Multiplexed Peptide-MHC Tetramer Staining with Mass Cytometry

Part of the Methods in Molecular Biology book series (MIMB, volume 1346)

Abstract

Mass cytometry is flow cytometry based on single cell mass spectrometry with decreased crosstalk between channels and an ability to probe >40 parameters per cell, making it well suited for multiplexed assays. Peptide major histocompatibility (MHC) tetramer staining allows direct detection of antigen specific cells and is also amenable to multiplexing/combinatorial approaches. Here we describe methods for multiplexed pMHC-tetramer staining using mass cytometry.

Key words

Mass cytometry Peptide-MHC tetramer Antigen-specific T cell Combinatorial tetramer staining 

References

  1. 1.
    Davis MM et al (1998) Ligand recognition by alpha beta T cell receptors. Annu Rev Immunol 16(9099):523–544CrossRefPubMedGoogle Scholar
  2. 2.
    Altman JD et al (1996) Phenotypic analysis of antigen-specific T lymphocytes. Science 274(5284):94–96CrossRefPubMedGoogle Scholar
  3. 3.
    Davis MM, Altman JD, Newell EW (2011) Interrogating the repertoire: broadening the scope of peptide-MHC multimer analysis. Nat Rev Immunol 11(8):551–558PubMedCentralCrossRefPubMedGoogle Scholar
  4. 4.
    Newell EW, Davis MM (2014) Beyond model antigens: high-dimensional methods for the analysis of antigen-specific T cells. Nat Biotechnol 32(2):149–157PubMedCentralCrossRefPubMedGoogle Scholar
  5. 5.
    Newell EW et al (2012) Cytometry by time-of-flight shows combinatorial cytokine expression and virus-specific cell niches within a continuum of CD8+ T cell phenotypes. Immunity 36(1):142–152PubMedCentralCrossRefPubMedGoogle Scholar
  6. 6.
    Newell EW, Lin W (2014) High-dimensional analysis of human CD8(+) T cell phenotype, function, and antigen specificity. Curr Top Microbiol Immunol 377:61–84PubMedGoogle Scholar
  7. 7.
    Bandura DR et al (2009) Mass cytometry: technique for real time single cell multitarget immunoassay based on inductively coupled plasma time-of-flight mass spectrometry. Anal Chem 18:6813–6822CrossRefGoogle Scholar
  8. 8.
    Bendall SC et al (2011) Single-cell mass cytometry of differential immune and drug responses across a human hematopoietic continuum. Science 332(6030):687–696PubMedCentralCrossRefPubMedGoogle Scholar
  9. 9.
    Bodenmiller B et al (2012) Multiplexed mass cytometry profiling of cellular states perturbed by small-molecule regulators. Nat Biotechnol 30(9):858–867PubMedCentralCrossRefPubMedGoogle Scholar
  10. 10.
    Akram A, Inman RD (2012) Immunodominance: a pivotal principle in host response to viral infections. Clin Immunol 143(2):99–115CrossRefPubMedGoogle Scholar
  11. 11.
    Newell EW et al (2009) Simultaneous detection of many T-cell specificities using combinatorial tetramer staining. Nat Methods 6(7):497–499PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Hadrup SR et al (2009) Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers. Nat Methods 6(7):520–526CrossRefPubMedGoogle Scholar
  13. 13.
    Newell EW et al (2013) Combinatorial tetramer staining and mass cytometry analysis facilitate T-cell epitope mapping and characterization. Nat Biotechnol 31(7):623–629PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Ramachandiran V et al (2007) A robust method for production of MHC tetramers with small molecule fluorophores. J Immunol Methods 319(1-2):13–20PubMedCentralCrossRefPubMedGoogle Scholar
  15. 15.
    Howarth M, Ting AY (2008) Imaging proteins in live mammalian cells with biotin ligase and monovalent streptavidin. Nat Protoc 3(3):534–545PubMedCentralCrossRefPubMedGoogle Scholar
  16. 16.
    Bakker AH et al (2008) Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7. Proc Natl Acad Sci U S A 105(10):3825–3830PubMedCentralCrossRefPubMedGoogle Scholar
  17. 17.
    Toebes M et al (2006) Design and use of conditional MHC class I ligands. Nat Med 12(2):246–251CrossRefPubMedGoogle Scholar
  18. 18.
    Fuss IJ et al (2009) Isolation of whole mononuclear cells from peripheral blood and cord blood. Curr Protoc Immunol Chapter 7:Unit7.1Google Scholar
  19. 19.
    Yokoyama WM, Thompson ML, Ehrhardt RO (2012) Cryopreservation and thawing of cells. Curr Protoc Immunol. Appendix 3:3GGoogle Scholar
  20. 20.
    Leipold MD, Maecker HT (2012) Mass cytometry: protocol for daily tuning and running cell samples on a CyTOF mass cytometer. J Vis Exp 69, e4398PubMedGoogle Scholar
  21. 21.
    Lissina A et al (2009) Protein kinase inhibitors substantially improve the physical detection of T-cells with peptide-MHC tetramers. J Immunol Methods 340(1):11–24PubMedCentralCrossRefPubMedGoogle Scholar
  22. 22.
    Fienberg HG et al (2012) A platinum-based covalent viability reagent for single-cell mass cytometry. Cytometry A 81(6):467–475CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.School of Biological SciencesNanyang Technological UniversitySingaporeSingapore
  2. 2.Singapore Immunology Network (SIgN), Agency for Science Technology and Research (A-STAR)SingaporeSingapore

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