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

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Mass Cytometry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1989))

Abstract

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.

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Acknowledgments

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.

Author information

Authors and Affiliations

  1. Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN), Singapore, Singapore

    Yannick Simoni & Evan W. Newell

  2. Fred Hutch Cancer Research Center, Vaccine and Infectious Disease Division, Seattle, WA, USA

    Yannick Simoni & Evan W. Newell

  3. ImmunoSCAPE Pte. Ltd., Singapore, Singapore

    Michael Fehlings

Authors
  1. Yannick Simoni
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  2. Michael Fehlings
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  3. Evan W. Newell
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Corresponding author

Correspondence to Evan W. Newell .

Editor information

Editors and Affiliations

  1. Discipline of Pathology, The University of Sydney, Camperdown, NSW, Australia

    Helen M. McGuire

  2. Sydney Cytometry Facility, The University of Sydney and Centenary Institute, Camperdown, NSW, Australia

    Thomas M. Ashhurst

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Simoni, Y., Fehlings, M., Newell, E.W. (2019). Multiplex MHC Class I Tetramer Combined with Intranuclear Staining by Mass Cytometry. In: McGuire, H., Ashhurst, T. (eds) Mass Cytometry. Methods in Molecular Biology, vol 1989. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9454-0_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9454-0_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9453-3

  • Online ISBN: 978-1-4939-9454-0

  • eBook Packages: Springer Protocols

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