title: High-affinity TCRs generated by phage display provide CD4+ T cells with the ability to recognize and kill tumor cell lines.
abstract: We examined the activity of human T cells engineered to express variants of a single TCR (1G4) specific for the cancer/testis Ag NY-ESO-1, generated by bacteriophage display with a wide range of affinities (from 4 microM to 26 pM). CD8(+) T cells expressing intermediate- and high-affinity 1G4 TCR variants bound NY-ESO-1/HLA-A2 tetramers with high avidity and Ag specificity, but increased affinity was associated with a loss of target cell specificity of the TCR gene-modified cells. T cells expressing the highest affinity TCR (K(D) value of 26 pM) completely lost Ag specificity. The TCRs with affinities in the midrange, K(D) 5 and 85 nM, showed specificity only when CD8 was absent or blocked, while the variant TCRs with affinities in the intermediate range-with K(D) values of 450 nM and 4 microM-demonstrated Ag-specific recognition. Although the biological activity of these two relatively low-affinity TCRs was comparable to wild-type reactivity in CD8(+) T cells, introduction of these TCR dramatically increased the reactivity of CD4(+) T cells to tumor cell lines.
authors.affiliation: Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Yangbing
authors.initials: Y
authors.lastname: Zhao
authors.forename: Alan D
authors.initials: AD
authors.lastname: Bennett
authors.forename: Zhili
authors.initials: Z
authors.lastname: Zheng
authors.forename: Qiong J
authors.initials: QJ
authors.lastname: Wang
authors.forename: Paul F
authors.initials: PF
authors.lastname: Robbins
authors.forename: Lawrence Y L
authors.initials: LY
authors.lastname: Yu
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.forename: Peter E
authors.initials: PE
authors.lastname: Molloy
authors.forename: Steven M
authors.initials: SM
authors.lastname: Dunn
authors.forename: Bent K
authors.initials: BK
authors.lastname: Jakobsen
authors.forename: Steven A
authors.initials: SA
authors.lastname: Rosenberg
authors.forename: Richard A
authors.initials: RA
authors.lastname: Morgan
mesh_terms: D000595:Amino Acid Sequence; D000938:Antigen-Presenting Cells / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002470:Cell Survival; D002478:Cells, Cultured; D003429:Cross Reactions / Q000276:immunology; D006649:Histocompatibility Antigens / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000276:immunology* / Q000378:metabolism / Q000473:pathology*; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: D006649:Histocompatibility Antigens; D019151:Peptide Library; D011948:Receptors, Antigen, T-Cell