Capsular gram-positive bacteria bioconjugate vaccines
The present invention encompasses a novel S. aureus bioconjugate vaccine. More generally, the invention is directed to Gram-positive and other bioconjugate vaccines containing a protein carrier, at least one polysaccharide such as a capsular Gram-positive polysaccharide, and, optionally, an adjuvant or pharmaceutically acceptable carrier. The instant invention also includes methods of producing Gram-positive and other bioconjugate vaccines. An N-glycosylated protein is also provided that contains one or more polysaccharides such as Gram-positive polysaccharides. The invention is additionally directed to engineered prokaryotic organisms comprising nucleotide sequences encoding a glycosyltransferase of a first prokaryotic organism and a glycosyltransferase of a second prokaryotic organism. The invention further includes plasmids and prokaryotic cells transformed with plasmids encoding polysaccharides and enzymes which produce an N-glycosylated protein and/or bioconjugate vaccine. Further, the invention is directed to methods of inducing an immune response in a mammal comprising administering said bioconjugate vaccines.
1. A Gram-negative host prokaryotic organism comprising:
(i) a nucleotide sequence encoding at least one glycosyltransferase from a Gram-positive bacterium, wherein said Gram-positive bacterium is a Staphylococcus aureus strain that expresses serotype 8 capsular polysaccharide;
(ii) a nucleotide sequence encoding at least one glycosyltransferase from a Gram-negative bacterium;
(iii) a nucleotide sequence encoding a carrier protein, wherein said carrier protein is genetically detoxified Pseudomonas aeruginosa Exotoxin, Staphylococcus aureus ( S. aureus ) alpha hemolysin, or S. aureus clumping factor A; and wherein said carrier protein comprises the amino acid sequence D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; and
(iv) a nucleotide sequence encoding an oligosaccharyl transferase.
2. The host prokaryotic organism of claim 1 , wherein said at least one glycosyltransferase from a Gram-negative bacterium is a P. aeruginosa glycosyltransferase.
3. The host prokaryotic organism of claim 1 , wherein said carrier protein is genetically detoxified P. aeruginosa Exotoxin.
4. The host prokaryotic organism of claim 1 , wherein said carrier protein is S. aureus alpha hemolysin.
5. The host prokaryotic organism of claim 1 , wherein said carrier protein is S. aureus clumping factor A.
6. The host prokaryotic organism of claim 1 , wherein said host organism is E. coli.
7. The host prokaryotic organism of claim 3 , wherein said host organism is E. coli.
8. The host prokaryotic organism of claim 4 , wherein said host organism is E. coli.
9. The host prokaryotic organism of claim 5 , wherein said host organism is E. coli.
10. The host prokaryotic organism of claim 1 , wherein said oligosaccharyl transferase is from Campylobacter jejuni.
11. The host prokaryotic organism of claim 6 , wherein said oligosaccharyl transferase is from Campylobacter jejuni.
12. The host prokaryotic organism of claim 7 , wherein said oligosaccharyl transferase is from Campylobacter jejuni.
13. The host prokaryotic organism of claim 8 , wherein said oligosaccharyl transferase is from Campylobacter jejuni.
14. The host prokaryotic organism of claim 9 , wherein said oligosaccharyl transferase is from Campylobacter jejuni.
15. A composition comprising the host prokaryotic organism of claim 1 .
16. A composition comprising the host prokaryotic organism of claim 2 .
17. A composition comprising the host prokaryotic organism of claim 12 .
18. A composition comprising the host prokaryotic organism of claim 13 .
19. A composition comprising the host prokaryotic organism of claim 14 .
20. A method of producing a bioconjugate comprising a S. aureus Serotype 8 strain capsular polysaccharide linked to genetically detoxified P. aeruginosa Exotoxin, comprising producing in the host prokaryotic organism of claim 3 modified capsular polysaccharides on undecaprenol and linking said modified capsular polysaccharides to genetically detoxified P. aeruginosa Exotoxin.
21. A method of producing a bioconjugate comprising a S. aureus Serotype 8 strain capsular polysaccharide linked to S. aureus alpha hemolysin, comprising producing in the host prokaryotic organism of claim 4 modified capsular polysaccharides on undecaprenol and linking said modified capsular polysaccharides to S. aureus alpha hemolysin.
22. A method of producing a bioconjugate comprising a S. aureus Serotype 8 strain capsular polysaccharide linked to S. aureus clumping factor A, comprising producing in the host prokaryotic organism of claim 5 modified capsular polysaccharides on undecaprenol and linking said modified capsular polysaccharides to P. aeruginosa Exotoxin.
23. A S. aureus bioconjugate comprising, a genetically detoxified P. aeruginosa Exotoxin comprising an inserted consensus sequence D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; a S. aureus serotype 8 strain modified capsular polysaccharide linked to said consensus sequence; and optionally, a pharmaceutically acceptable carrier or adjuvant.
24. A S. aureus bioconjugate comprising, a S. aureus alpha hemolysin further comprising an inserted consensus sequence D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; a S. aureus serotype 8 strain modified capsular polysaccharide linked to said consensus sequence; and optionally, a pharmaceutically acceptable carrier or adjuvant.
25. A S. aureus bioconjugate comprising, a S. aureus clumping factor A comprising an inserted consensus sequence D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; a S. aureus serotype 8 strain modified capsular polysaccharide linked to said consensus sequence; and optionally, a pharmaceutically acceptable carrier or adjuvant.