title: Single and dual amino acid substitutions in TCR CDRs can enhance antigen-specific T cell functions.
abstract: Single and dual amino acid substitution variants were generated in the TCR CDRs of three TCRs that recognize tumor-associated Ags. Substitutions that enhance the reactivity of TCR gene-modified T cells to the cognate Ag complex were identified using a rapid RNA-based transfection system. The screening of a panel of variants of the 1G4 TCR, that recognizes a peptide corresponding to amino acid residues 157-165 of the human cancer testis Ag NY-ESO-1 (SLLMWITQC) in the context of the HLA-A*02 class I allele, resulted in the identification of single and dual CDR3alpha and CDR2beta amino acid substitutions that dramatically enhanced the specific recognition of NY-ESO-1(+)/HLA-A*02(+) tumor cell lines by TCR gene-modified CD4(+) T cells. Within this group of improved TCRs, a dual substitution in the 1G4 TCR CDR3alpha chain was identified that enhanced Ag-specific reactivity in gene-modified CD4(+) and CD8(+) T cells. Separate experiments on two distinct TCRs that recognize the MART-1 27-35 (AAGIGILTV) peptide/HLA-A*02 Ag complex characterized single amino acid substitutions in both TCRs that enhanced CD4(+) T cell Ag-specific reactivity. These results indicate that simple TCR substitution variants that enhance T cell function can be identified by rapid transfection and assay techniques, providing the means for generating potent Ag complex-specific TCR genes for use in the study of T cell interactions and in T cell adoptive immunotherapy.
authors.affiliation: Surgery Branch, National Cancer Institute, National Institutes of Health, 10 Center Drive, Bethesda, MD 20892, USA. Paul_Robbins@nih.gov
authors.forename: Paul F
authors.initials: PF
authors.lastname: Robbins
authors.forename: Yong F
authors.initials: YF
authors.lastname: Li
authors.forename: Mona
authors.initials: M
authors.lastname: El-Gamil
authors.forename: Yangbing
authors.initials: Y
authors.lastname: Zhao
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Wargo
authors.forename: Zhili
authors.initials: Z
authors.lastname: Zheng
authors.forename: Hui
authors.initials: H
authors.lastname: Xu
authors.forename: Richard A
authors.initials: RA
authors.lastname: Morgan
authors.forename: Steven A
authors.initials: SA
authors.lastname: Feldman
authors.forename: Laura A
authors.initials: LA
authors.lastname: Johnson
authors.forename: Alan D
authors.initials: AD
authors.lastname: Bennett
authors.forename: Steven M
authors.initials: SM
authors.lastname: Dunn
authors.forename: Tara M
authors.initials: TM
authors.lastname: Mahon
authors.forename: Bent K
authors.initials: BK
authors.lastname: Jakobsen
authors.forename: Steven A
authors.initials: SA
authors.lastname: Rosenberg
mesh_terms: D019943:Amino Acid Substitution / Q000276:immunology; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D045744:Cell Line, Tumor; D022801:Complementarity Determining Regions / Q000235:genetics* / Q000276:immunology; D015234:HLA-A Antigens / Q000235:genetics / Q000276:immunology*; D015789:HLA-A2 Antigen; D006801:Humans; D058965:MART-1 Antigen; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; D022801:Complementarity Determining Regions; D015234:HLA-A Antigens; C524866:HLA-A*02 antigen; D015789:HLA-A2 Antigen; D058965:MART-1 Antigen; C548667:MLANA protein, human; D009363:Neoplasm Proteins; D011948:Receptors, Antigen, T-Cell