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. 2025:2904:179-191.
doi: 10.1007/978-1-0716-4414-0_13.

Flow Cytometric Analysis of Mouse Brain CD8+ T Cell Activation

Du Huy Nguyen  1 , Manja Wibke Thiem  1 , Alexander Winston Veltkamp  1 , Ines Sophie Schädlich  1 , Jiani Jiang  1 , Mamta Panda  1 , Enrico Maria Costanzo  1 , Tim Magnus  1 , Björn Rissiek  2
Affiliations

Flow Cytometric Analysis of Mouse Brain CD8+ T Cell Activation

Du Huy Nguyen et al. Methods Mol Biol. 2025.

Abstract

Cytotoxic CD8+ T cell lymphocytes are pivotal for fighting infections with intracellular pathogens, e.g., viruses. Moreover, recent studies have elucidated the role of CD8+ T cells in various neuroinflammatory diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis (MS), and stroke. Therefore, the characterization of brain CD8+ T cells at the single-cell level is pivotal for understanding their state and function, e.g., in preclinical mouse models. Here, flow cytometry emerges as a versatile tool offering distinct techniques tailored to different facets of T cell activation. In this chapter, we provide protocols utilizing multicolor flow cytometry and cytokine secretion assays to analyze brain CD8+ T cell activation via the detection of surface lineage markers, extracellular and intracellular activation markers such as CD39, CD279, and granzyme B, or via measuring cytokine expression levels.

Keywords: CD279; CD39; Cytokine secretion; Cytotoxic CD8+ T cell lymphocytes; Granzyme B; Multicolor flow cytometry; Surface lineage markers.

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