Recombinant mycobacterium avium subsp. paratuberculosis proteins induce immunity and protect against infection

Patent metadata

Field	Value
Publication number	US-9175049-B2
Application number	US-201314038077-A
Country	US
Kind code	B2
Filing date	Sep 26, 2013
Priority date	Oct 24, 2012
Publication date	Nov 3, 2015
Grant date	Nov 3, 2015

Abstract

Compositions of immunogenic proteins of Mycobacterium avium subspecies paratuberculosis (MAP) are effective for stimulating a protective immune response in animals against MAP. Combinations of MAP1087 with two or more of MAP1204, MAP1272c and MAP2077c are effective for stimulating a protective immune response in animals against MAP and can be administered as vaccines against paratuberculosis (Johne's disease). Induction of the immune response significantly reduces or eliminates colonization of the animal by MAP, and consequently reduces or eliminates the symptoms of clinical disease in animals infected with MAP and reduces or eliminates fecal shedding of MAP. Vaccination with the compositions provides protection against clinical disease and reduces transmission of MAP infection within a herd.

First claim

We claim:

1. A composition comprising isolated and purified immunogenic proteins of Mycobacterium avium subspecies paratuberculosis and a pharmaceutically acceptable carrier, wherein said immunogenic proteins are provided in an amount effective to stimulate an immune response in an animal against said Mycobacterium avium subspecies paratuberculosis , and said immunogenic proteins comprise MAP1087 and at least two of MAP1204, MAP1272c, and MAP2077c wherein said MAP1087 comprises SEQ ID NO: 1; wherein said MAP1204 comprises SEQ ID NO: 2; wherein said MAP1272c comprises SEQ ID NO: 3; and wherein said MAP2077c comprises SEQ ID NO: 4, wherein said pharmaceutically acceptable carrier is selected from the group consisting of physiological saline, phosphate buffered saline, mineral oil, vegetable oil, aqueous carboxymethyl cellulose, and polyvinylpyrrolidone.

2. The composition of claim 1 further comprising an adjuvant.

3. The composition of claim 1 wherein said immunogenic proteins comprise MAP1087, MAP1204 and MAP1272c.

4. The composition of claim 1 wherein said immunogenic proteins comprise MAP1087, MAP1204 and MAP2077c.

5. The composition of claim 1 wherein said immunogenic proteins comprise MAP1087, MAP1272c and MAP2077c.

6. The composition of claim 1 wherein said immunogenic proteins comprise MAP1087, MAP1204, MAP1272c and MAP2077c.

7. An immunogenic composition comprising fusion protein MAP1087/MBP and at least two of the fusion proteins MAP1204/MBP, MAP1272c/MBP, and MAP2077c/MBP wherein said MAP1087/MBP comprises SEQ ID NO: 5, said MAP1204/MBP comprises SEQ ID NO: 6, said MAP1272c/MBP comprises SEQ ID NO: 7 and said MAP2077c/MBP comprises SEQ ID NO: 8.

8. A method for reducing Mycobacterium avium subspecies paratuberculosis colonization in an animal comprising administering to said animal an immunologically effective dose of a composition comprising a pharmaceutically acceptable carrier, isolated and purified MAP1087 and at least two of isolated and purified MAP1204, isolated and purified MAP1272c, and isolated and purified MAP2077c; wherein said MAP1087 comprises SEQ ID NO: 1, MAP1204 comprises SEQ ID NO: 2, MAP1272c comprises SEQ ID NO: 3, and MAP2077c comprises SEQ ID NO: 4, wherein said immunologically effective dose causes said animal to generate an immune response sufficient to protect said animal from infection with said Mycobacterium avium subspecies paratuberculosis ; and wherein said pharmaceutically acceptable carrier is selected from the group consisting of physiological saline, phosphate buffered saline, mineral oil, vegetable oil, aqueous carboxymethyl cellulose, and polyvinylpyrrolidone.

9. The method of claim 8 wherein said animal is a ruminant.

10. The method of claim 9 wherein said animal is selected from the group consisting of bovine, ovine and caprine.

11. The method of claim 8 wherein said composition further comprises an adjuvant.

12. The method of claim 8 wherein said composition comprises MAP1087, MAP1204 and MAP1272c.

13. The method of claim 8 wherein said composition comprises MAP1087, MAP1204 and MAP2077c.

14. The method of claim 8 wherein said composition comprises MAP1087 and MAP1272c and MAP2077c.

15. The method of claim 8 wherein said composition comprises MAP1087, MAP1204, MAP1272c and MAP2077c.

16. A method for reducing Mycobacterium avium subspecies paratuberculosis colonization in an animal comprising administering to said animal an immunologically effective dose of a immunogenic composition comprising fusion protein MAP1087/MBP and at least two of the fusion proteins MAP1204/MBP, MAP1272c/MBP, and MAP2077c/MBP, wherein said MAP1087/MBP comprises SEQ ID NO: 5, said MAP1204/MBP comprises SEQ ID NO: 6, said MAP1272c/MBP comprises SEQ ID NO: 7 and said MAP2077c/MBP comprises SEQ ID NO: 8, wherein said immunologically effective dose causes said animal to generate an immune response sufficient to protect said animal from infection with said Mycobacterium avium subspecies paratuberculosis.

17. The method of claim 16 wherein said animal is a ruminant.

18. The method of claim 16 wherein said composition further comprises an adjuvant.

19. A method of generating a Th1 immune response against Mycobacterium avium subspecies paratuberculosis in an animal comprising administering to said animal a composition in an amount sufficient to generate a Th1 immune response in said animal, said composition comprising a pharmaceutically acceptable carrier, isolated and purified MAP1087 and at least two of isolated and purified MAP1204, isolated and purified MAP1272c, and isolated and purified MAP2077c; wherein said MAP1087 comprises SEQ ID NO: 1, MAP1204 comprises SEQ ID NO: 2, MAP1272c comprises SEQ ID NO: 3, and MAP2077c comprises SEQ ID NO: 4; and wherein said pharmaceutically acceptable carrier is selected from the group consisting of physiological saline, phosphate buffered saline, mineral oil, vegetable oil, aqueous carboxymethyl cellulose, and polyvinylpyrrolidone.

20. The method of claim 19 wherein said animal is a ruminant.

21. The method of claim 19 wherein said composition further comprises an adjuvant.

22. The method of claim 19 wherein said composition comprises MAP1087, MAP1204, and MAP1272c.

23. The method of claim 19 wherein said composition comprises MAP1087, MAP1204, and MAP2077c.

24. The method of claim 19 wherein said composition comprises MAP1087, MAP1272c, and MAP2077c.

25. The method of claim 19 wherein said composition comprises MAP1087, MAP1204, MAP1272c, and MAP2077c.

26. A method of generating a Th1 immune response against Mycobacterium avium subspecies paratuberculosis in an animal comprising administering to said animal an composition in an amount sufficient to generate a Th1 immune response in said animal, said composition comprising fusion protein MAP1087/MBP and at least two of the fusion proteins MAP1204/MBP, MAP1272c/MBP, and MAP2077c/MBP, wherein said MAP1087/MBP comprises SEQ ID NO: 5, said MAP1204/MBP comprises SEQ ID NO: 6, said MAP1272c/MBP comprises SEQ ID NO: 7 and said MAP2077c/MBP comprises SEQ ID NO: 8.

27. The method of claim 26 wherein said animal is a ruminant.

28. The method of claim 26 wherein said composition further comprises an adjuvant.

29. The method of claim 28 wherein said amount contains approximately 0.1 mg to approximately 0.5 mg of said MAP1087/MBP, and approximately 0.1 mg to approximately 0.5 mg of each of said at least two of MAP1204/MBP, MAP1272c/MBP, and MAP2077c/MBP.

30. The method of claim 26 wherein said amount contains approximately 0.1 mg to approximately 0.5 mg of said MAP1087/MBP, and approximately 0.1 mg to approximately 0.5 mg of each of said at least two of MAP1204/MBP, MAP1272c/MBP, and MAP2077c/MBP.

31. The method of claim 19 wherein said amount contains approximately 0.1 mg to approximately 0.5 mg of said MAP1087/MBP, and approximately 0.1 mg to approximately 0.5 mg of each of said at least two of MAP1204/MBP, MAP1272c/MBP, and MAP2077c/MBP.

32. The method of claim 21 wherein said amount contains approximately 0.1 mg to approximately 0.5 mg of said MAP1087/MBP, and approximately 0.1 mg to approximately 0.5 mg of each of said at least two of MAP1204/MBP, MAP1272c/MBP, and MAP2077c/MBP.

33. The method of claim 16 wherein said immunolog

Assignees

• Us Agriculture

Inventors

• Stabel Judith R
• Bannantine John P

Classifications

• C07K14/35 Primary

  from Mycobacteriaceae (F) · CPC title

• A61P37/00

  Drugs for immunological or allergic disorders · CPC title

• A61K2039/552

  Veterinary vaccine · CPC title

• A61K2039/6068

  Other bacterial proteins, e.g. OMP · CPC title

• A61K39/04 Primary

  Mycobacterium, e.g. Mycobacterium tuberculosis · CPC title