Recombitope peptides
View Patent ↗The present invention provides peptides having T cell stimulating activity termed recombitope peptides. Recombitope peptides of the invention preferably comprise at least two T cell epitopes derived from the same or from different protein antigens, and more preferably comprise at least two regions, each region preferably having human T cell stimulating activity and each region comprising at least one T cell epitope derived from a protein antigen. Recombitope peptides of the invention can be derived from protein allergens, autoantigens, or other protein antigens. The invention also provides methods of diagnosing sensitivity to a protein allergen or other protein antigen in an individual, methods to treat such sensitivity and therapeutic compositions comprising one or more recombitope peptides. The invention further provides methods for designing recombitope peptides of the invention where the protein antigen to which the individual is sensitive has unknown or ill-defined T cell epitopes.
1. A method for designing an isolated peptide having an amino terminus and a carboxy terminus comprising at least two regions each having human T cell stimulating activity, wherein said regions comprise an epitope of a protein allergen selected from one or both of Dermatophagoides pteronyssinus I (Der p I) and Dermatophagoides pteronyssinus II (Der p II), and wherein said regions are arranged from the amino terminus to the carboxy terminus in a different order than the regions in the naturally-occurring protein allergen, comprising the steps of:
a) dividing amino acid sequence of the protein allergen into at least two regions;
b) determining whether the regions of step (a) have T cell stimulating activity;
c) arranging the regions to form at least one peptide in which the regions are arranged from the amino terminus to the carboxy terminus in a different order than the regions in the naturally-occurring protein allergen; and
d) producing at least one peptide having the arrangement of the regions of step (c).
2. The method of claim 1 , further comprising the step of determining whether the peptide of step (d) has T cell stimulating activity.
3. The method of claim 1 , further comprising the step of determining whether the peptide binds immunoglobulin E specific for the protein allergen of step (a).
4. The method of claim 1 , wherein in step (a) the protein allergen is divided into overlapping regions.
5. The method of claim 1 , wherein at least one T cell epitope of the protein allergen is known and utilized as at least one of said regions.
6. The method of claim 1 , wherein step (b) further comprises determining whether said regions bind immunoglobulin E specific for the allergen of step (a) and cause the release of mediators from mast cells or basophils.
7. The method of claim 1 , wherein the peptide is produced recombinantly.
8. The method of claim 1 , wherein the peptide is produced synthetically.
9. The method of claim 1 , wherein the peptide further comprises a proteolytic site inserted between at least two of said regions.
10. The method of claim 1 , wherein each region comprises at least two T cell epitopes of the protein allergen.
11. The method of claim 1 , wherein each region comprises at least about thirty amino acid residues of the protein allergen.
12. The method of claim 1 , wherein the peptide comprises at least about forty amino acid residues of the protein allergen.
13. The method of claim 1 , wherein the peptide comprises at least about fifteen per cent of the T cell epitopes of the protein allergen.
14. The method of claim 1 , wherein the peptide comprises at least about thirty per cent of the T cell epitopes of the protein allergen.
15. A method for designing an isolated peptide having an amino terminus and a carboxy terminus comprising at least two regions each having human T cell stimulating activity, wherein each region comprises an amino acid sequence selected from the group consisting of sequence X (SEQ ID NO: 7), sequence Y (SEQ ID NO: 8), sequence Z (SEQ ID NO: 9), sequence A (SEQ ID NO: 10), and sequence B (SEQ ID NO: 11), and wherein said regions are arranged from the amino terminus to the carboxy terminus in a different order than the regions in the naturally-occurring protein allergen, comprising the steps of:
a) dividing the protein allergen into at least two regions;
b) determining whether the regions of step (a) have T cell stimulating activity;
c) arranging the regions to form at least one peptide in which the regions are arranged from the amino terminus to the carboxy terminus in a different order than the regions in the naturally-occurring protein allergen; and
d) producing at least one peptide having the arrangement of the regions of step (c).