Recombinant BCG strains with enhanced ability to escape the endosome
Mycobacterium strains that have an enhanced ability to elicit a MHC-Class I-restricted CD8 + T cell immune response are provided. The Mycobacterium strains are genetically engineered to express: a endosomolytic protein that is active at neutral pH (e.g. Perfringolysin O), permitting escape of the Mycobacterium from endosomes into the cytoplasm of the cell; and antigens of interest, such as tuberculosis antigens. The invention also provides vaccine preparations containing such Mycobacterium strains.
1. A recombinant Mycobacterium that is genetically engineered to include a nucleic acid sequence that encodes an expressable and secretable Prefringolysin O (PFO) protein that is active at a pH of 6-8.
2. The recombinant Mycobacterium of claim 1 , further comprising nucleic acid sequences that encode one or more proteins of interest.
3. The recombinant Mycobacterium of claim 2 , wherein said one or more proteins of interest include one or more Plasmodium antigens.
4. The recombinant Mycobacterium of claim 1 , wherein said recombinant Mycobacterium is an attenuated Mycobacterium.
5. The recombinant Mycobacterium of claim 4 , wherein said attenuated Mycobacterium is Bacille Calmette Guerin (BCG).
6. The recombinant Mycobacterium of claim 5 , wherein said BCG is BCG Danish 1331.
7. The recombinant Mycobacterium of claim 1 , wherein said nucleic acid sequence that encodes an expressable and secretable PFO protein is present in a chromosome of said recombinant Mycobacterium.
8. The recombinant Mycobacterium of claim 1 , wherein said nucleic acid sequence that encodes an expressable and secretable PFO protein replace a urease C gene in a chromosome of said recombinant Mycobacterium.
9. The recombinant Mycobacterium of claim 2 , wherein said nucleic acid sequences that encode one or more proteins of interest are present in a chromosome of said recombinant Mycobacterium.
10. The recombinant Mycobacterium of claim 2 , wherein said one or more proteins of interest of interest include Mycobacterium tuberculosis (Mtb) antigens.
11. The recombinant Mycobacterium of claim 10 , wherein said Mtb antigens are selected from the group consisting of Ag85A, Ag85B, TB 10.4, Rv0125, Rv0203, Rv0287, Rv0288, Rv0603, Rv1196, Rv1223, Rv1271c, Rv1733c, Rv1738 Rv1804c, Rv1886, Rv2031c, Rv2032, Rv2253, Rv2290, Rv2389c, Rv2626c, Rv2627c, Rv2779c, Rv2873, Rv2875, Rv3017c, Rv3407, Rv3804c, Rv3810, and Rv3841.
12. The recombinant Mycobacterium of claim 2 , wherein said nucleic acid sequences that encode one or more proteins of interest are present on a plasmid.
13. A composition comprising a recombinant Mycobacterium that is genetically engineered to include: a nucleic acid sequence that encodes an expressable and secretable Prefringolysin O (PFO) protein that is active at a pH of 6-8; and nucleic acid sequences that encode one or more proteins of interest.
14. The composition of claim 13 , wherein said one or more proteins of interest of interest include Mycobacterium tuberculosis (Mtb) antigens.
15. The composition of claim 14 , wherein said Mtb antigens are selected from the group consisting of Ag85A, Ag85B, TB 10.4, Rv0125, Rv0203, Rv0287, Rv0288, Rv0603, Rv1196, Rv1223, Rv1271c, Rv1733c, Rv1738 Rv1804c, Rv1886, Rv2031c, Rv2032, Rv2253, Rv2290, Rv2389c, Rv2626c, Rv2627c, Rv2779c, Rv2873, Rv2875, Rv3017c, Rv3407, Rv3804c, Rv3810, and Rv3841.
16. The composition of claim 13 , wherein said nucleic acid sequences that encode one or more proteins of interest are present on a plasmid.
17. The composition of claim 13 , wherein said recombinant Mycobacterium is an attenuated Mycobacterium.
18. The composition of claim 17 , wherein said attenuated Mycobacterium is Bacille Calmette Guerin (BCG).
19. The composition of claim 18 , wherein said BCG is BCG Danish 1331.
20. The composition of claim 13 , wherein said nucleic acid sequence that encodes an expressable and secretable PFO protein is present in a chromosome of said recombinant Mycobacterium.
21. The composition of claim 13 , wherein said nucleic acid sequence that encodes an expressable and secretable PFO protein replace a urease C gene in a chromosome of said recombinant Mycobacterium.
22. The composition of claim 13 , wherein said nucleic acid sequences that encode one or more proteins of interest are present in a chromosome of said recombinant Mycobacterium.
23. The composition of claim 18 , wherein said composition is a vaccine.
24. The composition of claim 23 , wherein said vaccine is a vaccine for Mycobacterium tuberculosis.
25. The composition of claim 13 , wherein said composition further comprises an adjuvant.
26. The composition of claim 13 , wherein said composition further comprises a pharmacologically suitable carrier.
27. The composition of claim 13 , wherein said one or more proteins or interest include Plasmodium antigens.