title: Engineered T-cell receptor tetramers bind MHC-peptide complexes with high affinity.
abstract: In this study we extend tetramerization technology to T-cell receptors (TCRs). We identified TCR alpha beta pairs in the absence of accessory molecules, ensuring isolation of high-affinity TCRs that maintain stable binding characteristics after tetramerization. Subtle changes in cognate peptide levels bound to the class I molecule were accurately reflected by parallel changes in the mean fluorescence intensity of cells that bound TCR tetramers, allowing us to accurately assess the binding affinity of a panel of peptides to major histocompatibility complex (MHC) class I. Using a TCR tetramer specific for the Mamu-A(*)01 allele, we identified animals expressing this restricting class I allele from a large cohort of outbred rhesus macaques. TCR tetramers should facilitate analysis of the MHC-peptide interface and, more generally, the design of immunotherapeutics and vaccines.
authors.affiliation: Division of Viral Pathogenesis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, Massachusetts 02215, USA.
authors.forename: Ramu A
authors.initials: RA
authors.lastname: Subbramanian
authors.forename: Chikaya
authors.initials: C
authors.lastname: Moriya
authors.forename: Kristi L
authors.initials: KL
authors.lastname: Martin
authors.forename: Fred W
authors.initials: FW
authors.lastname: Peyerl
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Hasegawa
authors.forename: Akira
authors.initials: A
authors.lastname: Naoi
authors.forename: Heng
authors.initials: H
authors.lastname: Chhay
authors.forename: Patrick
authors.initials: P
authors.lastname: Autissier
authors.forename: Darci A
authors.initials: DA
authors.lastname: Gorgone
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Lifton
authors.forename: Kristine
authors.initials: K
authors.lastname: Kuus-Reichel
authors.forename: Jörn E
authors.initials: JE
authors.lastname: Schmitz
authors.forename: Norman L
authors.initials: NL
authors.lastname: Letvin
authors.forename: Marcelo J
authors.initials: MJ
authors.lastname: Kuroda
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis; D000937:Antigen-Antibody Reactions / Q000276:immunology; D002478:Cells, Cultured; D019281:Dimerization; D016231:Fluorescence Polarization Immunoassay / Q000379:methods*; D005805:Genes, MHC Class I / Q000235:genetics / Q000276:immunology*; D008253:Macaca mulatta; D046912:Multiprotein Complexes / Q000235:genetics / Q000276:immunology; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000936:Antigen-Antibody Complex; D046912:Multiprotein Complexes; D016693:Receptors, Antigen, T-Cell, alpha-beta