Mycobacterial mutants affecting host apoptosis
View Patent ↗Provided are recombinant mycobacteria having a mutation in an nlaA gene or in a nuoG gene. Also provided are isolated and purified nlaA proteins and nuoG proteins from a mycobacterium . Additionally provided are isolated and purified nucleic acids comprising a recombinant nlaA gene or a recombinant nuoG gene. Further provided are methods of inducing an immune response in a mammal and methods of making a recombinant mycobacterium using the nlaA gene or the nuoG gene.
1. A recombinant mycobacterium having deletion of nlaA gene or deletion of a portion of nlaA gene, wherein the mutation deletion increases the ability of the mycobacterium to induce apoptosis of a mammalian macrophage infected by the recombinant mycobacterium.
2. The mycobacterium of claim 1 , wherein the nlaA gene encodes a protein that is at least 99% homologous to SEQ ID NO: 1.
3. The mycobacterium of claim 1 , deletion is of the entire nlaA gene (ΔnlaA).
4. The mycobacterium of claim 1 , wherein the deletion is of a portion of the nlaA gene.
5. The mycobacterium of claim 1 , further comprising a mutation, wherein the mycobacterium exhibits attenuated virulence in a mammal when compared to the mycobacterium without the mutation.
6. The mycobacterium of claim 5 , wherein the mutation is a deletion in at least a portion of an RD1 region, or a deletion in a gene controlling production of a vitamin or an amino acid.
7. The mycobacterium of claim 1 , wherein the mycobacterium is M. smegmatis, M. bovis, M. avium, M. phlei, M. fortuitum, M. lufu, M. paratuberculosis, M. habana, M. scrofulacium, M. intracellulare, M. tuberculosis , or M. kansasii.
8. The mycobacterium of claim 7 , wherein the mycobacterium is an M. boris.
9. The mycobacterium of claim 1 , wherein the mycobacterium is M. tuberculosis.
10. The mycobacterium of claim 1 , further comprising a recombinant gene operably linked to a promoter that directs expression of the gene when the mycobacterium infects a mammalian cell.
11. The mycobacterium of claim 10 , wherein the gene encodes an antigen of a mammalian pathogen.
12. A method of inducing an immune response in a mammal, the method comprising inoculating the mammal with the mycobacterium of claim 1 .
13. The method of claim 12 , wherein the mycobacterium further comprises a mutation in a nuoG gene, wherein the mutation increases the ability of the mycobacterium to induce apoptosis of a mammalian macrophage infected by the mycobacterium.
14. A method of making a recombinant mycobacterium , the method comprising eliminating expression of the nlaA gene in the mycobacterium.