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Engineered T-cell receptor tetramers bind MHC-peptide complexes with high affinity

Nature Biotechnology volume 22, pages 1429–1434 (2004)Cite this article

Abstract

In this study we extend tetramerization technology to T-cell receptors (TCRs). We identified TCR αβ 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.

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Figure 1: A schematic representation of the screening of TCR αβ chain pairs using a peptide-specific TL8-Mamu-A*01 tetramer complex.
Figure 2: Tat-TL8 TCR tetramer binds specifically to TL8 peptide–pulsed Mamu-A*01–expressing cells.
Figure 3: Changes in the mean fluorescence intensity of TCR-tetramer staining reflect changes in bound levels of the cognate Tat-TL8 peptide.
Figure 4: TCR tetramer identifies animals that express the Mamu-A*01 allele in a cohort of outbred rhesus macaques.

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Acknowledgements

We thank Rudolf P. Bohm, Jr. and Andrew A. Lackner from the Tulane National Primate Research Center (TNPRC) for providing monkey blood samples; and David I. Watkins from the Wisconsin Regional Primate Research Center for the Mamu-A*01 expressing 721.221 cell line used in this study. We thank Carol Lord, Karen Hershberger and William Charini for helpful discussions. This work was supported by National Institutes of Health grants RO1 AI48400 (M.J.K.), AI 48394 (J.E.S.), AI20279 (N.L.L.), AI85343 (N.L.L.), the Dana-Farber Cancer Institute/Beth Israel Deaconess Medical Center/Children's Hospital Center for AIDS Research Grant P30 AI28691 (M.J.K.) and the base grant to the TNPRC RR00168.

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Author notes

  1. Ramu A Subbramanian, Chikaya Moriya and Kristi L Martin: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Viral Pathogenesis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, 02215, Massachusetts, USA

    Ramu A Subbramanian, Chikaya Moriya, Kristi L Martin, Fred W Peyerl, Atsuhiko Hasegawa, Akira Naoi, Heng Chhay, Patrick Autissier, Darci A Gorgone, Michelle A Lifton, Jörn E Schmitz, Norman L Letvin & Marcelo J Kuroda

  2. Immunomics Operations, Beckman Coulter, San Diego, 92121, California, USA

    Kristine Kuus-Reichel

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Correspondence to Marcelo J Kuroda.

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Supplementary information

Supplementary Fig. 1

Binding of soluble peptide/MCH complexes to the TCR on S2 cells. (PDF 37 kb)

Supplementary Table 1

Peptide binding to Mamu-A*01 (PDF 17 kb)

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Subbramanian, R., Moriya, C., Martin, K. et al. Engineered T-cell receptor tetramers bind MHC-peptide complexes with high affinity. Nat Biotechnol 22, 1429–1434 (2004). https://doi.org/10.1038/nbt1024

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