Deacylation of LPS in gram negative bacteria
The current invention provides new Gram negative polypeptides exhibiting lipid A 3-O-deacylase activity and are capable of modifying and/or detoxifying gram negative LPS. The present invention also provides Gram negative bacteria, Gram negative bacterial lipopolysaccharides (LPS) and compositions comprising LPS, which are provided with or treated with a 3-O-deacylase activity according to the invention and which may be used for pharmaceutical and/or veterinary purposes, in particular for the preparation of whole cell or acellular vaccines against pathogenic Gram negatives such as Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica.
1. A Gram negative bacterium comprising an expression vector that comprises a nucleic acid sequence encoding a polypeptide with lipid A 3-O-deacylase activity, wherein the polypeptide has at least 95% amino acid sequence identity with SEQ ID NO: 1, wherein expression of the nucleic acid sequence increases lipid A 3-O-deacylase activity in said bacterium.
2. The bacterium according to claim 1 which is selected from the group consisting of the following species:
(a) Bordetella pertussis , or
(b) Bordetella parapertussis , or
(c) Bordetella bronchiseptica , or
(d) a Neisseria species.
3. The bacterium according to claim 2 which is a member of the Bordetella pertussis species.
4. The bacterium according to claim 2 , wherein the Neisseria species is selected from the group consisting of N. meningitidis, N. gonorrhoeae and N. lactamica.
5. The bacterium according to claim 1 comprising a partially or fully 3-O-deacylated lipopolysaccharide (LPS) species in its outer membrane.
6. The bacterium according to claim 5 wherein the LPS is palmitoylated.
7. The bacterium according to claim 6 which is a member of the Bordetella pertussis species.
8. The bacterium according to claim 1 wherein the nucleic acid sequence encodes a polypeptide the sequence of which is SEQ ID NO:1.
9. The bacterium according to claim 8 which is selected from the group consisting of the following species:
(a) Bordetella pertussis , or
(b) Bordetella parapertussis , or
(c) Bordetella bronchiseptica , or
(d) a Neisseria species.
10. The bacterium according to claim 9 which is a member of the Bordetella pertussis species.
11. The bacterium according to claim 9 , wherein the Neisseria species is selected from the group consisting of N. meningitidis, N. gonorrhoeae and N. lactamica.
12. The bacterium according to claim 9 comprising a partially or fully 3-O-deacylated lipopolysaccharide (LPS) species in its outer membrane.
13. The bacterium according to claim 12 which is a member of the Bordetella pertussis species.
14. The bacterium according to claim 12 wherein the LPS is palmitoylated.
15. The bacterium according to claim 14 which is a member of the Bordetella pertussis species.
16. A whole cell vaccine comprising the bacterium according to claim 1 and a pharmaceutically acceptable excipient or carrier.
17. A whole cell vaccine comprising the bacterium according to claim 2 and a pharmaceutically acceptable excipient or carrier.
18. A whole cell vaccine comprising the bacterium according to claim 3 and a pharmaceutically acceptable excipient or carrier.
19. A whole cell vaccine comprising the bacterium according to claim 4 and a pharmaceutically acceptable excipient or carrier.
20. A whole cell vaccine comprising the bacterium according to claim 5 and a pharmaceutically acceptable excipient or carrier.
21. A whole cell vaccine comprising the bacterium according to claim 6 and a pharmaceutically acceptable excipient or carrier.
22. A whole cell vaccine comprising the bacterium according to claim 7 and a pharmaceutically acceptable excipient or carrier.
23. A whole cell vaccine comprising the bacterium according to claim 8 and a pharmaceutically acceptable excipient or carrier.
24. A whole cell vaccine comprising the bacterium according to claim 9 and a pharmaceutically acceptable excipient or carrier.
25. A whole cell vaccine comprising the bacterium according to claim 10 and a pharmaceutically acceptable excipient or carrier.
26. A whole cell vaccine comprising the bacterium according to claim 11 and a pharmaceutically acceptable excipient or carrier.
27. A whole cell vaccine comprising the bacterium according to claim 12 and a pharmaceutically acceptable excipient or carrier.
28. A whole cell vaccine comprising the bacterium according to claim 13 and a pharmaceutically acceptable excipient or carrier.
29. A whole cell vaccine comprising the bacterium according to claim 14 and a pharmaceutically acceptable excipient or carrier.
30. A whole cell vaccine comprising the bacterium according to claim 15 and a pharmaceutically acceptable excipient or carrier.
31. A method for treating or preventing a Bordetella infection in a subject, comprising, administering to a subject in need thereof bacteria according to claim 2 , wherein the bacteria are one of said Bordetella species.
32. A method for eliciting an immune response against Bordetella bacteria in a subject, comprising administering to the subject the vaccine according to claim 17 , wherein the bacterium is one of said Bordetella species.