Double auxotrophic
Provided are attenuated strains of M. tuberculosis and M. bovis BCG which are double auxotrophic mutants with genes knocked out in the biosynthetic pathways for arginine and methionine, and compositions and methods of use thereof.
1. An auxotrophic mycobacterium comprising a mycobacterial genome in which (i) argB gene, or a gene encoding a mycobacterial ArgB enzyme, has been fully or partially deleted and (ii) metA gene, or a gene encoding a mycobacterial MetA enzyme, has been fully or partially deleted, wherein the mycobacterium is sterilized in the absence of arginine and methionine.
2. The auxotrophic mycobacterium of claim 1 , which is incapable of synthesizing arginine and methionine.
3. The auxotrophic mycobacterium of claim 1 , wherein the auxotrophic mycobacterium is a Mycobacterium tuberculosis.
4. The auxotrophic mycobacterium of claim 1 , wherein the auxotrophic mycobacterium is a Mycobacterium bovis Bacillus Calmette-Guérin.
5. The auxotrophic mycobacterium of claim 1 , wherein the auxotrophic mycobacterium is an H37Rv Mycobacterium tuberculosis.
6. A composition comprising the auxotrophic mycobacterium of claim 1 , and a carrier.
7. The composition of claim 6 , which is a vaccine.
8. The composition of claim 6 , wherein the composition does not comprise arginine or methionine.
9. The composition of claim 6 , wherein the composition comprises an amount of arginine and an amount of methionine.
10. A method of eliciting an immune response in a subject comprising administering to the subject the composition of claim 6 in an amount effective to elicit an immune response.
11. A method of vaccinating a subject comprising administering to the subject the composition of claim 6 in an amount effective to vaccinate a subject.
12. A method of treating a subject for tuberculosis comprising administering to the subject the composition of claim 6 in an amount effective to treat a subject.
13. The method of claim 10 , wherein the subject has pulmonary tuberculosis.
14. The method of claim 12 , wherein the tuberculosis is pulmonary tuberculosis.
15. The method of claim 10 , wherein the subject has tuberculosis meningitis.
16. The method of claim 12 , wherein the tuberculosis is tuberculosis meningitis.
17. A method of treating a subject for a cancer comprising administering to the subject the composition of claim 6 in an amount effective to treat a subject.
18. The method of claim 17 , wherein the cancer is a genitourinary cancer.
19. The method of claim 17 , wherein the cancer is a non-invasive bladder cancer.
20. The method of claim 19 , wherein the amount of the composition is administered into the bladder of the subject via a catheter.
21. The method of claim 17 , wherein the subject is immunocompromised.
22. The method of claim 10 , wherein the subject is human.