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Mass Cytometry Assays for Antigen-Specific T Cells Using CyTOF

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Flow Cytometry Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1678))

Abstract

T Cells specific for a single antigen tend to be rare, even after expansion of memory cells. They are commonly detected by in vitro stimulation with peptides or protein, followed by staining for intracellular cytokines. In this protocol, we use CyTOF® mass cytometry to collect single-cell data on a large number of cytokines/chemokines, as well as cell-surface proteins that characterize T cells and other immune cells. We also include a method for magnetic bead enrichment of antigen-stimulated T cells, based on their expression of CD154 and CD69. Coupling magnetic enrichment with highly multi-parameter mass cytometry, this method enables the ability to dissect the frequency, phenotype, and function of antigen-specific T cells in greater detail than previously possible. Rare cell subsets can be examined, while minimizing run times on the CyTOF.

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Acknowledgments

This work was supported by grants # 5U19AI057229 and S10RR027582 from the National Institutes of Health, and SFB 650 and the BCRT (FKZ 13GW0099).

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Authors and Affiliations

  1. Fluidigm Corporation, South San Francisco, CA, 94080, USA

    Dongxia Lin

  2. Human Immune Monitoring Center, Institute for Immunity, Transplantation, and Infection, Stanford University School of Medicine, Fairchild Science Building, 299 Campus Drive, Stanford, CA, 94305-5124, USA

    Holden T. Maecker

Authors
  1. Dongxia Lin
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  2. Holden T. Maecker
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Corresponding author

Correspondence to Holden T. Maecker .

Editor information

Editors and Affiliations

  1. Flow Cytometry Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Teresa S. Hawley

  2. Department of Anatomy and Regenerative Biology, School of Medicine and Health Sciences, George Washington University, Washington, District of Columbia, USA

    Robert G. Hawley

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Lin, D., Maecker, H.T. (2018). Mass Cytometry Assays for Antigen-Specific T Cells Using CyTOF. In: Hawley, T., Hawley, R. (eds) Flow Cytometry Protocols. Methods in Molecular Biology, vol 1678. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7346-0_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7346-0_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7344-6

  • Online ISBN: 978-1-4939-7346-0

  • eBook Packages: Springer Protocols

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