title: CD8(-) T cell transfectants that express a high affinity T cell receptor exhibit enhanced peptide-dependent activation.
abstract: T cells are activated by binding of the T cell receptor (TCR) to a peptide-major histocompatibility complex (MHC) complex (pMHC) expressed on the surface of antigen presenting cells. Various models have predicted that activation is limited to a narrow window of affinities (or dissociation rates) for the TCR-pMHC interaction and that above or below this window, T cells will fail to undergo activation. However, to date there have not been TCRs with sufficiently high affinities in order to test this hypothesis. In this report we examined the activity of a CD8-negative T cell line transfected with a high affinity mutant TCR (K(D) = 10 nM) derived from cytotoxic T lymphocyte clone 2C by in vitro engineering. The results show that despite a 300-fold higher affinity and a 45-fold longer off-rate compared with the wild-type TCR, T cells that expressed the mutant TCRs were activated by peptide. In fact, activation could be detected at significantly lower peptide concentrations than with T cells that expressed the wild-type TCR. Furthermore, binding and functional analyses of a panel of peptide variants suggested that pMHC stability could account for apparent discrepancies between TCR affinity and T cell activity observed in several prior studies.
authors.affiliation: Department of Biochemistry, University of Illinois, Urbana, IL 61801, USA.
authors.forename: P D
authors.initials: PD
authors.lastname: Holler
authors.forename: A R
authors.initials: AR
authors.lastname: Lim
authors.forename: B K
authors.initials: BK
authors.lastname: Cho
authors.forename: L A
authors.initials: LA
authors.lastname: Rund
authors.forename: D M
authors.initials: DM
authors.lastname: Kranz
mesh_terms: D017951:Antigen Presentation / Q000276:immunology*; D016827:CD8 Antigens / Q000276:immunology*; D007376:Interleukin-2 / Q000096:biosynthesis; D007655:Ketoglutarate Dehydrogenase Complex / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology*; D008285:Major Histocompatibility Complex / Q000276:immunology; D016296:Mutagenesis; D009842:Oligopeptides / Q000276:immunology*; D010455:Peptides / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection
chemical_list: D016827:CD8 Antigens; D007376:Interleukin-2; D009842:Oligopeptides; D010455:Peptides; D016693:Receptors, Antigen, T-Cell, alpha-beta; C088354:p2Ca peptide; D007655:Ketoglutarate Dehydrogenase Complex