FLAGELLIN FUSION PROTEINS AND USE THEREOF TO INDUCE IMMUNE RESPONSES AGAINST PSEUDOMONAS AERUGINOSA

• US 20120064106A1
• Filed: 03/16/2010
• Published: 03/15/2012
• Est. Priority Date: 03/18/2009
• Status: Active Grant

• Current Assignee: Wake Forest University Health Sciences
• Original Assignee: Wake Forest University Health Sciences
• Inventors: Mizel, Steven B., Wozniak, Daniel J., Weimer, Eric T.
• Granted Patent: US 9,096,659 B2
• Time in Patent Office: Days
• US Class Current: 424/192.1
• CPC Class Codes: A61K 2039/55516 Proteins; Peptides
• CPC Class Codes: A61K 2039/572 cytotoxic response
• CPC Class Codes: A61K 2039/575 humoral response
• CPC Class Codes: A61K 2039/6068 Other bacterial proteins, e.g. OMP
• CPC Class Codes: A61K 39/104 Pseudomonadales, e.g. Pseudomonas
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/21 from Pseudomonadaceae (F)
• CPC Class Codes: C07K 16/1214 from Pseudomonadaceae (F)
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/40 containing a tag for immunodetection, or an epitope for immunisation
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/63 Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression

Abstract

The invention provides compositions and fusion proteins comprising a flagellin adjuvant and a Pseudomonas aeruginosa antigen. The invention further provides pharmaceutical formulations and methods for inducing an immune response against P. aeruginosa (e.g., to prevent and/or treat P. aeruginosa infection).

Claims

1. A fusion protein comprising:

(a) a Pseudomonas aeruginosa type A flagellin adjuvant and/or a Pseudomonas aeruginosa type B flagellin adjuvant; and

(b) a Pseudomonas aeruginosa antigen.

2. The fusion protein of claim 1 comprising:

(a) a Pseudomonas aeruginosa type A flagellin adjuvant and/or a Pseudomonas aeruginosa type B flagellin adjuvant; and

(b) a Pseudomonas aeruginosa OprF antigen and/or a Pseudomonas aeruginosa OprI antigen.

3. The fusion protein of claim 1 comprising:

(a) a Pseudomonas aeruginosa type A flagellin adjuvant; and

(b) a Pseudomonas aeruginosa OprI antigen.

4. The fusion protein of claim 1 comprising:

(a) a Pseudomonas aeruginosa type B flagellin adjuvant; and

(b) a Pseudomonas aeruginosa OprI antigen.

5. The fusion protein of claim 1 comprising:

(a) a Pseudomonas aeruginosa type A flagellin adjuvant; and

(b) a Pseudomonas aeruginosa OprF antigen.

6. The fusion protein of claim 1 comprising:

(a) a Pseudomonas aeruginosa type B flagellin adjuvant; and

(b) a Pseudomonas aeruginosa OprF antigen.

7. The fusion protein of claim 1 comprising:

(a) a Pseudomonas aeruginosa type A flagellin adjuvant; and

(b) a Pseudomonas aeruginosa OprF antigen and a Pseudomonas aeruginosa OprI antigen.

8. The fusion protein of claim 1 comprising:

(a) a Pseudomonas aeruginosa type B flagellin adjuvant; and

(b) a Pseudomonas aeruginosa OprF antigen and a Pseudomonas aeruginosa OprI antigen.

9. The fusion protein of claim 7, wherein the fusion protein comprises a fusion peptide comprising the OprF antigen and the OprI antigen.

13. The fusion protein of claim 1, wherein the Pseudomonas aeruginosa antigen(s) is an N-terminal extension of the flagellin adjuvant.

14. The fusion protein of claim 1, wherein the Pseudomonas aeruginosa antigen(s) is between the N-terminal constant region and the C-terminal constant region of the flagellin adjuvant.

15. The fusion protein of claim 14, wherein the flagellin adjuvant(s) comprises a deleted hypervariable region or the hypervariable region is absent from the flagellin adjuvant(s).

16. The fusion protein of claim 14, wherein the Pseudomonas aeruginosa antigen(s) is inserted (i) within the hypervariable region, (ii) between the N-terminal constant region and the hypervariable region, or (iii) between the C-terminal constant region and the hypervariable region.

17. A nucleic acid encoding the fusion protein of claim 1.

18. A vector comprising the nucleic acid of claim 17.

19. A host cell comprising the nucleic acid of claim 17.

20. A method of making a fusion protein of, the method comprising culturing the host cell of claim 19 in a culture medium under conditions sufficient for the fusion protein to be produced.

21. An immunogenic formulation comprising the fusion protein of claim 1 in a pharmaceutically acceptable carrier.

22. A method of producing an immune response against Pseudomonas aeruginosa in a mammalian subject, the method comprising administering the fusion protein of claim 1 to the mammalian subject in an amount effective to produce an immune response in the mammalian subject against Pseudomonas aeruginosa.

23. A method of protecting a mammalian subject from infection with Pseudomonas aeruginosa, the method comprising administering the fusion protein of claim 1 to the mammalian subject in an amount effective to protect the mammalian subject from infection with Pseudomonas aeruginosa.

24. A method of enhancing a protective immune response to Pseudomonas aeruginosa in a mammalian subject, the method comprising administering the fusion protein of claim 1 to the mammalian subject in an amount effective to enhance the protective immune response to Pseudomonas aeruginosa in the mammalian subject

25. A method of treating a mammalian subject infected with motile, nonmucoid Pseudomonas aeruginosa, the method comprising administering the fusion protein of claim 1 to the mammalian subject in an amount effective to treat the mammalian subject infected with motile, nonmucoid Pseudomonas aeruginosa.

26. The method of claim 22, wherein the administering step is carried out by intramuscular delivery, intradermal delivery, subcutaneous delivery, or by delivery to a mucosal surface.

27. The method of any of claim 22, wherein the mammalian subject is a human subject.

28. The method of claim 27, wherein the human subject is afflicted with cystic fibrosis, is afflicted with a burn injury, is immunocompromised and/or is on a ventilator.

29. The method of claim 28, wherein the human subject is afflicted with cystic fibrosis and administration occurs within two years of diagnosis.

30. The method of claim 28, wherein the human subject is afflicted with cystic fibrosis and administration occurs within two years of a first infection with P. aeruginosa.

10. A composition comprising:

(a) a fusion protein comprising (i) a Pseudomonas aeruginosa type A flagellin adjuvant; and

(ii) a Pseudomonas aeruginosa antigen; and

(b) a fusion protein comprising (i) a Pseudomonas aeruginosa type B flagellin adjuvant; and

(ii) a Pseudomonas aeruginosa antigen.

11. The composition of claim 10 comprising:

(a) a fusion protein comprising (i) a Pseudomonas aeruginosa type A flagellin adjuvant; and

(ii) a Pseudomonas aeruginosa OprF antigen and/or a Pseudomonas aeruginosa OprI antigen; and

(b) a fusion protein comprising (i) a Pseudomonas aeruginosa type B flagellin adjuvant; and

(ii) a Pseudomonas aeruginosa OprF antigen and/or a Pseudomonas aeruginosa OprI antigen.

12. The composition of claim 10 comprising:

(a) a fusion protein comprising (i) a Pseudomonas aeruginosa type A flagellin adjuvant; and

(ii) a Pseudomonas aeruginosa OprF antigen and a Pseudomonas aeruginosa OprI antigen; and

(b) a fusion protein comprising (i) a Pseudomonas aeruginosa type B flagellin adjuvant; and

(ii) a Pseudomonas aeruginosa OprF antigen and a Pseudomonas aeruginosa OprI antigen.