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OwnerManinder Sidhu, John Eldridge +2inventors
InventorsManinder Sidhu, John Eldridge, Michael Egan, Zimra Israel
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The invention provides a DNA plasmid comprising: (a) a first transcriptional unit comprising a nucleotide sequence that encodes a first polypeptide operably linked to regulatory elements including a first promoter and a first polyadenylation signal; (b) a second transcriptional unit comprising a nucleotide sequence that encodes a second polypeptide operably linked to regulatory elements including a second promoter and a second polyadenylation signal; (c) a third transcriptional unit comprising a nucleotide sequence that encodes a third polypeptide operably linked to regulatory elements including a third promoter and a third polyadenylation signal; and wherein said first, said second and said third promoters are each derived from different transcriptional units; and wherein said first, said second and said third polyadenylation signals are each derived from different transcriptional units. The invention further relates to immunogenic compositions for inducing an immune response to HIV comprising combinations of two, three, or four plasmids, where each plasmid is expressing a defined antigen, which may be a single antigen or a fusion of two or three antigens.
A DNA plasmid comprising:
2-3. (canceled)
6-25. (canceled)
27-29. (canceled)
An immunogenic composition for inducing an immune response to selected antigens in a vertebrate host, said immunogenic composition comprising:
32-36. (canceled)
A method of immunizing a vertebrate host against selected antigens comprising administering to said vertebrate host an immunogenic composition comprising:
The method of claim 7, wherein said immunogenic composition is administered to a mammal using in vivo electroporation.
The method of claim 8, wherein said electroporation involves electrically stimulating the muscle with an electrical current having a field strength in the range of from about 25 V/cm to about 800 V/cm.
40-42. (canceled)
An immunogenic composition for inducing an immune response to human immunodeficiency virus (HIV) in a vertebrate host, said immunogenic composition comprising:
The immunogenic composition of claim 10, wherein said transfection facilitating agent is bupivacaine.
The immunogenic composition of claim 10, wherein said promoters are selected from the group consisting of human cytomegalovirus (HCMV) immediate early promoter, the simian cytomegalovirus (SCMV) promoter, the murine cytomegalovirus (MCMV) promoter, the herpes simplex virus (HSV) latency-associated promoter-1 (LAP1), Simian virus 40 promoter, human elongation factor 1 alpha promoter, and the human muscle cell specific desmin promoter.
The immunogenic composition of claim 10, wherein said polyadenylation signals are selected from the group consisting of rabbit beta-globin poly(A) signal, synthetic polyA, HSV Thymidine kinase poly A, Human alpha globin poly A, SV40 poly A, human beta globin poly A, polyomavirus poly A, and Bovine growth hormone poly A.
The immunogenic composition of claim 12, wherein said promoter on said first plasmid is the human cytomegalovirus (HCMV) immediate early promoter.
The immunogenic composition of claim 13, wherein said polyadenylation signal on said first plasmid is the Bovine growth hormone poly A polyadenylation signal.
The immunogenic composition of claim 10, wherein said first DNA plasmid encodes an HIV gag-pol fusion polypeptide, wherein said fusion of the gag and pol genes of HIV or gag-pol gene is derived from the HXB2 isolate of HIV.
The immunogenic composition of claim 12, wherein said first promoter on said second plasmid is the human cytomegalovirus (HCMV) immediate early promoter.
The immunogenic composition of claim 13, wherein said first polyadenylation signal on said second plasmid is the SV40 poly A polyadenylation signal.
The immunogenic composition of claim 10, wherein said HIV nef-tat-vif fusion polypeptide is a nef, tat, and vif (NTV) fusion protein expressed from a fusion of the nef, tat, and vif (ntv) genes of HIV.
The immunogenic composition of claim 19, wherein said fusion of the nef, tat, and vif genes of HIV or ntv gene is derived from the NL4-3 isolate of HIV.
The immunogenic composition of claim 12, wherein said second promoter on said second plasmid is the simian cytomegalovirus (SCMV) promoter.
The immunogenic composition of claim 13, wherein said second polyadenylation signal on said second plasmid is the Bovine growth hormone (BGH) polyadenylation signal.
The immunogenic composition of claim 10, wherein said HIV envelope polypeptide is derived from the primary isolate 6101 of HIV.
A method of immunizing a vertebrate host against selected antigens comprising administering to said vertebrate host an immunogenic composition comprising:
The method of claim 24, wherein said immunogenic composition is administered to a mammal using in vivo electroporation.
The method of claim 25, wherein said electroporation involves electrically stimulating the muscle with an electrical current having a field strength in the range of from about 25 V/cm to about 800 V/cm.
The method of claim 24, wherein said transfection facilitating agent is bupivacaine.
The method of claim 24, wherein said promoters are selected from the group consisting of human cytomegalovirus (HCMV) immediate early promoter, the simian cytomegalovirus (SCMV) promoter, the murine cytomegalovirus (MCMV) promoter, the herpes simplex virus (HSV) latency-associated promoter-1 (LAP1), Simian virus 40 promoter, human elongation factor 1 alpha promoter, and the human muscle cell specific desmin promoter.
The method of claim 24, wherein said polyadenylation signals are selected from the group consisting of rabbit beta-globin poly(A) signal, synthetic polyA, HSV Thymidine kinase poly A, Human alpha-globin poly A, SV40 poly A, human beta globin poly A, polyomavirus poly A, and Bovine growth hormone poly A.
The method of claim 28, wherein said promoter on said first plasmid is the human cytomegalovirus (HCMV) immediate early promoter.
The method of claim 29, wherein said polyadenylation signal on said first plasmid is the Bovine growth hormone poly A polyadenylation signal.
The method of claim 24, wherein said first DNA plasmid encodes an HIV gag-pol fusion polypeptide, wherein said fusion of the gag and pol genes of HIV or gag-pol gene is derived from the HXB2 isolate of HIV.
The method of claim 28, wherein said first promoter on said second plasmid is the human cytomegalovirus (HCMV) immediate early promoter.
The method of claim 29, wherein said first polyadenylation signal on said second plasmid is the SV40 poly A polyadenylation signal.
The method of claim 24, wherein said HIV nef-tat-vif fusion polypeptide is a nef, tat, and vif (NTV) fusion protein expressed from a fusion of the nef, tat, and vif (ntv) genes of HIV.
The method of claim 35, wherein said fusion of the nef, tat, and vif genes of HIV or ntv gene is derived from the NL4-3 isolate of HIV.
The method of claim 28, wherein said second promoter on said second plasmid is the simian cytomegalovirus (SCMV) promoter.
The method of claim 29, wherein said second polyadenylation signal is the Bovine growth hormone (BGH) polyadenylation signal.
The method of claim 24, wherein said HIV envelope polypeptide is derived from the primary isolate 6101 of HIV.
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Dependencies are inferred from the claim text.