Extended Data Fig. 4: Metabolic profiling of IL-2-, H9- and H9T-expanded CD8+ T cells. | Nature

Extended Data Fig. 4: Metabolic profiling of IL-2-, H9- and H9T-expanded CD8+ T cells.

From: An engineered IL-2 partial agonist promotes CD8+ T cell stemness

Extended Data Fig. 4

a, Photograph showing the medium colour of mouse CD8+ T cells expanded for 8 days with IL-2, H9, or H9T. Data are representative of two independent experiments. b, c, CD8+ T cells were isolated and expanded for 8 days with IL-2, H9, or H9T, and 5 million cells were collected for metabolomics analysis; data correspond to this of Fig. 3a. Relative levels of glucose (b) and lactate (c) are presented as mean values ± SEM, one-way ANOVA test with Dunnett’s correction, n = 3 mice. Data are representative of two independent experiments. d, 8 day IL-2, H9, or H9T expanded CD8+ T cells were incubated with or without 2-NBDG to assess glucose uptake. Data are presented as mean values ± SEM, one-way ANOVA test with Dunnett’s correction, n = 4 mice. Data are representative of two independent experiments. e, Eight-day IL-2, H9, or H9T expanded CD8+ T cells were incubated with or without TMRM to assess mitochondrial membrane potential. Data are presented as mean values ± SEM, one-way ANOVA test with Dunnett’s correction, n = 8 mice. Data are representative of two independent experiments. fh, Pre-activated CD8+ T cells were treated with control or 1 mM 2-DG in the presence of IL-2 for 2 days. Cells were subsequently stained with antibodies to TCF-1 (n = 3 mice), CD62L (n = 3 mice), or pSTAT5 (n = 6 mice). Data are presented as mean values ± SEM, with two-tailed, paired t-test. Data are representative of two independent experiments. i, PCA plot of RNA-seq data from 2-DG and control treated cells. Pre-activated CD8+ T cells were treated with 10 nM IL-2 with or without 2-DG for 2 days, and RNA was extracted for library preparation. Data are from three mice

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