Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers
Abstract
The use of fluorescently labeled major histocompatibility complex multimers has become an essential technique for analyzing disease- and therapy-induced T-cell immunity. Whereas classical major histocompatibility complex multimer analyses are well-suited for the detection of immune responses to a few epitopes, limitations on human-subject sample size preclude a comprehensive analysis of T-cell immunity. To address this issue, we developed a combinatorial encoding strategy that allows the parallel detection of a multitude of different T-cell populations in a single sample. Detection of T cells from peripheral blood by combinatorial encoding is as efficient as detection with conventionally labeled multimers but results in a substantially increased sensitivity and, most notably, allows comprehensive screens to be performed. We obtained proof of principle for the feasibility of large-scale screening of human material by analysis of human leukocyte antigen A3-restricted T-cell responses to known and potential melanoma-associated antigens in peripheral blood from individuals with melanoma.
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Formation of pMHC complexes was validated in a bead-based flow cytometry assay.
“pMHC complexes were refolded with the easYmer technology (easYmer kit, ImmuneAware Aps), and complex formation was validated in a bead-based flow cytometry assay according to the manufacturer's instructions 49,50 .”
From BNT162b2 vaccine induces neutralizing antibodies and poly-specific T cells in humans (2021) · cites this paper as [49]
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- Authors
- Hadrup, S. R., Bakker, A. H., Shu, C. J., Andersen, R. S., van Veluw, J., Hombrink, P., Castermans, E., thor Straten, P., Blank, C., Haanen, J. B., Heemskerk, M. H., & Schumacher, T. N.
- Affiliations
- The Netherlands Cancer Institute (NL)
- Herlev Hospital (DK)
- Leiden University Medical Center (NL)
- Corresponding author
- Schumacher, Ton N
- ORCID
- Venue
- Nature Methods
- Published
- 2009-06-21
- Volume
- 6
- Issue
- 7
- Pages
- 520-526
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- OpenAlex
- W2139517175
- Cited by
- 306
- References
- 20
- Citations by year
- 2026: 7 · 2025: 12 · 2024: 23 · 2023: 15 · 2022: 14 · 2021: 25 · 2020: 16 · 2019: 16 · 2018: 16 · 2017: 17 · 2016: 28 · 2015: 14 · 2014: 16 · 2013: 23 · 2012: 23
- FWCI
- 9.06
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- ORBi (University of Liège) (submittedVersion)
- Research at the University of Copenhagen (University of Copenhagen) (submittedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › vaccines and immunoinformatics approaches
- Keywords
- peptide-MHC tetramers, antigen-specific T cell responses, parallel detection, HLA-A3, melanoma-associated antigens, melanoma, peripheral blood, detection sensitivity
- MeSH terms
- Epitopes, Antigens, Antigens, Neoplasm, Cell Separation, Fluorescent Dyes, Histocompatibility Antigens, Humans, Immunologic Techniques, Neoplasm Proteins, Peptides, Sensitivity and Specificity, T-Lymphocyte Subsets, Protein Structure, Quaternary, Nanotechnology, Quantum Dots, Melanoma-Specific Antigens
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