Claims
- 1. An immunogenic composition comprising: recombinant Bacille Calmette-Guerin (BCG) having an extrachromosomal nucleic acid sequence comprising a gene encoding for a Mycobacteria tuberculosis major extracellular protein selected from the group consisting of 23.5 kDa, 32 kDa and combinations thereof, wherein said Mycobacteria tuberculosis major extracellular protein is over expressed and secreted
- 2. The immunogenic composition according to claim 1 wherein said nucleic acid sequence is under the control of a promoter that is not a heat shock promoter or a stress protein promoter.
- 3. The immunogenic composition according to claim 1 wherein said major extracellular protein is a non-fusion protein.
- 4. The immunogenic composition according to claim 1 wherein said extrachromosomal nucleic acid sequence comprises a genetic construct having genes encoding for multiple Mycobacterial extracellular proteins wherein said genes encoding for said Mycobacterial extracellular proteins are orientated in the same direction relative to each other within the genetic construct.
- 5. The immunogenic composition according to claim 1 wherein said extrachromosomal nucleic acid sequence comprises a genetic construct having genes encoding for multiple Mycobacterial extracellular proteins wherein said genes encoding for said Mycobacterial extracellular proteins are orientated in opposite directions relative to each other within the genetic construct.
- 6. The immunogenic compositions according to any one of claims 1-5 wherein said Mycobacterial extracellular proteins are from a species of Mycobacterium selected from the group consisting of Mycobacterium tuberculosis (Mtb), Mycobacterium bovis (MB), and Mycobacterium leprae (ML).
- 7. The immunogenic compositions according to claim 6 wherein said Mycobacterial extracellular protein is selcted from the group consisting of Mtb 23.5 kDa protein, Mtb 30 kDa protein, Mtb 32A kDa protein, MB 30 kDa protein, MB 32A kDa protein, ML 23.5 kDa protein, ML 30 kDa protein and ML 32A kDa protein.
- 8. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium tuberculosis 23.5 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 23.5 kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a host.
- 9. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium tuberculosis 23.5 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium tuberculosis 23.5 kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 10. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium tuberculosis 32A kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 32A kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a host.
- 11. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid comprising a gene encoding for Mycobacterium tuberculosis 32A kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium tuberculosis 32A kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 12. An immunogenic composition comprising:
a recombinant Bacille Calmette-Guérin (BCG) having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted.
- 13. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted such that an immune response is induced in a host.
- 14. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium bovis 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 30 kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a host.
- 15. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid comprising a gene encoding for Mycobacterium bovis 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium bovis 30 kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 16. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium leprae 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 30 kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in an animal.
- 17. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid comprising a gene encoding for Mycobacterium leprae 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium leprae 30 kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 18. An immunogenic composition comprising:
a recombinant Bacille Calmette-Guérin (BCG) having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted; and wherein said genetic construct comprises a gene encoding for said Mtb 23.5 kDa protein and a gene encoding for said Mtb 30 kDa protein and wherein said genes are orientated in the same direction relative to each other within said genetic construct.
- 19. An immunogenic composition comprising:
a recombinant Bacille Calmette-Guérin (BCG) having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted; and wherein said genetic construct comprises a gene encoding for said Mtb 23.5 kDa protein and a gene encoding for said Mtb 30 kDa protein and wherein said genes are orientated in opposite directions relative to each other within said genetic construct.
- 20. An immunogenic composition comprising: a growth regulatable recombinant Bacille Calmette-Guerin (BCG) having an extrachromosomal nucleic acid sequence comprising a gene encoding for a Mycobacteria tuberculosis major extracellular protein selected from the group consisting of 23.5 kDa, 32 kDa and combinations thereof, wherein said Mycobacteria tuberculosis major extracellular protein is over expressed and secreted
- 21. The immunogenic composition according to claim 20 wherein said nucleic acid sequence is under the control of a promoter that is not a heat shock promoter or a stress protein promoter.
- 22. The immunogenic composition according to claim 20 wherein said major extracellular protein is a non-fusion protein.
- 23. The immunogenic composition according to claim 20 wherein said extrachromosomal nucleic acid sequence comprises a genetic construct having genes encoding for multiple Mycobacterial extracellular proteins wherein said genes encoding for said Mycobacterial extracellular proteins are orientated in the same direction relative to each other within the genetic construct.
- 24. The immunogenic composition according to claim 20 wherein said extrachromosomal nucleic acid sequence comprises a genetic construct having genes encoding for multiple Mycobacterial extracellular proteins wherein said genes encoding for said Mycobacterial extracellular proteins are orientated in opposite directions relative to each other within the genetic construct.
- 25. The immunogenic compositions according to any one of claims 20-24 wherein said Mycobacterial extracellular proteins are from a species of Mycobacterium selected from the group consisting of Mycobacterium tuberculosis (Mtb), Mycobacterium bovis (MB), and Mycobacterium leprae (ML).
- 26. The immunogenic compositions according to claim 25 wherein said Mycobacterial extracellular protein is selected from the group consisting of Mtb 23.5 kDa protein, Mtb 30 kDa protein, Mtb 32A kDa protein, MB 30 kDa protein, MB 32A kDa protein, ML 23.5 kDa protein, ML 30 kDa protein and ML 32A kDa protein.
- 27. An immunogenic composition comprising:
a growth reguatable recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium tuberculosis 23.5 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 23.5 kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a host.
- 28. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium tuberculosis 23.5 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium tuberculosis 23.5 kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 29. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium tuberculosis 32A kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 32A kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a host.
- 30. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid comprising a gene encoding for Mycobacterium tuberculosis 32A kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium tuberculosis 32A kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 31. An immunogenic composition comprising:
a growth regulatable recombinant Bacille Calmette-Guérin (BCG) having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted.
- 32. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted such that an immune response is induced in a host.
- 33. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium bovis 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 30 kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a host.
- 34. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid comprising a gene encoding for Mycobacterium bovis 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium bovis 30 kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 35. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium leprae 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said 30 kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a host.
- 36. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid comprising a gene encoding for Mycobacterium leprae 30 kDa major extracellular non-fusion protein under the control of a promoter wherein said promoter is not a heat shock promoter or stress protein promoter and wherein said Mycobacterium leprae 30 kDa major extracellular non-fusion protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a host.
- 37. An immunogenic composition comprising:
a growth regulatable recombinant Bacille Calmette-Guérin (BCG) having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted; and wherein said genetic construct comprises a gene encoding for said Mtb 23.5 kDa protein and a gene encoding for said Mtb 30 kDa protein and wherein said genes are orientated in the same direction relative to each other within said genetic construct.
- 38. An immunogenic composition comprising:
a growth regulatable recombinant Bacille Calmette-Guérin (BCG) having an extrachromosomal nucleic acid sequence comprising a genetic construct having at least one gene encoding for a Mycobacteria tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular non-fusion protein are over expressed and secreted; and wherein said genetic construct comprises a gene encoding for said MTB 23.5 kDa protein and a gene encoding for said Mtb 30 kDa protein and wherein said genes are orientated in opposite directions relative to each other within said genetic construct.
- 39. The immunogenic composition according to any one of claims 20 or 27 to 30 wherein said growth regulatable recombinant BCG is an auxotroph and wherein tryptophan or glutamine is used to regulate growth of said auxotroph.
RELATED APPLICATIONS
[0001] This application is a continuation-in-part of application Ser. No. 10/261,981 filed Sep. 30, 2002 which is a continuation-in-part of application Ser. No. 09/550,468 filed Apr. 17, 2000, now U.S. Pat. No. 6,471,967, the entire contents of which are hereby incorporated by reference in their entirety.
REFERENCE TO GOVERNMENT
[0002] This invention was made with Government support under Grant No. AI31338 awarded by the Department of Health and Human Services. The Government has certain rights in this invention.
Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
10261981 |
Sep 2002 |
US |
Child |
10439611 |
May 2003 |
US |
Parent |
09550468 |
Apr 2000 |
US |
Child |
10261981 |
Sep 2002 |
US |