Immunostimulatory recombinant intracellular pathogen immunogenic compositions and methods of use
View Patent ↗Immunogenic compositions comprising recombinant intracellular pathogens that have been transformed to express recombinant immunogenic antigens of the same or other intracellular pathogens and immunostimulatory molecules are provided. Exemplary immunogenic compositions include, but are not limited to, recombinant BCG expressing Mycobacteria major extracellular proteins and immunostimulatory molecules.
1. An immunogenic composition for inducing a protective immune response to Mycobacterium tuberculosis in a host, the composition comprising:
a recombinant Bacille Calmette Guérin (rBCG) expressing:
a 30 kDa M. tuberculosis major extracellular protein, wherein said 30 kDa M. tuberculosis major extracellular protein is over-expressed and secreted; and
interferon gamma, wherein said interferon gamma is expressed from an extrachromosomal nucleic acid,
wherein said 30 kDa M. tuberculosis major extracellular protein and said interferon gamma are expressed under the control of a promoter that is not a heat shock promoter.
2. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein is expressed on an extrachromosomal nucleic acid sequence.
3. The immunogenic composition according to claim 1 wherein said interferon gamma is expressed on an extrachromosomal nucleic acid sequence.
4. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein and said interferon gamma are expressed from different extrachromosomal nucleic acid sequences.
5. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein and said interferon gamma are expressed from the same extrachromosomal nucleic acid sequence.
6. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein is integrated into the rBCG genome under the control of a promoter that is not a heat shock promoter and over-expressed.
7. The immunogenic composition according to claim 1 wherein said interferon gamma is integrated into the rBCG genome under the control of a promoter that is not a heat shock promoter and expressed.
8. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein and said interferon gamma are integrated into the rBCG genome under the control of a promoter that is not a heat shock promoter and over-expressed.
9. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein is integrated into the rBCG genome under the control of a promoter that is not a heat shock promoter and over-expressed and said interferon gamma is expressed on an extrachromosomal nucleic acid sequence.
10. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein is a non-fusion protein.
11. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein is a fusion protein.
12. The immunogenic composition according to claim 1 wherein said M. tuberculosis major extracellular protein is over-expressed and secreted such that a protective immune response is induced in a host.
13. The immunogenic composition according to claim 1 further expressing at least one additional Mycobacteria major extracellular protein selected from the group consisting of 12 kDa protein, 14 kDa protein, 16 kDa protein, 23.5 kDa protein, 24 kDa protein, 32A kDa protein, 32B kDa protein, 45 kDa protein, 58 kDa protein, 71 kDa protein, 80 kDa protein, and 110 KD protein.
14. The immunogenic composition according to claim 13 wherein said at least one additional Mycobacteria major extracellular protein is from a species of Mycobacterium selected from the group consisting of Mycobacterium tuberculosis; Mycobacterium bovis, mycobacterium leprae , and Mycobacterium avium intracellulare.
15. The immunogenic composition of according to claim 1 wherein said interferon gamma is a covalent dimer comprising one copy of an interferon gamma gene linked to a 19 amino acid peptide linker which is in turn linked to a second copy of an interferon gamma gene.
16. The immunogenic composition according to claim 1 wherein said interferon gamma is expressed in the absence of a signal peptide.
17. The immunogenic composition of claim 1 wherein said rBCG is growth-regulatable.
18. The immunogenic composition of claim 17 wherein said rBCG is siderophore-deficient.
19. A method of inducing a protective immune response to M. tuberculosis in a host comprising:
administering the immunogenic composition of claim 1 to said host, wherein a protective immune response to M. tuberculosis is induced in said host.
20. The method of claim 19 wherein said host has previously been immunized with a BCG-containing vaccine.