SALMONELLA VACCINE MATERIALS AND METHODS

• US 20040009191A1
• Filed: 03/15/2001
• Published: 01/15/2004
• Est. Priority Date: 03/17/2000
• Status: Active Grant

• Current Assignee: Pharmacia & Upjohn Company (Pfizer Inc.)
• Original Assignee: Pharmacia & Upjohn Company (Pfizer Inc.)
• Inventors: Lowery, David E., Kennedy, Michael J.
• Granted Patent: US 6,923,957 B2
• Time in Patent Office: Days
• US Class Current: 424/200.1
• CPC Class Codes: A61K 2039/522 avirulent or attenuated
• CPC Class Codes: A61K 2039/523 expressing foreign proteins
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/12 Antidiarrhoeals
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 14/255 Salmonella (G)

Abstract

Attenuated mutant Salmonella bacteria containing inactivated virulence genes are provided for use in safe, efficacious vaccines.

Claims

1. A vaccine composition comprising an immunologically protective amount of a first attenuated, non-reverting mutant Salmonella bacterium in which two or more genes within the SPI2 region have been inactivated.

2. The vaccine composition of claim 1 wherein said genes are inactivated by deletion of a portion of the coding region of the gene.

3. The vaccine composition of claim 1 wherein said genes are inactivated by an insertional mutation.

4. The vaccine composition of claim 1 wherein the genes are (ssa) secretion system apparatus genes.

5. The vaccine composition of claim 1 wherein the genes are selected from the group consisting of ssaT, ssaJ, ssaC and ssaM, and wherein:

(a) said ssaT gene consists of SEQ ID NO;

1 or 2, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO;

1 or 2 under stringent conditions, or a full length Salmonella nucleotide sequence that has 95% sequence identity to SEQ ID NO;

1 or 2;

(b) said ssaJ gene consists of SEQ ID NO;

3 or 4, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO;

3 or 4 under stringent conditions, or a full length Salmonella nucleotide sequence that has 95% sequence identity to SEQ ID NO;

3 or 4;

(c) said ssaC gene consists of SEQ ID NO;

5 or 6, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO;

5 or 6 under stringent conditions, or a full length Salmonella nucleotide sequence that has 95% sequence identity to SEQ ID NO;

5 or 6; and

(d) said ssaM gene consists of SEQ ID NO;

7 or 30, or a full length nucleotide sequence that hybridizes to the non coding complement of SEQ ID NO;

7 or 30 under stringent conditions, or a full length Salmonella nucleotide sequence that has 95% sequence identity to SEQ ID NO;

7 or 30.

6. The vaccine composition of claim 5 wherein, in said first attenuated mutant Salmonella bacterium, ssaT and ssaC have been inactivated.

7. The vaccine composition of claim 2 wherein, in said first attenuated mutant Salmonella bacterium, ssaT and ssaJ have been inactivated.

8. The vaccine composition of claim 1 further comprising a second attenuated mutant Salmonella bacterium in which one or more genes within the SPI2 region have been inactivated.

9. The vaccine composition of claim 8 wherein said genes are selected from the group consisting of ssaT, ssaJ, ssaC, and ssaM.

10. The vaccine composition of claim 8 wherein said first and second mutant Salmonella bacteria are from different serogroups.

11. The vaccine composition of claim 1 or 8 wherein said Salmonella bacteria are Salmonella enterica subsp Enterica.

12. The vaccine composition of claim 1 or 8 wherein said Salmonella bacteria are from any of serogroups A, B, C₁, C₂, D, or E₁.

13. The vaccine composition of claim 8 wherein said first and second attenuated mutant Salmonella bacteria are selected from the group consisting of S. dublin and S. typhimurium.

14. The vaccine composition of claim 1 wherein said first attenuated mutant Salmonella bacterium further comprises a polynucleotide encoding a non-Salmonella polypeptide.

28. A method of delivering a polypeptide antigen to an animal comprising the step of administering the vaccine composition of claim 13 to said animal.

15. A method of conferring protective immunity on a non-rodent animal comprising the step of administering to said animal a vaccine composition comprising an immunologically protective amount of an attenuated, non-reverting mutant Salmonella bacterium in which one or more genes within the SPI2 region have been inactivated.

16. The method of claim 15 wherein said immunologically protective amount of said attenuated bacterium provides an improvement in mortality, symptomatic diarrhea, physical condition, or milk production.

17. The method of claim 15 wherein said gene is inactivated by deletion of a portion of the coding region of the gene.

18. The method of claim 15 wherein said gene is a secretion system apparatus (ssa) gene.

19. The method of claim 15 wherein said genes are selected from the group consisting of ssaT, ssaJ, ssaC and ssaM.

25. The method of claim 15 or 20 wherein said animal is selected from the group consisting of cattle, sheep, goats, horses, pigs, poultry and other birds, cats, dogs, and humans.

26. The method of claim 15 or 20 wherein said animal is a pig.

27. The method of claim 15 or 20 wherein said animal is cattle.

20. A method of reducing the amount or duration of bacterial shedding during infection comprising the step of administering to a non-rodent animal a vaccine composition comprising an attenuated, non-reverting mutant Salmonella bacterium in which one or more genes within the SPI2 region have been inactivated, in an amount effective to reduce bacterial shedding by said animal.

21. The method of claim 20 wherein the amount of bacterial shedding is reduced by about 10% or more.

22. The method of claim 20 wherein the duration of bacterial shedding is reduced by about 10% or more.

23. The method of claim 20 wherein said gene is a secretion system apparatus (ssa) gene.

24. The methods of claim 20 wherein said gene is selected from the group consisting of ssaT, ssaJ, ssaC, and ssaM.