Recombinant intracellular pathogen immunogenic compositions and methods of 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 growth regulatable recombinant Bacille Calmette-Guerin (rBCG) having a first extrachromosomal nucleic acid sequence comprising a gene encoding for a first Mycobacteria major extracellular protein selected from the group consisting of 30 kDa protein, 23.5 kDa protein, 32A kDa protein and combinations thereof;
wherein said Mycobacteria major extracellular proteins are over expressed and secreted and said growth regulatable rBCG is selected from the group consisting of auxotrophs and metabolically impaired mutants and combinations thereof.
2. The immunogenic composition according to claim 1 further comprising a second extrachromosomal nucleic acid sequence comprising a gene encoding for a second Mycobacteria major extracellular protein selected from the group consisting of 30 kDa, protein 23.5 kDa protein, 32A kDa protein and combinations thereof.
3. The immunogenic composition according to claim 2 wherein at least one of said first extrachromosomal nucleic acid sequence and second extrachromosomal nucleic acid sequence is under the control of a promoter that is not a heat shock promoter or a stress protein promoter.
4. The immunogenic composition according to claim 2 wherein at least one of said major extracellular proteins are non-fusion proteins.
5. The immunogenic composition according to claim 2 wherein said first or said second Mycobacteria major extracellular protein is from a species of Mycobacterium selected from the group consisting of Mycobacterium tuberculosis (Mtb), Mycobacterium bovis (MB), and Mycobacterium leprae (ML).
6. The immunogenic composition according to claim 4 wherein said extracellular non-fusion proteins are over expressed and secreted such that a protective immune response is induced in a host.
7. The immunogenic composition according to claim 1 wherein said metabolically impaired mutant is a siderophore mutant.
8. The immunogenic composition according to claim 7 wherein said siderophore is a mycobactin or an exochelin.
9. The immunogenic composition according to claim 1 wherein said growth regulatable rBCG is an auxotroph and wherein tryptophan, glutamine or pantothenic acid is used to regulate growth of said auxotroph.