Next-generation detection of antigen-responsive T cells using DNA barcode-labeled peptidemajor histocompatibility complex I multimers

Amalie Kai Bentzen, Andrea Marion Marquard, Rikke Birgitte Lyngaa, Sunil Kumar Saini, Malene Rask Andersen, Marco Donia, I. M. Svane, P. Thor Straten, Z. Szallasi, S. N. Jakobsen, A. C. Eklund, Sine Reker Hadrup

    Research output: Contribution to journalConference abstract in journalResearchpeer-review

    Abstract

    Identification of antigenic peptides recognized by T cells is important for understanding and treatingimmune related diseases. Current cytometry-based approaches are limited to simultaneous screening of T cell reactivity towards 10-100 distinct peptide specificities, which poorly match the large diversity of T cell recognition in humans. Consequently it has been impossible to comprehensively analyze T cell responsiveness in cancer, infectious and autoimmune diseases. We present and validate a novel technology that enables parallel detection of numerous different peptide-MHC responsive T cells in asingle sample using >1000 different peptide-MHC multimers labeled with individual DNA barcodes. After isolation of MHC multimer binding T cells their recognition are revealed by amplification and sequencing of the MHC multimer-associated DNA barcodes. The relative frequency of the sequenced DNA barcodes originating from a given peptide-MHC motif relates to the size of the antigenresponsive T cell population. We have demonstrated the use of large panels of >1000 DNA barcoded MHC multimers for detection of rare T cell populations of virus and cancer-restricted origin in various tissues and compared with combinatorial encoding of fluorescent-labeled MHC multimers. Finally, we have demonstrated that this technology can be applied for multiplex T cell detection both in limited biological samples, such as uncultured tumor material, and for simultaneous assessment of target recognition and functional capability of T cells. This technology enables true high-throughput detection of antigen-responsive T cells and will advance our understanding of immune recognition from model antigens to genomewide immune assessments on a personalized basis.
    Original languageEnglish
    JournalEuropean Journal of Immunology
    Volume46
    Issue numberSuppl. 1
    Pages (from-to)9-9
    ISSN0014-2980
    Publication statusPublished - 2016
    Event ICI 2016 International Congress of Immunology - Melbourne, Australia
    Duration: 21 Aug 201626 Aug 2016

    Conference

    Conference ICI 2016 International Congress of Immunology
    CountryAustralia
    CityMelbourne
    Period21/08/201626/08/2016

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