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Phenotypic and Functional Characterization of Memory CD4+ and CD8+ T Cells After Antigenic Stimulation

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Immune Homeostasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2782))

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Abstract

The encounter of T cells with the antigen through the interaction of T cell receptors with peptides and major histocompatibility complex (MHC) molecules on the surface of antigen-presenting cells (APCs) can generate effector response and memory T cells. Memory T cells developed following infections or vaccination may persist, leading to the generation of a specific immune response upon reexposure to the same pathogen through rapid clonal proliferation and activation of effector functions. T cell memory subsets can be identified based on the expression of several membrane markers such as CCR7, CD27, and CD45RA. Using fluorescent antibodies against these markers and a flow cytometer, it is possible to perform immunophenotyping via the analysis of cell surface expression of proteins by different subpopulations such as the subsets of naïve, effector, and memory T cells as well as via the analysis of functional markers that further characterize each sample. Intracellular cytokine staining allows for the evaluation of intracellular proteins expressed in T cells in response to antigenic stimulation. This chapter presents the phenotypic and functional characterization of memory T cells after antigenic stimulation, detailing the procedures for identifying intracellular and surface protein markers. Herein, we review and present a reproducible standardized protocol using antibodies for specific markers and applying flow cytometry.

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Acknowledgements

We are thankful to Dr. Catherine Riou from the Institute of Infectious Disease and Molecular Medicine at the University of Cape Town, South Africa, for her invaluable assistance in teaching us the FACS analysis presented in Fig. 3.

Funding

This work was supported by the Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG), grant ID 202010267000284. Carolina do Prado Servian and Letícia Carrijo Masson received scholarships from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Simone GonÓalves Fonseca is a research fellow of the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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Author notes

  1. Carolina do Prado Servian and Letícia Carrijo Masson contributed equally with all other contributors.

Authors and Affiliations

  1. Department of Bioscience and Technology, Institute of Tropical Pathology and Public Health, Federal University of Goiás, Goiânia, Goiás, Brazil

    Carolina do Prado Servian, Letícia Carrijo Masson & Simone Gonçalves Fonseca

Authors
  1. Carolina do Prado Servian
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  2. Letícia Carrijo Masson
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  3. Simone Gonçalves Fonseca
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Corresponding author

Correspondence to Simone Gonçalves Fonseca .

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Editors and Affiliations

  1. Department of Surgery, Morehouse School of Medicine, Atlanta, GA, USA

    Vijay Kumar

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do Prado Servian, C., Masson, L.C., Fonseca, S.G. (2024). Phenotypic and Functional Characterization of Memory CD4+ and CD8+ T Cells After Antigenic Stimulation. In: Kumar, V. (eds) Immune Homeostasis. Methods in Molecular Biology, vol 2782. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3754-8_14

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  • DOI: https://doi.org/10.1007/978-1-0716-3754-8_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3753-1

  • Online ISBN: 978-1-0716-3754-8

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