Peptide-MHC Class I Tetramers Can Fail To Detect Relevant Functional T Cell Clonotypes and Underestimate Antigen-Reactive T Cell Populations.
Details
Serval ID
serval:BIB_5D3F0C23532F
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Peptide-MHC Class I Tetramers Can Fail To Detect Relevant Functional T Cell Clonotypes and Underestimate Antigen-Reactive T Cell Populations.
Journal
Journal of immunology
ISSN
1550-6606 (Electronic)
ISSN-L
0022-1767
Publication state
Published
Issued date
01/04/2018
Peer-reviewed
Oui
Volume
200
Number
7
Pages
2263-2279
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Publication Status: ppublish
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.
Keywords
CD8-Positive T-Lymphocytes/immunology, Cytomegalovirus/immunology, HLA-A2 Antigen/immunology, Herpesvirus 4, Human/immunology, Humans, Lymphocyte Activation/immunology, Lymphocytes, Tumor-Infiltrating/immunology, Melanoma/immunology, Orthomyxoviridae/immunology, Protein Binding/immunology, Protein Kinase Inhibitors/metabolism, RNA-Binding Proteins/immunology, Receptors, Antigen, T-Cell/immunology, Staining and Labeling/methods, Tumor Cells, Cultured
Pubmed
Web of science
Open Access
Yes
Create date
03/03/2018 15:08
Last modification date
20/08/2019 15:15