Detection of low avidity CD8+ T cell populations with coreceptor-enhanced peptide-major histocompatibility complex class I tetramers

Melenhorst, J. Joseph, Scheinberg, Phillip, Chattopadhyay, Pratip K., Lissina, Anna, Gostick, Emma, Cole, David ORCID: https://orcid.org/0000-0003-0028-9396, Wooldridge, Linda, van den Berg, Hugo A., Bornstein, Ethan, Hensel, Nancy F., Douek, Daniel C., Roederer, Mario, Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135, Barrett, A. John and Price, David ORCID: https://orcid.org/0000-0001-9416-2737 2008. Detection of low avidity CD8+ T cell populations with coreceptor-enhanced peptide-major histocompatibility complex class I tetramers. Journal of Immunological Methods 338 (1-2) , pp. 31-39. 10.1016/j.jim.2008.07.008

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Abstract

The development of soluble recombinant peptide-major histocompatibility complex class I (pMHCI) molecules conjugated in multimeric form to fluorescent labels has enabled the physical quantification and characterization of antigen-specific CD8+ T cell populations by flow cytometry. Several factors determine the binding threshold that enables visualization of cognate CD8+ T cells with these reagents; these include the affinity of the T cell receptor (TCR) for pMHCI antigen. Here, we show that multimers constructed from peptide-human leukocyte antigen (pHLA) A0201 monomers engineered in the heavy chain α2 domain to enhance CD8 binding (KD ≈ 85 μM) without impacting the TCR binding platform can detect cognate CD8+ T cells bearing low affinity TCRs that are not visible with the corresponding wildtype pHLA A0201 multimeric complexes. Mechanistically, this effect is mediated by a disproportionate enhancement of the TCR/pMHCI association rate. In direct ex vivo applications, these coreceptor-enhanced multimers exhibit faithful cognate binding properties; concomitant increases in background staining within the non-cognate CD8+ T cell population can be resolved phenotypically using polychromatic flow cytometry as a mixture of naïve and memory cells. These findings provide the first validation of a novel approach to the physical detection of low avidity antigen-specific CD8+ T cell populations; such coreceptor-enhanced multimeric reagents are likely to be useful in a multitude of settings for the detection of auto-immune, tumor-specific and cross-reactive CD8+ T cells.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Systems Immunity Research Institute (SIURI)
Subjects:	Q Science > QR Microbiology > QR180 Immunology
Uncontrolled Keywords:	peptide-major histocompatibility complex class I tetramer, polychromatic flow cytometry, T cell
Publisher:	Elsevier
ISSN:	0022-1759
Date of Acceptance:	1 July 2008
Last Modified:	15 May 2024 01:11
URI:	https://orca.cardiff.ac.uk/id/eprint/26007

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