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Multiparametric Flow Cytometry Analysis of Naïve, Memory, and Effector T Cells

  • Ankit Saxena
  • Pradeep K. Dagur
  • Angélique BiancottoEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 2032)

Abstract

Polychromatic flow cytometry enables the detection and characterization of markers which are helpful in defining phenotype of various cell subsets. Here we describe flow cytometry-based method to characterize phenotype of naïve, memory, and effector T cells. Being able to differentiate these cells is crucial in understanding immune response, and immune profiling. Naïve T cells enable the body to fight off new, unrecognized infections and diseases, and memory T cells are enriched for response to recall antigens. Furthermore, the antigen-experienced T cell populations can be broadly divided into effector and memory cell compartments, both of which are needed for sustaining a responsive immune system. Simplistically, the effector T cells require active antigenic stimulation to eliminate pathogens. On the other hand, memory T cells are described as cells which remain present in the absence of antigenic stimulation and have the capacity to expand rapidly upon secondary challenges. Recently, with the identification of central and effector memory T cell subsets, tremendous efforts have been devoted to characterize markers on the surfaces of these cells. Though, various markers have been used to identify the subsets, no single marker that segregates one subset from the other has been described. Thus, multiple markers are needed to subset the cells in order to characterize them. Here we report the verification of a nine-color panel (CD3, CD4, CD8, CD45RO, CD28, CD95, CCR7, Live/Dead Aqua, dump channel-CD19, CD14, CD56, CD16) that can successfully identify six distinct CD4 and CD8 T cell populations within the naïve and effector cell subsets from human donors.

Key words

Immunophenotyping Multicolor flow cytometry Whole blood T cells Memory T cells Naïve T cells 

References

  1. 1.
    Maecker HT, McCoy JP, Nussenblatt R (2012) Standardizing immunophenotyping for the human immunology project. Nat Rev Immunol 12(3):191–200.  https://doi.org/10.1038/nri3158CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    De Rosa SC, Herzenberg LA, Herzenberg LA et al (2001) 11-color, 13-parameter flow cytometry: identification of human naive T cells by phenotype, function, and T-cell receptor diversity. Nat Med 7(2):245–248.  https://doi.org/10.1038/84701CrossRefPubMedGoogle Scholar
  3. 3.
    Picker LJ, Singh MK, Zdraveski Z et al (1995) Direct demonstration of cytokine synthesis heterogeneity among human memory/effector T cells by flow cytometry. Blood 86(4):1408–1419PubMedGoogle Scholar
  4. 4.
    Sprent J, Cho JH, Boyman O et al (2008) T cell homeostasis. Immunol Cell Biol 86(4):312–319.  https://doi.org/10.1038/icb.2008.12CrossRefPubMedGoogle Scholar
  5. 5.
    Sprent J, Tough DF (1994) Lymphocyte life-span and memory. Science 265(5177):1395–1400CrossRefGoogle Scholar
  6. 6.
    Zhang N, Bevan MJ (2011) CD8(+) T cells: foot soldiers of the immune system. Immunity 35(2):161–168.  https://doi.org/10.1016/j.immuni.2011.07.010CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Mahnke YD, Brodie TM, Sallusto F et al (2013) The who's who of T-cell differentiation: human memory T-cell subsets. Eur J Immunol 43(11):2797–2809.  https://doi.org/10.1002/eji.201343751CrossRefPubMedGoogle Scholar
  8. 8.
    Golubovskaya V, Wu L (2016) Different subsets of T cells, memory, effector functions, and CAR-T immunotherapy. Cancers (Basel) 8(3).  https://doi.org/10.3390/cancers8030036CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Ankit Saxena
    • 1
  • Pradeep K. Dagur
    • 1
  • Angélique Biancotto
    • 2
    Email author
  1. 1.Flow Cytometry Core Facility, National Heart, Lung, and Blood InstituteNational Institutes of HealthBethesdaUSA
  2. 2.Precision ImmunologyImmunology and Inflammation Research Therapeutic AreaCambridgeUSA

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