Extended Data Fig. 8: BATF overexpression markedly enhances CD8+ T cell antitumour responses.
From: Targeting REGNASE-1 programs long-lived effector T cells for cancer therapy
a, OT-I cells were stimulated with anti-CD3 and anti-CD28 overnight before viral transduction, and then cultured in IL-7- and IL-15-containing medium for another 3 days in vitro. Control-sgRNA- and Regnase-1-sgRNA-transduced OT-I cells were then stimulated with anti-CD3, IL-2 or IL-21 overnight for flow cytometry analysis of BATF expression (top), and quantification of the MFI of BATF (bottom) (n = 6 samples per group). Numbers in graphs indicate the MFI (top) and fold change between comparisons (bottom). b–h, OT-I cells transduced with control retrovirus (RV; mCherry+) were mixed at a 1:1 ratio with cells transduced with Batf-overexpressing retrovirus (GFP+), and transferred into tumour-bearing hosts. Mice were analysed at day 4 (e) (n = 4 mice), day 5 (b, h) (n = 4 mice), day 7 (c, d, f, g) (n = 6–8 mice) or day 14 (c, d) (n = 6 mice) for the expression of BATF (b, left), active caspase-3 (f, left), IFNγ, GZMB, TNF and IL-2 (g, left) and TCF-1 (h, top) in TIL OT-I cells; the quantification of the MFI of BATF in TIL OT-I cells (b, right); quantification of the frequencies of active caspase-3+ cells (f, right), IFNγ+, GZMB+, TNF+ and IL-2+ cells (g, right) and TCF-1+ cells (h, bottom) in TIL OT-I cells; analysis of the proportion of donor-derived OT-I cells in total CD8α+ cells in TILs and spleen (c); the quantification of the relative OT-I cell percentage in CD8α+ cells in the spleen (normalized to input) (d); the dilution of CellTrace Violet (CTV) in TIL OT-I cells (e, left); and the quantification of the MFI of CTV in TIL OT-I cells (e, right). The numbers in graphs indicate the MFI (b, left; e, left), the frequencies of OT-I cells in gates (c), the frequency of active caspase-3+ cells (f, left), the frequencies of IFNγ+, GZMB+, TNF+ or IL-2+ cells (g, left), and the frequency of TCF-1+ cells (h, top). Mean ± s.e.m. (a, b, d–h). *P < 0.05, **P < 0.01, ***P < 0.001. Two-tailed unpaired Student’s t-test (a, b, d–h). Data are representative of two (a, c) independent experiments, or pooled from two (b, d–h) independent experiments.
