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Peptide-MHC Class I Tetramers Can Fail To Detect Relevant Functional T Cell
Clonotypes and Underestimate Antigen-Reactive T Cell Populations
#MMPMID29483360
Rius C
; Attaf M
; Tungatt K
; Bianchi V
; Legut M
; Bovay A
; Donia M
; Thor Straten P
; Peakman M
; Svane IM
; Ott S
; Connor T
; Szomolay B
; Dolton G
; Sewell AK
J Immunol
2018[Apr]; 200
(7
): 2263-2279
PMID29483360
show ga
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.
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