Mass Cytometry pp 147-158 | Cite as

Multiplex MHC Class I Tetramer Combined with Intranuclear Staining by Mass Cytometry

  • Yannick Simoni
  • Michael Fehlings
  • Evan W. NewellEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1989)


Antigen-specific CD8+ T cells play a crucial role in the host protective immune response against viruses, tumors, and other diseases. Major histocompatibility complex (MHC) class I tetramers allow for a direct detection of such antigen-specific CD8+ T cells. Using mass cytometry together with multiplex MHC class I tetramer staining, we are able to screen more than 1000 different antigen candidates simultaneously across tissues in health and disease, while retaining the possibility to deliver an in-depth characterization of antigen-specific CD8+ T cells and associated phenotypes. Here we describe the method for a MHC class I tetramer multiplexing approach together with intracellular antibody staining for a parallel phenotypic cell characterization using mass cytometry in human specimens.

Key words

Mass cytometry MHC class I tetramer Antigen-specific T cell Multiplex tetramer staining 



The authors thank all members of Evan Newell lab.

Competing interests: E.W.N. is a board director and shareholder of immunoSCAPE Pte.Ltd. M.F. is Director, Scientific Affairs and shareholder of immunoSCAPE Pte. Ltd. All other authors declare no competing financial interests.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yannick Simoni
    • 1
    • 2
  • Michael Fehlings
    • 3
  • Evan W. Newell
    • 1
    • 2
    Email author
  1. 1.Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN)SingaporeSingapore
  2. 2.Fred Hutch Cancer Research Center, Vaccine and Infectious Disease DivisionSeattleUSA
  3. 3.ImmunoSCAPE Pte. Ltd.SingaporeSingapore

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