Extended Data Fig. 1: RC is a unique metabolic feature of rapidly proliferating T_E cells.

a, Experimental set-up of expanding effector (T_E, blue) and resting memory (T_M, red) T cell differentiation. OVA-specific CD8⁺ T cells from OT1 mice were activated with SIINFEKL (OVA) peptide and cultured with IL-2 for 3 days and with IL-15 for 4 additional days. T_E cells were collected at day 3 and T_M cells were collected at day 7 post-activation. Before collection, cells were cultured with [U-¹³C]-glutamine for 2 h and labelling patterns were analysed by mass spectrometry. b, Bar graphs representing the percentages of citrate isotopologues m + 0, m + 1, m + 2, m + 3, m + 4, m + 5 and m + 6 detected by [U-¹³C]-glutamine labelling in T_E or T_M cells. (n = 4 biological replicates per group). c–e, Percentages of aspartate (c), malate (d) and fumarate (e) isotopologues m + 0, m + 1, m + 2, m + 3 and m + 4 detected by [U-¹³C]-glutamine labelling in T_E or T_M cells. f–h, Ratios of m + 3 over m + 4 isotopologues of aspartate (f), malate (g) and fumarate (h) detected by [U-¹³C]-glutamine labelling in T_E and T_M cells. (n = 4 biological replicates per group). i–k, Percentage of isotopologues of palmitate (i), myristate (j) and stearate (k) labelling from [U-¹³C]-glutamine in T_E or T_M cells. (n = 3 biological replicates per group (T_M cells) and n = 2 biological replicates per group (T_E cells)). l, Glutamine consumption in mouse CD8⁺ T_E and T_M cells over 24 h at day 3 and day 7 post-activation respectively. (n = 3 biological replicates per group). m, Percentages of KI67⁺ out of CD8⁺ T cells in T_E and T_M cells. (n = 6 biological replicates per group, pooled data from 2 independent experiments). n, Experimental set-up of yellow fever vaccine administration to healthy volunteers, from which yellow fever tetramer⁺ CD8⁺ T cells were isolated from the blood at indicated time points. o, Gene expression in CD8⁺ T cells from healthy human volunteers vaccinated with yellow fever vaccine. Shown here are IDH3A, B and G reads per kilobase of transcript per million reads mapped in indicated cell subsets isolated from peripheral blood. (n = 3 (T_E), n = 5 (T_M) and n = 6 (T_N) biological replicates). p, mRNA expression of Idh3a in indicated T cell subsets isolated from mouse spleens at day 7 or day 28 post-LM-OVA infection. Data are represented as fold change as compared to SLECs, normalized to β2-microglobulin expression. (n = 3 biological replicates). q, mRNA expression of Idh2 in indicated T cell subsets isolated from spleens of mice at day 7 or day 28 post-LM-OVA infection. Data are represented as fold change as compared to SLECs, normalized to β2-microglobulin expression. (n = 4 biological replicates). r, Representative immunoblot for IDH2 in control Scr or IDH2 deleted cells. s–v, Percentage of m + 1 cis-aconitate (s), fumarate (t), malate (u) and succinate (v) labelling from [1-¹³C]-glutamine in Scr or IDH2 deleted CD8⁺ T cells. (n = 3 biological replicates per group). Data represent mean ± s.e.m. and were analysed by unpaired, two-tailed Student’s t-test (c–h,l,m,s–v), multiple unpaired, two-tailed t-test (b,i–k) or one-way ANOVA using Tukey’s multiple comparison test (o–q). Only relevant statistical comparisons are shown. For gel source data, see Supplementary Fig. 1.

Source data