Extended Data Fig. 10: ECSIT deficiency inhibits fumarate production of CD8+ memory T cells in chronic stimulation, and DMF supplementation improves the antitumour function of ECSIT-deficient memory CD8+ T cells in vivo.
(a-d) Ecsitfl/fl OT-1 (WT) and Ecsitfl/fl Rosa26CreERT2 OT-1 (iKO) splenocytes were activated with OVA and IL-2 for 2 days, followed by sorting of CD8+ T cells, and then sorted CD8+ T cells were cultured in IL-15 and OVA for 3 days to mimic tumour antigen chronic stimulation in the presence of 4-OHT. CD8+ T cells were collected for metabolite detection. (a) Schematic of CD8+ T cells stimulated chronically in vitro. (b) PCA analysis of metabolomics data. (c) Pathway enrichment analysis of changed metabolites. (d) Heat map of relative abundance of significantly changed metabolites. (e) Schematic of tumour model with DMF supplement. An equal number of OT-1 naive CD8+ T cells from Ecsitfl/+ CD45.1.2 OT-1 and Ecsitfl/fl Rosa26CreERT2 CD45.2 OT-1 mice were mixed and transferred into CD45.1 recipient mice that had been engrafted with B16F10-OVA melanoma cells. Tamoxifen was administered every day for five treatments starting at day 11 to day 15 and DMF was also injected every other day from day 11 to day 20. OT-1 cells from the tumour were analysed on day 20. One-side Fisher’s exact test for (c).
