Cd4+ Survivin Epitopes and Uses Thereof

Abstract

The invention concerns CD4+ T survivin epitopes and their vaccine and diagnostic uses.

1 . A peptide derived from the alpha-isoform of survivin, for use as an antigen in the prophylactic or therapeutic immunization against cancer or the diagnosis, prognosis or therapeutic monitoring of cancer, said peptide being selected from the group consisting of:
 a) the peptides of 13 to 18 consecutive amino acids located between positions 17 and 34 of the alpha-isoform of survivin,   b) the peptides of 13 to 30 consecutive amino acids located between positions 84 and 113 of the alpha-isoform of survivin,   c) the peptides of 13 to 21 consecutive amino acids located between positions 122 and 142 of the alpha-isoform of survivin, and   d) the variants of the peptides defined in a), b) or c),   
       said peptides in a), b) or c), or variants in d), having a binding activity with respect to at least one HLA II molecule predominant in the Caucasian population, of less than 1000 nM, and being capable of inducing survivin-specific CD4+ T lymphocytes. 
     
     
         2 . The peptide as claimed in  claim 1 , characterized in that said HLA II molecule predominant in the Caucasian population is selected from the group consisting of: HLA-DR 1, HLA-DR3, HLA-DR4, HLA-DR7, HLA-DR11, HLA-DR13, HLA-DR15, HLA-DRB3, HLA-DRB4, HLA-DRB5 and HLA-DP4. 
     
     
         3 . The peptide as claimed in  claim 2 , characterized in that said HLA II molecule is encoded, respectively, by the HLA alleles DRB1*0101, DRB1*0301, DRB 1*0401, DRB 1*0701, DRB1*1101, DRB1*1301, DRB1*1501, DRB3*0101, DRB4*0101, DRB5*0101, DP*0401 and DP*0402. 
     
     
         4 . The peptide as claimed in  claim 1 , characterized in that it is capable of being presented by at least three HLA II molecules predominant in the Caucasian population. 
     
     
         5 . The peptide as claimed in  claim 3 , characterized in that it is capable of being presented by at least one HLA-DP403 or HLA-DP402 molecule. 
     
     
         6 . The peptide as claimed in  claim 4 , characterized in that it is selected from the group consisting of the peptides of 15 amino acids located between the following positions of the alpha-isoform of survivin: 17 to 31, 19 to 33, 20 to 34, 84 to 98, 90 to 104, 91 to 105,93 to 107, 96 to 110, 99 to 113, 122 to 136 and 128 to 142. 
     
     
         7 . The peptide as claimed in  claim 6 , characterized in that it is capable of being presented by at least four HLA II molecules predominant in the Caucasian population, and in that it is selected from the group consisting of the peptides: 17-31, 19-33, 20-34, 84-98, 90-104, 91-105, 93-107, 96-110 and 128-142. 
     
     
         8 . The peptide as claimed in  claim 7 , characterized in that it is capable of being presented by an HLA-DP401 or HLA-DP402 molecule, and in that it is selected from the group consisting of the peptides: 17-31, 19-33, 20-34, 84-98, 90-104, 91-105, 93-107 and 96-110. 
     
     
         9 . The peptide as claimed in  claim 6 , characterized in that it is selected from the group consisting of the sequences SEQ ID Nos: 5, 6, 7, 17, 19, 20, 21, 23, 24, 27 and 28. 
     
     
         10 . The peptide as claimed in  claim 1 , characterized in that it is a variant of a peptide as defined in  claim 1 , obtained by substitution of at least one of the residues P1, P6 and/or P9 for anchoring to the HLA II molecules, with an aromatic or hydrophobic amino acid, of the residue P6 with a cysteine (C), of the residue P9 with C, D, Q, S, T or E, and/or of the residue P4 with another natural or synthetic amino acid. 
     
     
         11 . The peptide as claimed in  claim 1  characterized in that it has a sequence of 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 amino acids. 
     
     
         12 . The peptide as claimed in  claim 1 , characterized in that it is labeled or complexed, 
     
     
         13 . The peptide as claimed in  claim 12 , characterized in that it is complexed with a labeled HLA II molecule. 
     
     
         14 . A polyepitopic fragment comprising the concatenation of at least two identical or different epitopes, at least one of which is a peptide as defined in  claim 1  for use as an antigen in the prophylactic or therapeutic vaccination against cancer or the diagnosis, prognosis or therapeutic monitoring of cancer. 
     
     
         15 . The polyepitopic fragment as claimed in  claim 14 , characterized in that it comprises the concatenation of at least one peptide as defined in  claim 1  and of at least one survivin CD8+ T epitope and/or a universal CD4+ T epitope, 
     
     
         16 . The polyepitopic fragment as claimed in  claim 15 , characterized in that the CD8+ T epitope is chosen from the sequences SEQ ID Nos: 37 to 39. 
     
     
         17 . A fusion protein consisting of a protein or a protein fragment, fused to a peptide as defined in  claim 1 . 
     
     
         18 . The fusion protein as claimed in  claim 17 , characterized in that said protein or said fragment is selected from the group consisting of an alpha- or beta-chain of an HLA II molecule, a fragment of said chain corresponding to a soluble HLA II molecule and a sequence for targeting to the endosome. 
     
     
         19 . The fusion protein as claimed in  claim 18 , characterized in that said targeting sequence is derived from the invariable chain Ii or from the LAMP-1 protein. 
     
     
         20 . A lipopeptide obtained by addition of a lipid to an α-amino function or a reactive function of the side chain of a peptide as defined in  claim 1 . 
     
     
         21 . An expression vector comprising a polynucleotide encoding a peptide as defined in  claim 1 , under the control of appropriate regulatory sequences, for use as an antigen in the prophylactic or therapeutic immunization against cancer or the diagnosis, prognosis or therapeutic monitoring of cancer. 
     
     
         22 . An immunogenic or vaccinal composition, characterized in that it comprises at least one peptide as claimed in  claim 1 , and a pharmaceutically acceptable carrier, a carrier substance or an adjuvant. 
     
     
         23 . The immunogenic or vaccinal composition as claimed in  claim 22 , characterized in that it comprises the at least one peptide and one peptide including a survivin CD8+ T epitope, in the form of a mixture of peptides, of a polyepitopic fragment and/or of an expression vector encoding said peptides or said fragment. 
     
     
         24 . The immunogenic or vaccinal composition as claimed in  claim 23 , characterized in that said CD8+ T epitope is chosen from the sequences SEQ ID Nos: 37 to 39. 
     
     
         25 . The immunogenic or vaccinal composition according to  claim 22 , characterized in that it comprises at least two different peptides, selected from the group consisting of one of the following combinations:
 peptide 17-31 and at least one of peptides 19-33, 90-104 or 128-142,   peptide 19-33 and peptide 96-110,   peptide 90-104 and peptide 17-31,   peptide 96-110 and peptide 90-104, and   peptides 93-107 and 128-142, and at least one of peptides 17-31, 19-33, 96-110 or 90-104.   
     
     
         26 . The immunogenic composition as claimed in  claim 22 , characterized in that it comprises a universal CD4+ T epitope. 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . An in vitro method for diagnosing, evaluating the prognosis of or monitoring the treatment of a cancer in an individual, characterized in that it comprises:
 bringing a biological sample from said individual into contact, in vitro, with a peptide as claimed in  claim 1 , and   detecting survivin-specific CD4+ T lymphocytes by any appropriate means,   
     
     
         30 . A method for sorting survivin-specific CD4+ T lymphocytes, characterized in that it comprises at least the following steps:
 bringing a cell sample into contact, in vitro, with labeled HLA II/peptide multimeric complexes formed by the binding of soluble HLA II molecules with at least one peptide as claimed in  claim 1 , and   sorting the cells bound to said HLA II/peptide complexes,   
     
     
         31 . The peptide as defined in  claim 1 , with the exclusion of the peptide located between positions 89 and 101 of the alpha-isoform of survivin. 
     
     
         32 . A polyepitopic fragment comprising the concatenation of at least two identical or different epitopes, at least one of which is a peptide as defined in  claim 1 . 
     
     
         33 . A fusion protein consisting of a protein or a protein fragment, fused to a peptide as defined in  claim 1  or a polyepitopic fragment as claimed in  claim 32 . 
     
     
         34 . A lipopeptide obtained by the addition of a lipid to an α-amino function or a reactive function of the side chain of a peptide as defined in  claim 1  or of a polyepitopic fragment as claimed in  claim 32 . 
     
     
         35 . A polynucleotide encoding a peptide as claimed in  claim 31 , a polyepitopic fragment as claimed in  claim 32  or a fusion protein as claimed in  claim 33 . 
     
     
         36 . An expression vector comprising a polynucleotide as claimed in  claim 35  under the control of appropriate regulatory sequences for transcription and, optionally, for translation. 
     
     
         37 . A host cell modified with a polynucleotide as claimed in  claim 35  or an expression vector as claimed in  claim 36 .