ANTI-TOXIN VACCINE COMPOSITIONS AND METHODS OF USE THEREOF

• US 20120251542A1
• Filed: 03/05/2012
• Published: 10/04/2012
• Est. Priority Date: 03/04/2011
• Status: Active Grant

• Current Assignee: Rutgers University
• Original Assignee: Rutgers University
• Inventors: Li, Xiao-Ping, Tumer, Nilgun
• Granted Patent: US 8,895,006 B2
• Time in Patent Office: Days
• US Class Current: 424/139.1
• CPC Class Codes: A61K 2039/522 avirulent or attenuated
• CPC Class Codes: A61K 39/0258 Escherichia
• CPC Class Codes: A61K 39/0283 Shigella
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/25 Shigella (G)
• CPC Class Codes: G01N 2333/25 Shigella (G)
• CPC Class Codes: G01N 2333/415 from plants
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 2800/42 Poisoning, e.g. from bites or stings
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Compositions and methods for treatment of toxin poisoning are disclosed.

Claims

1. A ricin ribosome binding mutant that is altered relative to a wild type ricin A chain toxin of SEQ ID NO:

1 at at least two amino acid residues, selected from the group consisting of i) R189A and R234A;

ii) R193A and R235A;

iii) R193A and R235A and R189A;

iv) R193A and R235A and R234A;

v) R193A and R235A and R189A and R234A; and

vi) R213A and R258A.

2. The ricin mutant of claim 1, which is R189A and R234A.

3. The ricin mutant of claim 1, which is R193A and R235A.

4. The ricin mutant of claim 1, which is R193A and R235A and R189A.

5. The ricin mutant of claim 1, which is R193A and R235A and R234A.

6. The ricin mutant of claim 1, which is R193A and R235A and R189A and R234A.

7. The ricin mutant of claim 1, which is R213A and R258A.

8. A ricin mutant as claimed in claim 1 wherein the mutant is non-cytotoxic to eukaryotic cells.

9. A composition comprising a mutant ricin toxin as claimed in claim 1 and a biologically acceptable carrier.

10. The composition of claim 9, which is a vaccine, and the ricin mutant is present in an amount effective to elicit an immune response to ricin A chain toxin in an animal.

11. The composition of claim 9, which is a vaccine, and wherein the nucleic acid expresses the ricin mutant in an amount effective to elicit an immune response to ricin A chain toxin in an animal.

12. A method of eliciting an immune response to ricin A chain toxin, comprising contacting an animal with at least one ricin mutant as claimed in claim 1, in an amount effective to elicit an immune response to ricin A chain toxin, wherein an immune response to ricin A chain toxin is elicited in the animal.

13. A method of eliciting an immune response to ricin A chain toxin, comprising contacting an animal with the nucleic acid of claim 11, in an amount effective to express the ricin mutant in an amount effective to elicit an immune response to ricin A chain toxin, wherein an immune response to ricin A chain toxin is elicited in the animal.

14. A shigella toxin ribosome binding mutant that is altered relative to a wild type Stx1 chain toxin of SEQ ID NO:

2 at at least two amino acid residues, selected from the group consisting of

i) R172A and R176A and R179A;

ii) R205A and R220A;

iii) R223A and R248A; and

iv) R251A.

15. The Stx1 mutant of claim 14, which is R172A and R176A and R179A.

16. The Stx1 mutant of claim 14, which is R205A and R220A.

17. The Stx1 mutant of claim 14, which is R223A and R248A.

18. The Stx1 mutant of claim 14 which is R251A.

19. A Stx1 mutant as claimed in claim 14 wherein the mutant is non-cytotoxic to eukaryotic cells.

20. A composition comprising a mutant Stx1 toxin as claimed in claim 14 and a biologically acceptable carrier.

21. The composition of claim 20, which is a vaccine, and the Stx1 mutant is present in an amount effective to elicit an immune response to Stx1 chain toxin in an animal.

22. The composition of claim 20, which is a vaccine, and wherein the nucleic acid expresses the Stx1 mutant in an amount effective to elicit an immune response to Stx1 chain toxin in an animal.

23. A method of eliciting an immune response to Stx1 chain toxin, comprising contacting an animal with at least one Stx1 mutant as claimed in claim 14, in an amount effective to elicit an immune response to Stx1 chain toxin, wherein an immune response to Stx1 chain toxin is elicited in the animal.

24. A method of eliciting an immune response to Stx 1 chain toxin, comprising contacting an animal with the nucleic acid of claim 22, in an amount effective to express the Stx1 mutant in an amount effective to elicit an immune response to Stx1 chain toxin, wherein an immune response to Stx1 chain toxin is elicited in the animal.

25. A shigella toxin ribosome binding mutant that is altered relative to a wild type Stx2 chain toxin of SEQ ID NO:

3 at at least two amino acid residues, selected from the group consisting of

i) R172A and R176A and R179A;

ii) R204A and R219A;

iii) R222A and R247A;

iv) R250A.

26. The Stx2 mutant of claim 25, which is R172A and R176A and R179A.

27. The Stx2 mutant of claim 25, which is R204A and R219A.

28. The Stx2 mutant of claim 25, which is R222A and R247A.

29. The Stx2 mutant of claim 25 which is R250A.

30. A Stx2 mutant as claimed in claim 25 wherein the mutant is non-cytotoxic to eukaryotic cells.

31. A composition comprising a mutant Stx2 toxin as claimed in claim 25 and a biologically acceptable carrier.

32. The composition of claim 31, which is a vaccine, and the Stx2 mutant is present in an amount effective to elicit an immune response to Stx2 chain toxin in an animal.

33. The composition of claim 31, which is a vaccine, and wherein the nucleic acid expresses the Stx2 mutant in an amount effective to elicit an immune response to Stx2 chain toxin in an animal.

34. A method of eliciting an immune response to Stx2 chain toxin, comprising contacting an animal with at least one Stx2 mutant as claimed in claim 25, in an amount effective to elicit an immune response to Stx2 chain toxin, wherein an immune response to Stx2 chain toxin is elicited in the animal.

35. A method of eliciting an immune response to Stx2 chain toxin, comprising contacting an animal with the nucleic acid of claim 33, in an amount effective to express the Stx2 mutant in an amount effective to elicit an immune response to Stx2 chain toxin, wherein an immune response to Stx2 chain toxin is elicited in the animal.

36. A method for identifying neutralizing antibodies to ricin comprising

a) providing a panel of anti-ricin antibodies;

b) identifying those antibodies which bind specifically to the ribosome binding sequences shown in Table 1, said antibodies being effective to prevent ricin binding to the ribosome, thereby neutralizing the toxicity of ricin.

37. A method for treating ricin poisoning comprising passively infusing an effective amount of at least one antibody identified by the method of claim 36 into a patient in need thereof