Extended Data Fig. 4: TGR TE have altered glucose reallocation upon re-exposure to drive anabolic metabolism.
From: Metabolic conditioning of CD8+ effector T cells for adoptive cell therapy
WT CD8+ T cells isolated from spleens of C57BL/6 mice were activated with anti-CD3 (5 μg/mL), anti-CD28 (0.5 μg/mL), IL-2 (100 U/mL), expanded for a total of 72 h, and exposed to 10 mM (control) or 1 mM (TGR) glucose during the final 5 min cells were re-exposed to normal (U-12C) glucose or heavy labelled (U-13C) glucose. 20 h cultures were started at 1 × 106 (control) and 1.5 × 106 (TGR) cells per/ml to generate similar end concentrations for glucose pulse experiments. a, Table showing the results of a Kegg-pathway analysis of the significantly increased and decreased metabolite pools in TGR TE after normal (U-12C) glucose re-feeding when compared to re-fed control TE. b, Table shows the results of a Kegg pathway analysis of the metabolites that exhibited increased glucose-derived U-13C assimilation (as determined by X13 CMS software) after re-feeding TGR TE with heavy labelled (U-13C) glucose compared to re-fed control TE. Data are from 3 biological replicates, representative of 3 independent experiments. Kegg Pathways were ranked according to the number of metabolite hits in that pathway (from highest to lowest). Metabolites in the highest ranked pathways were further validated by targeted analysis. c, Polar metabolites were extracted and untargeted metabolomic analysis was performed using XCMS on cells re-exposed to normal (U-12C) glucose. Volcano plot depicts relative metabolite pools compared between control and TGR cells. Orange circle represents a pentose phosphate pathway metabolite Pentose-phosphate, significantly increased in glucose re-fed TGR TE. Statistical significance was calculated using a Welch’s t-test, comparing relative intensities of each isotopologue in labelled samples of control TE versus those of TGR TE.
