Changing the peptide specificity of a human T-cell receptor by directed evolution.
Binding of a T-cell receptor (TCR) to a peptide/major histocompatibility complex is the key interaction involved in antigen specificity of T cells. The recognition involves up to six complementarity determining regions (CDR) of the TCR. Efforts to examine the structural basis of these interactions and to exploit them in adoptive T-cell therapies has required the isolation of specific T-cell clones and their clonotypic TCRs. Here we describe a strategy using in vitro-directed evolution of a single TCR to change its peptide specificity, thereby avoiding the need to isolate T-cell clones. The human TCR A6, which recognizes the viral peptide Tax/HLA-A2, was converted to TCR variants that recognized the cancer peptide MART1/HLA-A2. Mutational studies and molecular dynamics simulations identified CDR residues that were predicted to be important in the specificity switch. Thus, in vitro engineering strategies alone can be used to discover TCRs with desired specificities.
Department of Biochemistry, University of Illinois, 600 South Matthews Avenue, Urbana, Illinois 61801, USA.
Sheena N
SN
Smith
Center for Biophysics and Computational Biology, University of Illinois, Urbana, Illinois 61802, USA.
Yuhang
Y
Wang
Center for Biophysics and Computational Biology, University of Illinois, Urbana, Illinois 61802, USA.
Javier L
JL
Baylon
Department of Chemistry and Biochemistry and the Harper Cancer Research Institute, University of Notre Dame, 1234 Notre Dame Avenue, South Bend, Indiana 46557, USA.
Nishant K
NK
Singh
Department of Chemistry and Biochemistry and the Harper Cancer Research Institute, University of Notre Dame, 1234 Notre Dame Avenue, South Bend, Indiana 46557, USA.
Brian M
BM
Baker
1] Department of Biochemistry, University of Illinois, 600 South Matthews Avenue, Urbana, Illinois 61801, USA [2] Center for Biophysics and Computational Biology, University of Illinois, Urbana, Illinois 61802, USA.
Emad
E
Tajkhorshid
Department of Biochemistry, University of Illinois, 600 South Matthews Avenue, Urbana, Illinois 61801, USA.
David M
DM
Kranz
D000595:Amino Acid Sequence; D002999:Clone Cells; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000235:genetics*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics*; D019143:Evolution, Molecular; D015789:HLA-A2 Antigen / Q000737:chemistry / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D058965:MART-1 Antigen / Q000737:chemistry / Q000235:genetics; D008285:Major Histocompatibility Complex; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000235:genetics*
D022801:Complementarity Determining Regions; D000939:Epitopes; D015789:HLA-A2 Antigen; D058965:MART-1 Antigen; D009842:Oligopeptides; D011948:Receptors, Antigen, T-Cell; C518303:leucyl-leucyl-phenylalanyl-glycyl-tyrosyl-prolyl-valyl-tyrosyl-valine