Recombinant intracellular pathogen immunogenic compositions and methods for use
View Patent ↗Immunogenic compositions comprising recombinant attenuated intracellular pathogens that have been transformed to express recombinant immunogenic antigens of the same or other intracellular pathogens are provided. Exemplary immunogenic compositions include, but are not limited to attenuated recombinant Mycobacteria expressing the major extracellular non-fusion proteins of Mycobacteria and/or other intracellular pathogens. Other embodiments are provided wherein the recombinant attenuated intracellular pathogen is auxotrophic.
1. An immunogenic composition comprising:
a recombinant Bacille Calmette-Guerin (BCG) having an extrachromosomal nucleic acid sequence comprising a gene encoding for a Mycobacterial major extracellular protein selected from the group consisting of 23.5 kDa, 30 kDa, 32 kDa and combinations thereof, wherein said Mycobacterial 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 major extracellular proteins wherein said genes encoding for said Mycobacterial major 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 major extracellular proteins wherein said genes encoding for said Mycobacterial major 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 major 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. 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 mammal.
8. 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 mammal.
9. 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 Mycobacterium tuberculosis (Mtb) 30 kDa major extracellular non-fusion protein and an Mtb 23.5 kDa major extracellular non-fusion protein, wherein said Mtb 30 kDa major extracellular non-fusion 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 mammal.
10. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium bovis 30 kDa major extracellular protein under the control of a promoter wherein said 30 kDa major extracellular protein is over expressed and secreted such that an immune response is induced in a mammal.
11. An immunogenic composition comprising:
a recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium leprae 30 kDa major extracellular protein under the control of a promoter wherein said 30 kDa major extracellular protein is over expressed and secreted such that an immune response is induced in a mammal.
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 Mycobacterium tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular protein are over expressed and secreted;
and wherein said genetic construct comprises a gene encoding for said Mtb 23.5 kDa extracellular protein and a gene encoding for said Mtb 30 kDa extracellular protein and wherein said genes are orientated in the same direction relative to each other within said genetic construct.
13. 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 Mycobacterium tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular protein are over expressed and secreted;
and wherein said genetic construct comprises a gene encoding for said Mtb 23.5 kDa extracellular protein and a gene encoding for said Mtb 30 kDa extracellular protein and wherein said genes are orientated in opposite directions relative to each other within said genetic construct.
14. The immunogenic compositions according to any one of claim 7 , 8 , 9 , 10 or 11 wherein said immune response is a protective immune response.
15. An immunogenic composition comprising:
a growth regulatable recombinant Bacille Calmette-Guérin (BCG) having an extrachromosomal nucleic acid sequence comprising a gene encoding for a Mycobacterial major extracellular protein selected from the group consisting of 23.5 kDa, 30 kDa, 32 kDa and combinations thereof, wherein said Mycobacteria major extraceilular protein is over expressed and secreted.
16. The immunogenic composition according to claim 15 wherein said nucleic acid sequence is under the control of a promoter that is not a heat shock promoter or a stress protein promoter.
17. The immunogenic composition according to claim 15 wherein said major extracellular protein is a non-fusion protein.
18. The immunogenic composition according to claim 15 wherein said extrachromosomal nucleic acid sequence comprises a genetic construct having genes encoding for multiple Mycobacterial major extracellular proteins wherein said genes encoding for said Mycobacterial major extracellular proteins are orientated in the same direction relative to each other within the genetic construct.
19. The immunogenic composition according to claim 15 wherein said extrachromosomal nucleic acid sequence comprises a genetic construct having genes encoding for multiple Mycobacterial major extracellular proteins wherein said genes encoding for said Mycobacterial major extracellular proteins are orientated in opposite directions relative to each other within the genetic construct.
20. The immunogenic compositions according to any one of claims 15 - 19 wherein said Mycobacterial major extracellular proteins are from a species of Mycrobacterium selected from the group consisting of Mycobacterium tuberculosis (Mtb), Mycobacterium bovis (MB), and Mycobacterium leprae (ML).
21. 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 protein wherein said 23.5 kDa major extracellular protein is over expressed and secreted such that an immune response is induced in a mammal.
22. An immunogenic composition comprising:
a growth regulatable recombinant BCG having an extrachromosomal nucleic acid sequence comprising a gene encoding for Mycobacterium tuberculosis 30 kDa major extracellular protein wherein said 30 kDa major extracellular protein is over expressed and secreted such that an immune response is induced in a mammal.
23. 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 wherein said 32A kDa major extracellular non-fusion protein is over expressed and secreted such that an immune response is induced in a mammal.
24. 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 Mycobacterium 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.
25. 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 Mycobacterium tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular protein are over expressed and secreted such that an immune response is induced in a mammal.
26. 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 protein under the control of a promoter wherein said 30 kDa major extracellular protein is over expressed and secreted such that an immune response is induced in a mammal.
27. 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 protein under the control of a promoter wherein said 30 kDa major extracellular protein is over expressed and secreted such that an immune response is induced in a mammal.
28. 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 protein under the control of a promoter wherein said Mycobacterium leprae 30 kDa major extracellular protein is over expressed and secreted from said recombinant BCG such that both a humoral and a cellular immune response is induced in a mammal.
29. 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 Mycobacterium tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular 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.
30. 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 Mycobacterium tuberculosis (Mtb) 30 kDa major extracellular protein and an Mtb 23.5 kDa major extracellular protein, wherein said Mtb 30 kDa major extracellular protein and said Mtb 23.5 kDa major extracellular 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.
31. The immunogenic composition according to any one of claims 15 , 21 , 22 or 23 wherein said growth regulatable recombinant BCG is an auxotroph and wherein tryptophan or glutamine is used to regulate growth of said auxotroph.
32. The immunogenic composition according to any one of claims 15 , 21 , 22 , 25 , 26 , 27 or 28 , wherein said immune response is a protective immune response.
33. The immunogenic composition according to any one of claims 7 , 8 , 10 , 11 , 21 , 22 or 23 wherein said mammal is a human.
34. 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 Mycobacterium major extracellular protein.