Immunogenic compositions against tuberculosis
Methods of preparing mutants of Mycobacterium tuberculosis with one or more disrupted genes are presented, where the disrupted genes include ctpV, rv0990c, rv0971c, and/or rv0348. Compositions containing mutants with attenuated virulence and pathogenesis, which are capable of stimulation of an immune response against tuberculosis, are described. Compositions and methods relating to immunogenic compositions, which include an attenuated M. tb strain in which the M. tb genome includes a disruption of at least one of the ctpV gene, the rv0990c gene, the rv0971c gene, and the rv0348 gene, are also provided.
1. An engineered Mycobacterium tuberculosis strain whose genome comprises a disruption of at least one gene selected from the group consisting of a ctpV gene, an rv0990c gene and an rv0971c gene.
2. The engineered Mycobacterium tuberculosis strain of claim 1 , wherein mice infected with the engineered M. tuberculosis strain have an increased average post-infection lifespan of at least 125% compared to mice infected with the corresponding wild-type strain.
3. The engineered Mycobacterium tuberculosis strain of claim 1 , wherein the genome comprises a disruption of the rv0990c gene.
4. The engineered Mycobacterium tuberculosis strain of claim 1 , wherein the genome comprises a disruption of the rv0971c gene.
5. The engineered Mycobacterium tuberculosis strain of claim 1 , wherein the genome comprises a disruption of the ctpV gene.
6. The engineered Mycobacterium tuberculosis strain of claim 5 , wherein the engineered M. tuberculosis strain has an average lifespan in 500 μM CuCl 2 , which is decreased by at least 25% as compared to a corresponding wild-type M. tuberculosis strain.
7. An immunogenic composition comprising: an attenuated Mycobacterium tuberculosis strain having a genome comprising a disruption of at least one gene selected from the group consisting of a ctpV gene, an rv0990c gene and an rv0971c gene.
8. The immunogenic composition of claim 7 , further comprising a pharmaceutically acceptable carrier.
9. The immunogenic composition of claim 7 , further comprising a pharmaceutically acceptable adjuvant.
10. A method of stimulating an immune response against tuberculosis, comprising: administering to a subject an immunogenic composition according to claim 7 .
11. An engineered attenuated Mycobacterium tuberculosis strain whose genome comprises a disruption of at least one gene selected from the group consisting of a ctpV gene, an rv0990c gene and an rv0971c gene.
12. The engineered attenuated Mycobacterium tuberculosis strain of claim 11 , wherein the disruption prohibits transcription of a full-length wild-type mRNA from the disrupted gene.
13. The engineered attenuated Mycobacterium tuberculosis strain of claim 11 , wherein one or more disrupted genes comprise an insertion of a gene cassette.
14. The engineered attenuated Mycobacterium tuberculosis strain of claim 11 , wherein one or more disrupted genes comprise replacement of at least a portion of the gene with a gene cassette.
15. The engineered attenuated Mycobacterium tuberculosis strain as in claim 13 , wherein the gene cassette includes a nucleotide sequence encoding a selectable marker.
16. The engineered attenuated Mycobacterium tuberculosis strain of claim 11 , wherein one or more disrupted gene exhibits decreased expression of the respective disrupted gene product.