ArticleJournal of immunology (Baltimore, Md. : 1950)2015
Antibody stabilization of peptide-MHC multimers reveals functional T cells bearing extremely low-affinity TCRs.
Article in Journal of immunology (Baltimore, Md. : 1950), 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
43 citing papers in PubMed, 57 citations in OpenAlex.
- Autoreactive T effector memory differentiation mirrors β cell function in type 1 diabetes.The Journal of clinical investigation · 2018Trial
- Computational design and translational development of TCR-mimic scFv antibodies for peptide-HLA neoantigen targeting.npj drug discovery · 2026Review
- Optimized procurement of cancer-reactive T-cell receptors from clinical samples.Immunotherapy advances · 2026Article
- Regulatory T cells constrain T cells of shared specificity to enforce tolerance during infection.Science (New York, N.Y.) · 2025Article
- Immunogenicity of Non-Mutated Ovarian Cancer-Specific Antigens.Current oncology (Toronto, Ont.) · 2024Article
- What's the Catch? The Significance of Catch Bonds in T Cell Activation.Journal of immunology (Baltimore, Md. : 1950) · 2023Review
- The influence of three-dimensional structure on naïve T cell homeostasis and aging.Frontiers in aging · 2022Article
- Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition).European journal of immunology · 2021Review
- Altered selection on a single self-ligand promotes susceptibility to organ-specific T cell infiltration.The Journal of experimental medicine · 2021Article
- MHC multimer: A Molecular Toolbox for Immunologists.Molecules and cells · 2021Review
- Molecular characterization of HLA class II binding to the LAG-3 T cell co-inhibitory receptor.European journal of immunology · 2021Article
- Role of CD4+ T Cells in the Control of Viral Infections: Recent Advances and Open Questions.International journal of molecular sciences · 2021Review
- Relationship of 2D Affinity to T Cell Functional Outcomes.International journal of molecular sciences · 2020Review
- Tracking immunodynamics by identification of S-GJournal of autoimmunity · 2020Article
- CD4Cell reports · 2020Article
- Circulating β cell-specific CD8Clinical and experimental immunology · 2020Article
- Genome-wide CRISPR-Cas9 screening reveals ubiquitous T cell cancer targeting via the monomorphic MHC class I-related protein MR1.Nature immunology · 2020Article
- GPU-Accelerated Discovery of Pathogen-Derived Molecular Mimics of a T-Cell Insulin Epitope.Frontiers in immunology · 2020Article
- Human leukocyte antigen (HLA) class II peptide flanking residues tune the immunogenicity of a human tumor-derived epitope.The Journal of biological chemistry · 2019Article
- Article
Corrections and comments
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Authors and funding
16 authors at 4 institutions in 4 countries.
Funding
Abstract
Fluorochrome-conjugated peptide-MHC (pMHC) multimers are commonly used in combination with flow cytometry for direct ex vivo visualization and characterization of Ag-specific T cells, but these reagents can fail to stain cells when TCR affinity and/or TCR cell-surface density are low. pMHC multimer staining of tumor-specific, autoimmune, or MHC class II-restricted T cells can be particularly challenging, as these T cells tend to express relatively low-affinity TCRs. In this study, we attempted to improve staining using anti-fluorochrome unconjugated primary Abs followed by secondary staining with anti-Ab fluorochrome-conjugated Abs to amplify fluorescence intensity. Unexpectedly, we found that the simple addition of an anti-fluorochrome unconjugated Ab during staining resulted in considerably improved fluorescence intensity with both pMHC tetramers and dextramers and with PE-, allophycocyanin-, or FITC-based reagents. Importantly, when combined with protein kinase inhibitor treatment, Ab stabilization allowed pMHC tetramer staining of T cells even when the cognate TCR-pMHC affinity was extremely low (KD >1 mM) and produced the best results that we have observed to date. We find that this inexpensive addition to pMHC multimer staining protocols also allows improved recovery of cells that have recently been exposed to Ag, improvements in the recovery of self-specific T cells from PBMCs or whole-blood samples, and the use of less reagent during staining. In summary, Ab stabilization of pMHC multimers during T cell staining extends the range of TCR affinities that can be detected, yields considerably enhanced staining intensities, and is compatible with using reduced amounts of these expensive reagents.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.