T cell receptor mutants

Abstract

A T cell receptor (TCR) having the property of binding to EVDPIGHLY HLA-A1 complex and comprising a specified wild type TCR which has specific mutations in the TCR alpha variable domain and/or the TCR beta variable domain to increase affinity. Such TCRs are useful for adoptive therapy.

1 . (canceled) 
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . A nucleic acid encoding a TCR having the property of binding to SEQ ID NO: 1-Human Leukocyte Antigen (HLA)-A1 complex and comprising a TCR α variable domain and a TCR β variable domain, wherein:
 the TCR α variable domain has the amino acid sequence from K1 to P114 of SEQ ID NO: 2 and the TCR β variable domain has the amino acid sequence from K1 to T112 of SEQ ID NO: 3 
 except that, in the TCR α variable domain, at least one of the following mutations is present, namely 
 50I is mutated to 50V; 
 51Q is mutated to 51R; 
 52S is mutated to 52P; 
 53S is mutated to 53Y; and/or 
 in the TCR β variable domain, at least one of the following mutations is present, namely 
 50F is mutated to 50T; 
 51S is mutated to 51D; 
 52E is mutated to 52M; 
 53T is mutated to 53L; 
 54Q is mutated to 54L. 
 
     
     
         7 . A cell comprising a TCR expression vector comprising the nucleic acid as claimed in  claim 6  in a single open reading frame or two distinct open reading frames. 
     
     
         8 . A cell comprising a first expression vector comprising a nucleic acid encoding the alpha chain of the TCR of  claim 6  and a second expression vector comprising a nucleic acid encoding the beta chain of the TCR of  claim 6 . 
     
     
         9 . A cell displaying on its surface a TCR encoded by the nucleic acid of  claim 6 . 
     
     
         10 . A cell as claimed in  claim 9  presenting a TCR having an α chain variable domain of SEQ ID NO: 8 and a β chain variable domain of SEQ ID NO: 3. 
     
     
         11 . A cell as claimed in  claim 9  presenting a TCR having an α chain variable domain of SEQ ID NO: 9 and a β chain variable domain of SEQ ID NO: 3. 
     
     
         12 . A cell as claimed in  claim 9  presenting a TCR having an α chain of SEQ ID NO: 2 and a β chain variable domain of SEQ ID No: 10. 
     
     
         13 . A cell as claimed in  claim 9  presenting a TCR having an α chain of SEQ ID NO: 2 and a β chain variable domain of SEQ ID NO: 11. 
     
     
         14 . A pharmaceutical composition comprising a plurality of cells as claimed in  claim 7  together with one or more pharmaceutically acceptable carriers or excipients. 
     
     
         15 . A nucleic acid as claimed in  claim 6  wherein the TCR comprises one of the α chain variable domain amino acid sequences SEQ ID NOs: 8 and 9. 
     
     
         16 . A nucleic acid as claimed in  claim 6  wherein the TCR comprises one of the β chain variable domain amino acid sequences SEQ ID NOs: 10 and 11. 
     
     
         17 . A nucleic acid as claimed in  claim 6  wherein the TCR also has an α chain TRAC constant domain and a β chain TRBC1 or TRBC2 constant domain, or has an α chain TRAC and β chain TRBC1 or TRBC2 constant domains modified by truncation or substitution to delete the native disulfide bond between Cys4 of exon 2 of TRAC and Cyst of exon 2 of TRBC1 or TRBC2. 
     
     
         18 . A nucleic acid as claimed in  claim 6  wherein the TCR is an αβ heterodimeric TCR, and has α and β chain constant domain sequences in which cysteine residues are substituted for Thr 48 of TRAC and Ser 57 of TRBC1 or TRBC2, the said cysteines forming a disulfide bond between the alpha and beta constant domains of the TCR.