Compounds and methods for diagnosis and treatment of leishmaniasis
Compounds and methods are provided for diagnosing, preventing, treating and detecting leishmaniasis infection and stimulating immune responses in patients are disclosed. The compounds disclosed are include polypeptides and fusion proteins that contain at least one immunogenic portion of one or more Leishmania antigens, or a variant thereof. Additionally, methods of screening a screening library for tandem repeat proteins that have immunogenic properties are disclosed. Vaccines and pharmaceutical compositions comprising polynucleotides, polypeptides, fusion proteins and variants thereof that may be used for the prevention and therapy of leishmaniasis, as well as for the detection of Leishmaniasis infection are described.
1. A fusion protein comprising at least a first and second tandem repeat unit,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO:2, 11, 13, 14, 26, 27, 32, 33, 35, and 44; and
wherein the second tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, the amino acid sequence of the first tandem repeat unit.
2. The fusion protein of claim 1 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO:2, 14, 26, 27, 32, 33, 35, and 44.
3. The fusion protein of claim 1 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO: 14, 26, 33 and 44.
4. The fusion protein of claim 1 , further comprising a polypeptide comprising at least 8 consecutive amino acids of, and at least 70% homology to, the amino acid sequence selected from the group consisting of: SEQ ID NO: 119-121.
5. A diagnostic kit for detecting T. cruzi infection in a biological sample, comprising:
one or more fusion protein of claim 1 ; and
a detection reagent.
6. The diagnostic kit of claim 5 , wherein the one or more fusion protein further comprises a polypeptide comprising at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of: SEQ ID NO: 119-121.
7. The diagnostic kit of claim 5 , further comprising a solid support, wherein the one or more fusion protein is bound to a solid support.
8. The diagnostic kit of claim 7 wherein the one or more fusion protein is non-covalently bound to a solid support.
9. The diagnostic kit of claim 8 wherein the solid support comprises one of nitrocellulose, latex and a plastic material.
10. A pharmaceutical composition comprising a physiologically acceptable carrier and one or more fusion protein of claim 1 .
11. An immunogenic composition comprising an immunostimulant and one or more fusion protein of claim 1 .
12. An immunogenic composition according to claim 11 further comprising a delivery vehicle.
13. The fusion protein of claim 1 , wherein the first and second tandem repeat unit are identical.
14. The diagnostic kit of claim 5 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO:2, 14, 26, 27, 32, 33, 35, and 44.
15. The diagnostic kit of claim 5 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO: 14, 26, 33 and 44.
16. The diagnostic kit of claim 5 , wherein the first and second tandem repeat unit are identical.
17. The pharmaceutical composition of claim 10 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO:2, 14, 26, 27, 32, 33, 35, and 44.
18. The pharmaceutical composition of claim 10 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO: 14, 26, 33 and 44.
19. The pharmaceutical composition of claim 10 , wherein the first and second tandem repeat unit are identical.
20. The pharmaceutical composition of claim 10 , wherein the one or more fusion protein further comprises a polypeptide comprising at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of: SEQ ID NO: 119-121.
21. The immunogenic composition of claim 11 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO:2, 14, 26, 27, 32, 33, 35, and 44.
22. The immunogenic composition of claim 11 ,
wherein the first tandem repeat unit comprises an amino acid sequence having at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of SEQ ID NO: 14, 26, 33 and 44.
23. The immunogenic composition of claim 11 , wherein the first and second tandem repeat unit are identical.
24. The immunogenic composition of claim 11 , wherein the one or more fusion protein further comprises a polypeptide comprising at least 8 consecutive amino acids of, and at least 70% homology to, an amino acid sequence selected from the group consisting of: SEQ ID NO: 119-121.