Affinity maturated t cell receptors and use thereof

Abstract

The present invention relates to methods and systems for increasing the affinity of a T cell receptor (TCR) to its ligand by subjecting the TCR gene to somatic hypermutation. The present invention further relates to use of affinity maturated TCRs to create T cells reactive against a selected antigen.

What is claimed is: 
     
         1 . A nucleic acid construct comprising:
 (i) a polycistronic nucleic acid construct comprising a nucleic acid sequence encoding a TCR comprising a TCR alpha chain and a TCR beta chain; and   (ii) a nucleic acid construct comprising a nucleic acid sequence encoding Activation Induced cytidine Deaminase (AID).   
     
     
         2 . The nucleic acid construct of  claim 1 , wherein said TCR is human TCR. 
     
     
         3 . The nucleic acid construct of  claim 1 , wherein said TCR comprises an N-terminal immunoglobulin variable domain, an immunoglobulin constant domain, a transmembrane-spanning region and a cytoplasmic region. 
     
     
         4 . The nucleic acid construct of  claim 1 , wherein said AID is human AID. 
     
     
         5 . The nucleic acid construct of  claim 1 , wherein the amino acid sequence of said AID is as set forth in SEQ ID NO: 4. 
     
     
         6 . The nucleic acid construct of  claim 1 , wherein said nucleic acid sequence encoding AID is as set forth in SEQ ID NO: 3. 
     
     
         7 . The nucleic acid construct of  claim 1 , wherein said construct comprises a regulatory element for directing expression of said TCR and said AID in a host cell. 
     
     
         8 . The nucleic acid construct of  claim 7 , wherein said nucleic acid construct encoding said TCR comprises a CMV promoter. 
     
     
         9 . The nucleic acid construct of  claim 7 , wherein said nucleic acid construct encoding AID comprises an inducible promoter. 
     
     
         10 . The s nucleic acid construct of  claim 9 , wherein said inducible promoter is a Tet-on promoter.