NY-ESO-1 peptide derivatives, and uses thereof

Abstract

The invention relates to variant peptides which bind to HLA molecules, leading to lysis of cells via cytolytic T cell lines. The variants are based upon NY-ESO-1 peptides. The peptides can be incorporated into immune tetramers, which are useful as T cell sorters.

We claim:  
     
         1 . An isolated nonapeptide of formula  
       SLLMWITQX  
       (SEQ ID NO: 10) wherein X is any amino acid but cysteine, wherein said nonapeptide binds to an HLA molecule and provokes lysis by cytolytic T cells.  
     
     
         2 . The isolated nonapeptide of  claim 1 , wherein X is an amino acid having a non-polar side chain.  
     
     
         3 . The isolated nonapeptide of  claim 2 , wherein said amino acid is Ala, Val, Leu, Ile, Pro, Phe, Met, Trp or Gly.  
     
     
         4 . The isolated nonapeptide of  claim 1 , wherein said amino acid is Ala, Val, Ile or Leu.  
     
     
         5 . The isolated complex of  claim 18 , wherein said complex is a tetramer.  
     
     
         6 . The isolated tetramer of  claim 3 , wherein X in SEQ ID NO: 10 is A, V, I or L.  
     
     
         7 . Composition useful in provoking a cytolytic T cell response comprising the isolated nonapeptide of  claim 1 , and an adjuvant.  
     
     
         8 . An isolated nucleic acid molecule consisting of a nucleotide sequence which encodes the nonapeptide of  claim 1 .  
     
     
         9 . The isolated nucleic acid molecule of  claim 1 , wherein said nonapeptide consists of the amino acid sequence set forth at SEQ ID NO. 6, 7, 8, or 12.  
     
     
         10 . Expression vector comprising a plurality of nucleotide sequences which encode peptides which bind to MHC molecules, wherein at least one of said peptides is the peptide of  claim 1 .  
     
     
         11 . The expression vector of  claim 10 , wherein said peptide consists of the amino acid sequence of SEQ ID NO: 6, 7, 8 or 12.  
     
     
         12 . Recombinant cell transformed or transfected with the isolated nucleic acid molecule of  claim 8 .  
     
     
         13 . Recombinant cell transformed or transfected with the expression vector of  claim 10 .  
     
     
         14 . Method for determining if a cell presents an HLA-A2 molecule on its surface comprising contacting a sample containing said cell with the peptide of  claim 1 , and determining binding therebetween, said binding being indicative of HLA-A2 on the surface of said cell.  
     
     
         15 . The composition of  claim 7 , further comprising at least one additional peptide.  
     
     
         16 . Isolated polytope molecule, at least a portion of which comprises the amino acid sequence of  claim 1 .  
     
     
         17 . A method for determining if a cytolytic T cell specific to complexes of an HLA-A2 molecule and a peptide is present in a sample, comprising admixing said sample with an HLA-A2 molecule and the nonapeptide of  claim 1 , and determining interaction between said cytolytic T cell, and complexes of HLA-A2 molecule and said peptide to determine specificity of said cytolytic T cell.  
     
     
         18 . An isolated complex useful in isolating a cytolytic T cell, comprising a first and second binding partner which are specific to each other, wherein said second binding partner is bound to a plurality of complexes of an HLA-A2 molecule, a β2 microglobulin molecule, and the nonapeptide of  claim 1 .  
     
     
         19 . The isolated complex of  claim 18 , wherein said peptide is the peptide of SEQ ID NO: 6, 7, 8 or 12.  
     
     
         20 . The isolated complex of  claim 18 , further comprising a label.  
     
     
         21 . The isolated complex of  claim 18 , wherein said first binding partner is avidin and said second binding partner is, biotin.  
     
     
         22 . The isolated complex of  claim 21 , comprising complexes of MHC molecule, β2 microglobulin and peptide.  
     
     
         23 . A method for identifying or isolating cytolytic T cells in a sample, comprising admixing said sample with the complex of  claim 18 , and identifying or isolating cytolytic T cells which bind thereto.  
     
     
         24 . A method for monitoring status of a tumor, comprising contacting a sample taken from a patient with a tumor with the isolated complex of  claim 18  to determine cytolytic T cells in said sample, and comparing a value obtained to a previously determined value to determine status of said tumor.  
     
     
         25 . An isolated decapeptide of formula:  
       SLLMWITQXX  
       (SEQ ID NO: 18), wherein, the first X is cysteine or alanine, and the second X is any amino acid.  
     
     
         26 . The isolated decapeptide of  claim 25 , wherein the second X is Phe, Ile, Val or Leu.  
     
     
         27 . The isolated decapeptide of  claim 25 , wherein the first X is cysteine and the second X is Phe, Ile, Val or Leu.  
     
     
         28 . The isolated decapeptide of  claim 25 , wherein the first X is alanine and the second X is Phe, Ile, Val or Leu.  
     
     
         29 . The isolated decapeptide of  claim 27 , wherein the second X is Phe (SEQ. ID NO: 11), or Ile (SEQ ID NO: 14).  
     
     
         30 . Composition useful in provoking a cytolytic T cell response comprising the isolated decapeptide of  claim 25 , and an adjuvant.  
     
     
         31 . An isolated nucleic acid molecule consisting of a nucleotide sequence which encodes the isolated decapeptide  claim 25 .  
     
     
         32 . The isolated nucleic acid molecule of  claim 31 , wherein said decapeptide consists of the amino acid sequence set forth at SEQ. ID NO: 11, 13, 14, 15, 16 or 17.  
     
     
         33 . Expression vector comprising a plurality of nucleotide sequences which encode peptides which bind to MHC molecules, wherein at least one of said peptides is the peptide of  claim 25 .  
     
     
         34 . The expression vector of  claim 33 , wherein said peptide is the peptide of SEQ. ID NO: 11, 13, 14, 15, 16 or 17.  
     
     
         35 . Recombinant cell transformed or transfected with the isolated nucleic acid molecule of  claim 31 .  
     
     
         36 . Recombinant cell transformed or transfected with the expression vector of  claim 33 .  
     
     
         37 . A method for determining if a cell presents an HLA-A2 molecule on its surface comprising contacting a sample containing said cell with the decapeptide of  claim 25 , and determining binding therebetween, said binding being indicative of HLA-A2 on the surface of said cell.  
     
     
         38 . The composition of  claim 30 , further comprising at least one additional peptide.  
     
     
         39 . Isolated polytope molecule, at least a portion of which comprises the amino acid sequence of  claim 25 .  
     
     
         40 . A method for determining if a cytolytic T cell specific to complexes of an HLA-A2 molecule and a peptide is present in a sample, comprising admixing said sample with an HLA-A2 molecule and the decapeptide of  claim 25  and determining interaction between said cytolytic T cell, and complexes of HLA-A2 molecule and said peptide to determine specificity of said cytolytic T cell.  
     
     
         41 . An isolated complex useful in isolating a cytolytic T cell, comprising a first and second binding partner which are specific to each other, wherein said second binding partner is bound to complexes if an HLA-A2 molecule, a β2 microglobulin molecule, and the decapeptide of  claim 25 .  
     
     
         42 . The isolated complex of  claim 41 , wherein said complex is a tetramer.  
     
     
         43 . The isolated complex of  claim 41 , wherein said decapeptide has the amino acid sequence of SEQ. ID NO: 11, 13, 14, 15, 16 or 17.  
     
     
         44 . The isolated complex of  claim 41 , further comprising a label.  
     
     
         45 . The isolated complex of  claim 41 , wherein said first binding partner is avidin and said second binding partner is biotin.  
     
     
         46 . The isolated complex of  claim 45 , comprising complexes of MHC molecule, β2 microglobulin and decapeptide.  
     
     
         47 . A method for identifying or isolating cytolytic T cells in a sample, comprising admixing said sample with the complex of  claim 41 , and identifying or isolating cytolytic T cells which bind thereto.  
     
     
         48 . A method for monitoring status of a tumor, comprising contacting a sample taken from a patient with a tumor with the isolated complex of  claim 41  to determine cytolytic T cells in said sample, and comparing a value obtained to a previously determined value to determine status of said tumor.