Protocol for the quantitative detection of mtDNA in the supernatants of activated human naive CD8+ T cells

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Here, we present a protocol for the quantitative characterization of human T cell aging. We describe steps for sample collection; peripheral blood mononuclear cell (PBMC) isolation; and the enrichment, assessment, and activation of naive CD8 + T cells. We then detail procedures for supernatant collection and quantification using the absolute copy number of mitochondrial DNA (mtDNA) released into the supernatants from in - vitro -activated naive CD8 + T cells. We also apply this approach to predict the occurrence of lung adenocarcinoma in middle-aged populations. For complete details on the use and execution of this protocol, please refer to Jin et al. 1 • Steps for the quantification of mtDNA release by activated naive CD8 + T cells in vitro • Optimization of the manipulating temperature for the isolation of naive CD8 + T cells • Utilization of mtDNA release by activated T cells to predict lung adenocarcinoma Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for the quantitative characterization of human T cell aging. We describe steps for sample collection; peripheral blood mononuclear cell (PBMC) isolation; and the enrichment, assessment, and activation of naive CD8 + T cells. We then detail procedures for supernatant collection and quantification using the absolute copy number of mitochondrial DNA (mtDNA) released into the supernatants from in - vitro -activated naive CD8 + T cells. We also apply this approach to predict the occurrence of lung adenocarcinoma in middle-aged populations.

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