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

Rikke Sick Andersen, Pia Kvistborg, Thomas Mørch Frøsig, Natasja Wulff Pedersen, Rikke Birgitte Lyngaa, Arnold H. Bakker, Chengyi Jenny Shu, Per Thor Straten, Ton N. Schumacher, Sine Reker Hadrup

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Fluorescently labeled multimeric complexes of peptide-MHC, the molecular entities recognized by the T cell receptor, have become essential reagents for detection of antigen-specific CD8+ T cells by flow cytometry. Here we present a method for high-throughput parallel detection of antigen-specific T cells by combinatorial encoding of MHC multimers. Peptide-MHC complexes are produced by UV-mediated MHC peptide exchange and multimerized in the form of streptavidin-fluorochrome conjugates. Eight different fluorochromes are used for the generation of MHC multimers and, by a two-dimensional combinatorial matrix, these eight fluorochromes are combined to generate 28 unique two-color codes. By the use of combinatorial encoding, a large number of different T cell populations can be detected in a single sample. The method can be used for T cell epitope mapping, and also for the monitoring of CD8+ immune responses during cancer and infectious disease or after immunotherapy. One panel of 28 combinatorially encoded MHC multimers can be prepared in 4 h. Staining and detection takes a further 3 h.
Original languageEnglish
JournalNature Protocols (Print)
Volume7
Issue number5
Pages (from-to)891-902
Number of pages12
ISSN1754-2189
DOIs
Publication statusPublished - 2012
Externally publishedYes

Keywords

  • Biochemistry, Genetics and Molecular Biology (all)
  • antigen
  • epitope
  • fluorescent dye
  • H2 antigen
  • HLA antigen
  • quantum dot
  • animal
  • article
  • CD8+ T lymphocyte
  • chemistry
  • epitope mapping
  • fluorescence imaging
  • human
  • immunology
  • metabolism
  • methodology
  • mouse
  • Animals
  • Antigens
  • CD8-Positive T-Lymphocytes
  • Epitope Mapping
  • Epitopes, T-Lymphocyte
  • Fluorescent Dyes
  • H-2 Antigens
  • HLA Antigens
  • Humans
  • Mice
  • Optical Imaging
  • Quantum Dots
  • BIOCHEMICAL
  • CLASS-I LIGANDS
  • P/MHC MICROARRAYS
  • EPITOPES
  • TECHNOLOGY
  • DISCOVERY
  • CYTOMETRY
  • TETRAMERS
  • EXCHANGE

Fingerprint Dive into the research topics of 'Parallel detection of antigen-specific T cell responses by combinatorial encoding of MHC multimers'. Together they form a unique fingerprint.

Cite this