Peptide-MHC class 1 tetramers can fail to detect relevant functional T cell clonotypes and underestimate antigen-reactive T cell populations

Rius, Cristina, Attaf, Meriem, Tungatt, Katie, Bianchi, Valentina, Legut, Mateusz, Bovay, Amandine, Donia, Marco, Straten, Per thor, Peakman, Mark, Svane, Inge Marie, Ott, Sascha, Connor, Tom ORCID: https://orcid.org/0000-0003-2394-6504, Szomolay, Barbara ORCID: https://orcid.org/0000-0002-5375-5533, Dolton, Garry and Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135 2018. Peptide-MHC class 1 tetramers can fail to detect relevant functional T cell clonotypes and underestimate antigen-reactive T cell populations. Journal of Immunology 200 (7) , pp. 2263-2279. 10.4049/jimmunol.1700242

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Abstract

Peptide-MHC (pMHC) multimers, usually used as streptavidin-based tetramers, have transformed the study of Ag-specific T cells by allowing direct detection, phenotyping, and enumeration within polyclonal T cell populations. These reagents are now a standard part of the immunology toolkit and have been used in many thousands of published studies. Unfortunately, the TCR-affinity threshold required for staining with standard pMHC multimer protocols is higher than that required for efficient T cell activation. This discrepancy makes it possible for pMHC multimer staining to miss fully functional T cells, especially where low-affinity TCRs predominate, such as in MHC class II-restricted responses or those directed against self-antigens. Several recent, somewhat alarming, reports indicate that pMHC staining might fail to detect the majority of functional T cells and have prompted suggestions that T cell immunology has become biased toward the type of cells amenable to detection with multimeric pMHC. We use several viral- and tumor-specific pMHC reagents to compare populations of human T cells stained by standard pMHC protocols and optimized protocols that we have developed. Our results confirm that optimized protocols recover greater populations of T cells that include fully functional T cell clonotypes that cannot be stained by regular pMHC-staining protocols. These results highlight the importance of using optimized procedures that include the use of protein kinase inhibitor and Ab cross-linking during staining to maximize the recovery of Ag-specific T cells and serve to further highlight that many previous quantifications of T cell responses with pMHC reagents are likely to have considerably underestimated the size of the relevant populations.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Biosciences
Additional Information:	This article is distributed under the terms of the CC BY 4.0 Unported license.
Publisher:	American Association of Immunologists
ISSN:	0022-1767
Funders:	Wellcome Trust
Date of First Compliant Deposit:	21 February 2018
Date of Acceptance:	29 January 2018
Last Modified:	20 Sep 2024 16:43
URI:	https://orca.cardiff.ac.uk/id/eprint/109358

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