Real-time monitoring of mitochondrial respiration in cytokine-differentiated human primary T cells
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Real-time monitoring of mitochondrial respiration in cytokine-differentiated human primary T cells. / Mølgaard, Kasper; Rahbech, Anne; Met, Özcan; Svane, Inge Marie; Straten, Perthor; Desler, Claus; Peeters, Marlies J.W.
In: Journal of Visualized Experiments, Vol. 176, e62984, 2021.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Real-time monitoring of mitochondrial respiration in cytokine-differentiated human primary T cells
AU - Mølgaard, Kasper
AU - Rahbech, Anne
AU - Met, Özcan
AU - Svane, Inge Marie
AU - Straten, Perthor
AU - Desler, Claus
AU - Peeters, Marlies J.W.
N1 - Publisher Copyright: © 2021 JoVE Journal of Visualized Experiments.
PY - 2021
Y1 - 2021
N2 - During activation, the metabolism of T cells adapts to changes that impact their fate. An increase in mitochondrial oxidative phosphorylation is indispensable for T cell activation, and the survival of memory T cells is dependent on mitochondrial remodeling. Consequently, this affects the long-term clinical outcome of cancer immunotherapies. Changes in T cell quality are often studied by flow cytometry using well-known surface markers and not directly by their metabolic state. This is an optimized protocol for measuring real-time mitochondrial respiration of primary human T cells using an Extracellular Flux Analyzer and the cytokines IL-2 and IL-15, which differently affect T cell metabolism. It is shown that the metabolic state of T cells can clearly be distinguished by measuring the oxygen consumption when inhibiting key complexes in the metabolic pathway and that the accuracy of these measurements is highly dependent on optimal inhibitor concentration and inhibitor injection strategy. This standardized protocol will help implement mitochondrial respiration as a standard for T cell fitness in monitoring and studying cancer immunotherapies.
AB - During activation, the metabolism of T cells adapts to changes that impact their fate. An increase in mitochondrial oxidative phosphorylation is indispensable for T cell activation, and the survival of memory T cells is dependent on mitochondrial remodeling. Consequently, this affects the long-term clinical outcome of cancer immunotherapies. Changes in T cell quality are often studied by flow cytometry using well-known surface markers and not directly by their metabolic state. This is an optimized protocol for measuring real-time mitochondrial respiration of primary human T cells using an Extracellular Flux Analyzer and the cytokines IL-2 and IL-15, which differently affect T cell metabolism. It is shown that the metabolic state of T cells can clearly be distinguished by measuring the oxygen consumption when inhibiting key complexes in the metabolic pathway and that the accuracy of these measurements is highly dependent on optimal inhibitor concentration and inhibitor injection strategy. This standardized protocol will help implement mitochondrial respiration as a standard for T cell fitness in monitoring and studying cancer immunotherapies.
U2 - 10.3791/62984
DO - 10.3791/62984
M3 - Journal article
C2 - 34747403
AN - SCOPUS:85121984841
VL - 176
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
SN - 1940-087X
M1 - e62984
ER -
ID: 289391288