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Exploration of T-Cell Diversity Using Mass Cytometry

  • Kaitlin C. O’Boyle
  • Takuya Ohtani
  • Sasikanth Manne
  • Bertram Bengsch
  • Sarah E. Henrickson
  • E. John WherryEmail author
  • Cecile AlanioEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2111)

Abstract

T-cell diversity is multifactorial and includes variability in antigen specificity, differentiation, function, and cell-trafficking potential. Spectral overlap limits the ability of traditional flow cytometry to fully capture the diversity of T-cell subsets and function. The development of mass cytometry permits deep immunoprofiling of T-cell subsets, activation state, and function simultaneously from even small volumes of blood. This chapter describes our methods for mass cytometry and high-throughput data analysis of T cells in patient cohorts. We provide a pipeline that includes practical considerations when customizing a panel for mass cytometry. We also provide protocols for the conjugation and titration of metal-labeled antibodies (including two T-cell panels) and a staining procedure. Finally, with the aim to support translational science, we provide R scripts that contain a detailed workflow for initial evaluation of high-dimensional data generated from cohorts of patients.

Key words

Mass cytometry CyTOF T cells Systems biology High-throughput analysis Clustering Data processing 

Notes

Acknowledgements

We thank Divij Matthew, University of Pennsylvania, USA, for his help with the titration figure, as well as Jacob Bergstedt, Lund University, Sweden, for his help with the R scripts.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Kaitlin C. O’Boyle
    • 1
  • Takuya Ohtani
    • 1
  • Sasikanth Manne
    • 1
  • Bertram Bengsch
    • 2
  • Sarah E. Henrickson
    • 1
    • 3
  • E. John Wherry
    • 1
    • 4
    Email author
  • Cecile Alanio
    • 1
    • 4
    Email author
  1. 1.Department of Systems Pharmacology and Translational Therapeutics, Institute for Immunology, Perelman School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Department of Medicine II, Gastroenterology, Hepatology, Endocrinology, and Infectious DiseasesUniversity Medical Center Freiburg and Signaling Research Centres BIOSS and CIBSS, University of FreiburgFreiburg im BreisgauGermany
  3. 3.Division of Allergy Immunology, Department of PediatricsChildren’s Hospital of PhiladelphiaPhiladelphiaUSA
  4. 4.Parker Institute of Cancer ImmunotherapyUniversity of PennsylvaniaPhiladelphiaUSA

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