Cited by 307T-cell and B-cell ImmunologyImmunotherapy and Immune Responsesvaccines and immunoinformatics approaches

Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers

Hadrup, Sine Reker, Bakker, Arnold H, Shu, Chengyi J, Andersen, Rikke S, van Veluw, Jerre, Hombrink, Pleun, Castermans, Emilie, thor Straten, Per +4 more·2009·Nature Methods

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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Crossref · 10.1038/nmeth.1345

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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 .”

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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
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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