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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, Wooldridge, Linda, van den Berg, Hugo A., Bornstein, Ethan, Hensel, Nancy F., Douek, Daniel C., Roederer, Mario, Sewell, Andrew K., Barrett, A. John and Price, David 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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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: 07 Jul 2020 13:48

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