Extended Data Fig. 1: Inosine can support human T effector cell proliferation and function in the absence of glucose.
From: Inosine is an alternative carbon source for CD8+-T-cell function under glucose restriction
a, Metabolite array from the PM-M1 and PM-M2 Microplates (BioLog Inc). PBMC cells were stimulated with plate-bound anti-CD3 antibody for at least three days to generate active human T cells. Cells were then collected and incubated with various energy sources in a PM-M1 or PM-M2 microplate for 24 h, followed by Biolog redox dye mix MB incubation and measured spectrophotometrically at 590 nm. b, c, Naive CD8+ T cells from human PBMC were activated by plate-bound anti-CD3 and anti-CD28 antibodies in the indicated conditional media for 5 days. Cell proliferation and cell death of active CD8+ T cells were determined by CFSE dilution and 7AAD uptake, respectively. Data are presented as mean ± SD (n=4). ***, P=0.0007 for Glucose free versus Inosine. d, GD2-CAR T cells are generated using complete media, then switched to indicated conditional media and cultured for 48-72 h. LAN-1 cells were co-cultured with GD2-CAR T cells in the indicated conditional media, and cell death of LAN-1 was determined by calcein release with the Spectramax M2 microplate reader. Data are presented as mean ± SD (n=4). ***, P=5E-8 and 0.000006 for E:T ratio 10:1 and 5:1 for Glucose free versus Inosine respectively. e, The indicated proteins in GD2-CAR T cells were quantified by intracellular staining following PMA and ionomycin stimulation. Data are presented as mean ± SD (n=4). ***, P=0.0002, 0.0002, 0.0004 for Granzyme B, TNF-α, and IFN-γ for Glucose free versus Inosine, respectively. Data were analyzed by unpaired two-sided t-test (c–e). Sample size (n) represents biologically independent samples (c–e).
