Engineered T-cell receptor tetramers bind MHC-peptide complexes with high affinity.
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.
Subbramanian
Ramu A
RA
Division of Viral Pathogenesis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, Massachusetts 02215, USA.
Moriya
Chikaya
C
Martin
Kristi L
KL
Peyerl
Fred W
FW
Hasegawa
Atsuhiko
A
Naoi
Akira
A
Chhay
Heng
H
Autissier
Patrick
P
Gorgone
Darci A
DA
Lifton
Michelle A
MA
Kuus-Reichel
Kristine
K
Schmitz
Jörn E
JE
Letvin
Norman L
NL
Kuroda
Marcelo J
MJ
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*
D000936:Antigen-Antibody Complex; D046912:Multiprotein Complexes; D016693:Receptors, Antigen, T-Cell, alpha-beta