Trypanosoma antigens, vaccine compositions, and related methods

• US 8,778,348 B2
• Filed: 04/28/2008
• Issued: 07/15/2014
• Est. Priority Date: 04/28/2007
• Status: Active Grant

• Current Assignee: iBio, Inc.
• Original Assignee: iBio, Inc.
• Inventors: Knapp, Elisabeth, Yusibov, Vidadi
• Primary Examiner(s): GRASER, JENNIFER E
• Application Number:
  US12/110,877
• Publication Number:
  US 20090324634A1
• Time in Patent Office: 2,269 Days
• Field of Search: None
• US Class Current: 424/184.1
• CPC Class Codes: A61K 2039/552 Veterinary vaccine
• CPC Class Codes: A61K 2039/6075 Viral proteins
• CPC Class Codes: A61K 39/005 Trypanosoma antigens
• CPC Class Codes: C07K 2319/35 containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
• CPC Class Codes: C07K 2319/40 containing a tag for immunodetection, or an epitope for immunisation
• CPC Class Codes: C12N 15/8257 for the production of primary gene products, e.g. pharmaceutical products, interferon
• CPC Class Codes: C12N 15/8258 for the production of oral vaccines (antigens) or immunoglobulins

Abstract

The present invention relates to the intersection of the fields of immunology and protein engineering, and particularly to antigens and vaccines useful in prevention of infection by Trypanosoma protozoa. Provided are recombinant protein antigens, compositions, and methods for the production and use of such antigens and vaccine compositions.

Claims

1. An isolated antigen comprising a component of a Trypanosoma protein fused to alfalfa mosaic virus (AlMV) coat protein;

wherein the Trypanosoma protein component is a fragment of alpha tubulin or a fragment of beta tubulin consisting of one or more amino acid sequences selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

2. The isolated antigen of claim 1, wherein the Trypanosoma protein component is a polytope.

3. The isolated antigen of claim 1, wherein the Trypanosoma protein component consists of at least two sequences selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

4. The isolated antigen of claim 3, wherein the Trypanosoma protein component consists of a portion of alpha tubulin selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11 and a portion of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20,

a plurality of portions of alpha tubulin selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11, or

a plurality of portions of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

5. A composition comprising a pharmaceutically acceptable carrier and a first antigen comprising a component of a Trypanosoma protein fused to AlMV coat protein;

wherein the Trypanosoma protein component is a fragment of alpha tubulin or a fragment of beta tubulin consisting of one or more amino acid sequences selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

6. The composition of claim 5, wherein the Trypanosoma protein component consists of at least two domains selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

7. The composition of claim 6, wherein the Trypanosoma protein component consists of at least one portion of alpha tubulin selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11 and at least one portion of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20,

a plurality of portions of alpha tubulin selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11, or

a plurality of portions of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

8. The composition of claim 5, further comprising a second antigen, wherein the second antigen comprises a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein,

wherein the Trypanosoma protein component of the second antigen is distinct from the first antigen and is selected from the group consisting of a fragment of alpha tubulin having the sequence set forth in SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, or SEQ ID NO;

11 and a fragment of beta tubulin having the sequence set forth in SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, or SEQ ID NO;

20; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

9. The composition of claim 5, wherein the antigen is produced in a plant selected from a transgenic plant and a plant transiently expressing the antigen.

10. The composition of claim 5, wherein the composition comprises antigen which is purified, partially purified, or unpurified from plant cells, a plant, seeds, fruit, or an extract thereof.

11. A composition comprising at least two antigens, each of which comprises a component of a Trypanosoma protein, wherein the Trypanosoma protein components each consist of an amino acid sequence selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20;

wherein at least one of the antigens is fused to AlMV coat protein;

wherein the composition comprises a pharmaceutically acceptable carrier; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

12. A method for inducing an immune response against Trypanosoma infection in a subject comprising administering to a subject an effective amount of an anti-Trypanosoma composition, wherein the administration is sufficient to stimulate production of antigen specific antibodies or stimulate a cellular immune response by the subject;

thereby inducing an immune response;

wherein the composition comprises an antigen comprising a component of a Trypanosoma protein fused to AlMV coat protein; and

wherein the Trypanosoma protein component is a fragment of alpha tubulin or a fragment of beta tubulin consisting of one or more amino acid sequences selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

13. An isolated antigen comprising a component of a Trypanosoma protein fused to a thermostable protein;

wherein the Trypanosoma protein component is a fragment of alpha tubulin or a fragment of beta tubulin consisting of an amino acid sequence selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

14. The isolated antigen of claim 13, wherein the Trypanosoma protein component is a polytope.

15. The isolated antigen of claim 13, wherein the Trypanosoma protein component consists of at least two sequences selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

16. The isolated antigen of claim 15, wherein the Trypanosoma protein component consists of a portion of alpha tubulin selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11 and a portion of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20,

a plurality of portions of alpha tubulin selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11, or

a plurality of portions of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

17. A composition comprising a pharmaceutically acceptable carrier and a first antigen comprising a component of a Trypanosoma protein fused to a thermostable protein;

wherein the Trypanosoma protein component consists of one or more fragments of alpha tubulin or beta tubulin selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

18. The composition of claim 17, wherein the Trypanosoma protein component consists of at least two domains selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

19. The composition of claim 18, wherein the Trypanosoma protein component consists of at least one portion of alpha tubulin selected from the group consisting of SEQ ID NO:

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11 and at least one portion of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20,

a plurality of portions of alpha tubulin selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, and SEQ ID NO;

11, or

a plurality of portions of beta tubulin selected from the group consisting of SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.

20. The composition of claim 17, further comprising a second antigen, wherein the second antigen comprises a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein and a pharmaceutically acceptable carrier;

wherein the Trypanosoma protein component of the second antigen is distinct from the first antigen and is selected from the group consisting of a fragment of alpha tubulin having the sequence set forth in SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, or SEQ ID NO;

11 and a fragment of beta tubulin having the sequence set forth in SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, or SEQ ID NO;

20; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

21. The composition of claim 17, wherein the antigen is produced in a plant selected from a transgenic plant and a plant transiently expressing the antigen.

22. The composition of claim 17, wherein the composition comprises antigen which is purified, partially purified, or unpurified from plant cells, a plant, seeds, fruit, or an extract thereof.

23. A method for inducing an immune response against Trypanosoma infection in a subject comprising administering to a subject an effective amount of an anti-Trypanosoma composition, wherein the administration is sufficient to stimulate production of antigen specific antibodies or stimulate a cellular immune response by the subject;

thereby inducing an immune response;

wherein the composition comprises an antigen comprising a component of a Trypanosoma protein fused to a thermostable protein; and

wherein the Trypanosoma protein component is a fragment of alpha tubulin or a fragment of beta tubulin consisting of an amino acid sequence selected from the group consisting of SEQ ID NO;

4, SEQ ID NO;

5, SEQ ID NO;

6, SEQ ID NO;

7, SEQ ID NO;

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

16, SEQ ID NO;

18, SEQ ID NO;

19, and SEQ ID NO;

20.