Immunostimulating peptides

1. A peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, and wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS) and wherein a C-terminal carboxyl group is modified.

2. The peptide according to claim 1 , wherein the C-terminal carboxyl group is modified by amidation.

3. The peptide according to claim 1 , wherein the net charge of the peptide at neutral pH is increased compared to the net charge of the peptide having the amino acid sequence SEQ ID NO: 1.

4. The peptide according to claim 1 , wherein the amino acid change(s) is/are independently selected from substitution, deletion, and insertion.

5. The peptide according to claim 1 , wherein the sequence variant comprises 1, 2, or 3 amino acid change(s) in positions 15-17 of SEQ ID NO: 1.

6. The peptide according to claim 5 , wherein the amino acid change is deletion of one or more of residues 15-17.

7. The peptide according to claim 1 , wherein the sequence variant comprises an amino acid change in position 4 of SEQ ID NO: 1.

8. The peptide according to claim 7 , wherein the amino acid change of position 4 is a substitution.

9. The peptide according to claim 8 , wherein the amino acid change in position 4 is a substitution with an amino acid residue selected from the group consisting of amino acids that increase the charge at neutral pH of the peptide compared to SEQ ID NO: 1.

10. The peptide according to claim 9 , wherein position 4 in SEQ ID NO: 1 is substituted with a Lysine residue.

11. The peptide according to claim 1 , wherein the sequence variant comprises an amino acid change in position 5 of SEQ ID NO: 1.

12. The peptide according to claim 11 , wherein the amino acid change of position 5 is a substitution.

13. The peptide according to claim 12 , wherein the amino acid change in position 5 is substitution with an amino acid residue selected from the group consisting of amino acids that increases the charge at neutral pH of the peptide compared to SEQ ID NO: 1.

14. The peptide according to claim 13 , wherein position 5 in SEQ ID NO: 1 is substituted with a Lysine residue.

15. The peptide according to claim 1 , which has the formula I

(I)

(SEQ ID NO: 8)

Z 1 Z 2 Z 3 X 1 X 2 Z 4 Z 5 GLSILLNX 3 X 4 X 5

wherein

Z 1 is L or absent,

Z 2 is Q or absent,

Z 3 is N or absent,

Z 4 , is G or absent,

Z 5 is L or absent,

X 1 is R or K or absent,

X 2 is R or K or absent,

X 3 is E or absent,

X 4 is E or absent, and

X 5 is C or absent,

with the proviso that Formula I does not have the amino acid sequence SEQ ID NO: 1.

16. The peptide according to claim 15 , wherein

if Z 1 is present then Z 2 -Z 5 , X 1 , and X 2 are all present;

if Z 2 is present, then Z 3 -Z 5 , X 1 , and X 2 are all present;

if Z 3 is present, then Z 4 and Z 5 , X 1 , and X 2 are all present;

if Z 4 is present, then Z 5 , X 1 , and X 2 are all present; or

if X 1 is present, then X 2 is present.

17. The peptide according to claim 16 , wherein

Z 1 is absent, or

Z 2 is absent, or

Z 3 is absent, or

Z 4 is absent, or

Z 5 is absent, or

X 1 is absent, or

X 2 is absent.

18. The peptide according to claim 17 , wherein

Z 1 is present, or

Z 2 is present, or

Z 3 is present, or

Z 4 is present, or

Z 5 is present, or

X 1 is present, or

X 2 is present.

19. The peptide according to claim 16 , wherein

Z 1 is present, or

Z 2 is present, or

Z 3 is present, or

Z 4 is present, or

Z 5 is present, or

X 1 is present, or

X 2 is present.

20. The peptide according to claim 15 , wherein

X 3 , X 4 , and X 5 are all present, or

only one of X 3 and X 4 is present and X 5 is present, or

only X 5 is present, or

none of X 3 -X 5 is present.

21. The peptide according to claim 1 , which has the amino acid sequence set forth in any one of SEQ ID NOs: 2-7 and 9.

22. A method for preparation of antibodies that specifically binds an immunogen, the method comprising co-administration to an animal of an immunogenically active amount of

a peptide according to claim 1 , and

the immunogen,

so as to effect production of antibodies specific for said immunogen in said animal, and subsequently recovering said antibodies from the animal.

23. A pharmaceutical composition comprising a peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS), and wherein said composition further comprises a pharmaceutically acceptable carrier, diluent or excipient.

24. The pharmaceutical composition according to claim 23 , which in addition comprises an immunogen.

25. A peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, and wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS) and wherein the sequence variant comprises deletion of one or more of residues 15-17.

26. The peptide according to claim 25 , wherein the net charge of the peptide at neutral pH is increased compared to the net charge of the peptide having the amino acid sequence SEQ ID NO: 1.

27. The peptide according to claim 25 , wherein the amino acid change(s) is/are independently selected from substitution, deletion, and insertion.

28. The peptide according to claim 25 , wherein the sequence variant comprises an amino acid change in position 4 of SEQ ID NO: 1.

29. The peptide according to claim 28 , wherein the amino acid change of position 4 is a substitution.

30. The peptide according to claim 29 , wherein the amino acid change in position 4 is substitution with an amino acid residue selected from the group consisting of amino acids that increase the charge at neutral pH of the peptide compared to SEQ ID NO: 1.

31. The peptide according to claim 30 , wherein position 4 in SEQ ID NO: 1 is substituted with a Lysine residue.

32. The peptide according to claim 25 , wherein the sequence variant comprises an amino acid change in position 5 of SEQ ID NO: 1.

33. The peptide according to claim 32 , wherein the amino acid change of position 5 is a substitution.

34. The peptide according to claim 33 , wherein the amino acid change in position 5 is substitution with an amino acid residue selected from the group consisting of amino acids that increase the charge at neutral pH of the peptide compared to SEQ ID NO: 1.

35. The peptide according to claim 34 , wherein position 5 in SEQ ID NO: 1 is substituted with a Lysine residue.

36. A method for preparation of antibodies that specifically bind an immunogen, the method comprising co-administration to an animal an immunogenically active amount of

a peptide according to claim 25 , and

the immunogen,

so as to effect production of antibodies specific for said immunogen in said animal, and subsequently recovering said antibodies from the animal.

37. A peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, and wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS) and wherein the amino acid in position 4 is substituted with an amino acid residue selected from the group of amino acids that increase the charge at neutral pH of the peptide compared to SEQ ID NO: 1.

38. The peptide according to claim 37 , wherein position 4 in SEQ ID NO: 1 is substituted with a Lysine residue.

39. The peptide according to claim 37 , wherein the sequence variant comprises an amino acid change in position 5 of SEQ ID NO: 1.

40. The peptide according to claim 39 , wherein the amino acid change of position 5 is a substitution.

41. The peptide according to claim 40 , wherein the amino acid change in position 5 is substitution with an amino acid residue selected from the group consisting of amino acids that increase the charge at neutral pH of the peptide compared to SEQ ID NO: 1.

42. The peptide according to claim 41 , wherein position 5 in SEQ ID NO: 1 is substituted with a Lysine residue.

43. A method for preparation of antibodies that specifically bind an immunogen, the method comprising co-administration to an animal an immunogenically active amount of

a peptide according to claim 37 , and

the immunogen,

so as to effect production of antibodies specific for said immunogen in said animal, and subsequently recovering said antibodies from the animal.

44. A peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, and wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS)

wherein the peptide has the formula I

(I)

(SEQ ID NO: 8)

Z 1 Z 2 Z 3 X 1 X 2  Z 4  Z 5 GLSILLNX 3 X 4 X 5  

wherein

Z 1 is L or absent,

Z 2 is Q or absent,

Z 3 is N or absent,

Z 4 , is G or absent,

Z 5 is L or absent,

X 1 is R or K or absent,

X 2 is R or K or absent,

X 3 is E or absent,

X 4 is E or absent, and

X 5 is C or absent,

with the proviso that Formula I does not have the amino acid sequence SEQ ID NO: 1,

and wherein

if Z 1 is present then Z 2 -Z 5 , X 1 , and X 2 are all present;

if Z 2 is present, then Z 3 -Z 5 , X 1 , and X 2 are all present;

if Z 3 is present, then Z 4 and Z 5 , X 1 , and X 2 are all present;

if Z 4 is present, then Z 5 , X 1 , and X 2 are all present; and

if X 1 is present, then X 2 is present,

and wherein

Z 1 is present, or

Z 2 is present, or

Z 3 is present, or

Z 4 is present, or

Z 5 is present, or

X 1 is present, or

X 2 is present.

45. The peptide according to claim 6 , which comprises deletion of one or both of residues 15 and 16 or deletion of residue 17.

46. A pharmaceutical composition comprising a peptide selected from the group consisting of

a peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, and wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS) and wherein the sequence variant comprises deletion of one or more of residues 15-17;

a peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, and wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS) and wherein the amino acid in position 4 is substituted with an amino acid residue selected from the group of amino acids that increase the charge at neutral pH of the peptide compared to SEQ ID NO: 1; and

a peptide consisting of or comprising a sequence variant of the amino acid sequence LQNRRGLGLSILLNEEC (SEQ ID NO: 1), wherein the sequence variant comprises at least one amino acid change compared to SEQ ID NO: 1 in any one of residues 1-7 and 15-17 but no amino acid changes in residues 8-14, and wherein the peptide stimulates secretion and/or expression of cytokines when supplied in an effective concentration to THP-1 cells or PBMCs stimulated with lipopolysaccharide (LPS)

wherein the peptide has the formula I

(I)

(SEQ ID NO: 8)

Z 1 Z 2 Z 3 X 1 X 2  Z 4  Z 5 GLSILLNX 3 X 4 X 5  

wherein

Z 1 is L or absent,

Z 2 is Q or absent,

Z 3 is N or absent,

Z 4 , is G or absent,

Z 5 is L or absent,

X 1 is R or K or absent,

X 2 is R or K or absent,

X 3 is E or absent,

X 4 is E or absent, and

X 5 is C or absent,

with the proviso that Formula I does not have the amino acid sequence SEQ ID NO: 1,

and wherein

if Z 1 is present then Z 2 -Z 5 , X 1 , and X 2 are all present;

if Z 2 is present, then Z 3 -Z 5 , X 1 , and X 2 are all present;

if Z 3 is present, then Z 4 and Z 5 , X 1 , and X 2 are all present;

if Z 4 is present, then Z 5 , X 1 , and X 2 are all present; and

if X 1 is present, then X 2 is present,

and wherein

Z 1 is present, or

Z 2 is present, or

Z 3 is present, or

Z 4 is present, or

Z 5 is present, or

X 1 is present, or

X 2 is present.

47. A method for preparation of antibodies that specifically binds an immunogen, the method comprising co-administration to an animal an immunogenically active amount of

a peptide according to claim 44 , and

the immunogen,

so as to effect production of antibodies specific for said immunogen in said animal, and subsequently recovering said antibodies from the animal.