contains a deletion of one or more nucleotides in a polynucleotide sequence for a viral protein corresponding to Ebola virus VP30, and wherein the deletion is effective to prevent expression of a functional viral protein corresponding to Ebola virus VP30 upon infection of a cell with the recombinant filovirus, wherein the genome further comprises a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product and wherein the nucleotide sequence is inserted into the filovirus genome at a site other than the site of the deletion in the polynucleotide or wherein the genome further comprises a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product and the nucleotide sequence replaces or is inserted into GP/sGP sequences or a portion thereof.
A multivalent vaccine comprising an effective amount of a recombinant filovirus and comprising a saponin, wherein the genome of the recombinant filovirus contains a first deletion in one or more nucleotides for a polynucleotide sequence for a viral protein corresponding to Ebola virus VP30 which deletion is effective to prevent expression of a functional viral protein corresponding to Ebola virus VP30 upon infection of a cell with the recombinant filovirus, wherein the genome further comprises a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product and wherein the nucleotide sequence is inserted into the filovirus genome at a site other than the site of the deletion in the polynucleotide or wherein the genome further comprises a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product and the nucleotide sequence replaces or is inserted into GP/sGP sequences or a portion thereof, and wherein the genome encodes one or more filovirus glycoproteins.
A method of manufacturing a composition comprising recombinant filovirus and an adjuvant, comprising:
- providing supernatant from mammalian cells expressing a recombinant filovirus genome that are cultured in serum free medium so as to result in progeny filovirus, wherein the genome of the recombinant filovirus contains a deletion of one or more nucleotides in a polynucleotide sequence for a viral protein corresponding to Ebola virus VP30 which deletion is effective to prevent expression of a functional viral protein corresponding to Ebola virus VP30 upon infection of a cell with the recombinant filovirus, wherein the genome further comprises a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product, and wherein the nucleotide sequence is inserted into the filovirus genome at a site other than the site of the deletion in the polynucleotide or wherein the nucleotide sequence replaces or is inserted into GP/sGP sequences or a portion thereof, and wherein the mammalian cells express a viral protein corresponding to Ebola virus VP30;
- contacting the supernatant with a DNase and a virus inactivating agent, thereby providing an inactivated viral preparation;
- concentrating the inactivated viral preparation;
- and
- combining the inactivated viral preparation and one or more saponin adjuvants.
A vaccine comprising an effective amount of a recombinant filovirus and one or more adjuvants comprising a squalene or a saponin, wherein the genome of the recombinant filovirus contains a deletion of one or more nucleotides in a polynucleotide sequence for a viral protein corresponding to Ebola virus VP30, and wherein the deletion is effective to prevent expression of a functional viral protein corresponding to Ebola virus VP30 upon infection of a cell with the recombinant filovirus, wherein the genome further comprises a nucleotide sequence encoding a prophylactic or therapeutic heterologous gene product, and wherein the nucleotide sequence is inserted into the filovirus genome at a site other than the site of the deletion in the polynucleotide or wherein the nucleotide sequence replaces or is inserted into GP/sGP sequences or a portion thereof.
Dependencies are inferred from the claim text.