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

Amalie Kai Bentzen, Andrea Marion Marquard, Rikke Birgitte Lyngaa, Sunil Kumar Saini, S. R. Andersen, M. Donia, I. M. Svane, P. thor Straten, Zoltan Imre Szallasi, S. N. Jakobsen, Aron Charles 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 screeningof T cell reactivity towards 10-100 distinct peptide specificities, which poorly match the large diversityof T cell recognition in humans. Consequently it has been impossible to comprehensively analyze Tcell responsiveness in cancer, infectious and autoimmune diseases. We present and validate a noveltechnology 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 andsequencing of the MHC multimer-associated DNA barcodes. The relative frequency of the sequencedDNA barcodes originating from a given peptide-MHC motif relates to the size of the antigenresponsiveT cell population. We have demonstrated the use of large panels of >1000 DNA barcodedMHC multimers for detection of rareT cell populations of virus and cancer-restricted origin in various tissues and compared withcombinatorial encoding of fluorescent-labeled MHC multimers. Finally, we have demonstrated that thistechnology can be applied for multiplex T cell detection both in limited biological samples, such asuncultured tumor material, and for simultaneous assessment of target recognition and functionalcapability of T cells. This technology enables true high-throughput detection of antigen-responsiveT cells and will advance our understanding of immune recognition from model antigens to genomewideimmune assessments on a personalized basis.
    Original languageEnglish
    Article number1831
    JournalEuropean Journal of Immunology
    Volume46
    Issue numberS1
    Pages (from-to)8-8
    Number of pages1
    ISSN0014-2980
    DOIs
    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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