Immunogenic peptides of the cyclin b1 tumor antigen

Abstract

The present invention relates to peptides comprising at least one CD4 + T epitope, which is immunodominant in vitro, of the cyclin B1 tumor antigen, said peptides being capable of stimulating a specific human CD4 + T lymphocyte response in subjects who have varied HLA II molecules, and to the use of these peptides as a cancer vaccine and as a reagent for the diagnosis of cancer or the immunomonitoring of the cellular response against cyclin B1 during cancer or during an anticancer treatment.

1 : An isolated peptide, consisting of a sequence of from 11 to 30 amino acids which is derived from the human cyclin B1 sequence of SEQ ID No. 1 and which comprises at least one human cyclin B1 CD4 +  T epitope which is immunodominant in vitro, said peptide being selected from the group consisting of:
 a) the sequences of 11 to 30 consecutive amino acids of the human cyclin B1 sequence of SEQ ID No. 1 comprising at least residues 3 to 11, 19 to 27, 376 to 384, 27 to 35, 120 to 128, 282 to 290, 287 to 295, 305 to 313, 317 to 325, 322 to 330, 325 to 333, 379 to 387, 412 to 420, 418 to 426 or 422 to 430 of said sequence SEQ ID No. 1, and 
 b) the sequences of 11 to 30 amino acids having at least 70% identity with a sequence in a), 
 
       with the exclusion of the sequence of 15 amino acids made up of residues 279 to 293 of said sequence SEQ ID No. 1, 
       said peptide in a) and b) being capable of stimulating a specific human CD4 +  T lymphocyte response. 
     
     
         2 : The isolated peptide as claimed in  claim 1 , characterized in that the average strength of the in vitro response of human CD4 +  T lymphocytes specific for said peptide is at least 2.5% in a group of human individuals expressing varied HLA II molecules including at least the HLA-DR1, HLA-DR3, HLA-DR4, HLA-DR7, HLA-DR11, HLA-DR13 and HLA-DR15 molecules, and the frequency of responders in vitro to said peptide is at least 55% in said group of human individuals. 
     
     
         3 : The isolated peptide as claimed in  claim 1 , selected from the group consisting of the sequences SEQ ID Nos. 10, 12, 13, 26, 57, 58, 61, 63, 64, 65, 76, 77, 82, 83, 84, 87, 88, 89 and 91. 
     
     
         4 : A mixture of peptides comprising: (i) at least one first peptide comprising a cyclin B1 CD4 +  T epitope which is immunodominant in vitro, as claimed in  claim 1 , and (ii) at least one second peptide comprising at least one CD4 +  T epitope other than said epitope which is immunodominant in vitro, and/or one CD8 +  T epitope, and/or one B epitope. 
     
     
         5 : The mixture of peptides as claimed in  claim 4 , wherein said second peptide is a peptide comprising a cyclin B1 CD4 +  T epitope, capable of binding to at least 6 different predominant HLA II molecules chosen from the HLA-DR1, HLA-DR3, HLA-DR4, HLA-DR7, HLA-DR11, HLA-DR13, HLA-DR15, HLA-DRB3, HLA-DRB4, HLA-DRB5, HLA-DP401 and HLA-DP402 molecules, selected from the group made up of: a) the sequences of 11 to 30 consecutive amino acids of the human cyclin B1 sequence of SEQ ID No. 1 comprising at least residues 170-178, 201-209, 204-212, 209-217, 214-222, 218-226, 223-231, 227-235, 243-251, 246-254, 252-260, 269-277, 301-309, 344-352, 365-373, 368-376 or 371-379 of said sequence SEQ ID No. 1, and b) the sequences of 11 to 30 amino acids having at least 70% identity with a sequence in a). 
     
     
         6 : The mixture of peptides as claimed in  claim 5 , wherein said second peptide is selected from the group consisting of the sequences SEQ ID Nos. 33, 39, 40, 41, 42, 43, 44, 45, 48, 49, 50, 54, 60, 69, 73, 74, 75, 85, 86 and 90. 
     
     
         7 : The mixture of peptides as claimed in  claim 4 , wherein said second peptide is a peptide comprising a cyclin B1 CD4 +  T epitope, capable of binding to 3 to 5 different predominant HLA II molecules chosen from the HLA-DR1, HLA-DR3, HLA-DR4, HLA-DR7, HLA-DR11, HLA-DR13, HLA-DR15, HLA-DRB3, HLA-DRB4, HLA-DRB5, HLA-DP401 and HLA-DP402 molecules, selected from the group consisting of: a) the sequences of 11 to 30 consecutive amino acids of the human cyclin B1 sequence of SEQ ID No. 1 comprising at least residues 115-123, 146-154, 166-174, 172-180, 187-195, 191-199, 197-205, 239-247, 266-274, 271-279, 277-285, 293-301, 329-337, 361-369 or 387-395 of said sequence SEQ ID No. 1, and b) the sequences of 11 to 30 amino acids having at least 70% identity with a sequence in a). 
     
     
         8 : The mixture of peptides as claimed in  claim 7 , wherein said second peptide is selected from the group consisting of the sequences SEQ ID Nos. 25, 29, 32, 34, 36, 37, 38, 47, 53, 55, 56, 59, 66, 72 and 78. 
     
     
         9 : The mixture of peptides as claimed in  claim 4 , wherein said peptides are linked to one another in the form of a multi-epitope polypeptide. 
     
     
         10 : A modified peptide or polypeptide derived from the peptide as claimed in  claim 1  through the introduction of a chemical modification, said modified peptide comprising at least one human cyclin B1 CD4 +  T epitope which is immunodominant in vitro and being capable of stimulating a specific human CD4 +  T lymphocyte response. 
     
     
         11 : An isolated polynucleotide encoding a peptide as claimed in  claim 1 . 
     
     
         12 : A recombinant expression vector comprising a polynucleotide as claimed in  claim 11 . 
     
     
         13 : An immunogenic or vaccine composition comprising at least one peptide as claimed in  claim 1 . 
     
     
         14 : A method of treating a cancer associated with the overexpression of cyclin B1 with prophylactic or curative immunotheraphy in a subject, the method comprising administering to the subject a composition comprising the peptide of  claim 1 .