Extended Data Fig. 6: Elevated numbers and cytokine production by CD8+ T cells after immunotherapy.
From: Phosphoinositide acyl chain saturation drives CD8+ effector T cell signaling and function
a–d, Sex-matched C57BL/6 mice were injected in the right flank with 1 × 106 B16-F10-OVA cells. After 3 days, mice were given i.p. injections of anti-PD1 and anti-CTLA4 (aPD1 + aCTLA4) or matching isotype controls (IgG1 + IgG2) every 3 days for 3 rounds in total. Tumor growth was measured up to day 16 post-tumor injection and CD8+ T cells were isolated on day 12 post-tumor injection to measure T cell function and lipid composition. a, CD8+ tumor infiltrating lymphocytes (TILs) on day 12 post-tumor injection normalized to tumor weight. n = 7 IgG1 + IgG2 and n = 8 aPD1 + aCTLA4 biologically independent samples; unpaired two-tailed t-test. b–c, TILs were isolated day 12 post-tumor injection and restimulated with PMA/ionomycin plus brefeldin A for 5 h in vitro. n = 7 biologically independent samples per group, unpaired two-tailed t-test. b, Representative contour plots of IFN-γ expression (left panel), percentage IFN-γ+ (middle panel) and IFN-γ expression (right panel) are shown. c, Representative contour plots of Granzyme B expression (left panel), percentage Granzyme B+ (middle panel) and Granzyme B expression (right panel) are shown. d, Relative total PI calculated from quantile normalized lipid peak areas. n = 7 IgG1 + IgG2 and n = 8 aPD1 + aCTLA4 biologically independent samples; unpaired two-tailed t-test. e-f, Sex-matched C57BL/6 mice were injected in the right flank with 1x106 B16-F10-OVA cells. After 3 days, mice were given i.p. injections of anti-PD1 antibody or matching isotype control (IgG2) every 3 days for 3 rounds in total. Tumor growth was measured up to day 12 post-tumor injection and CD8+ T cells were isolated on day 12 post-tumor injection to measure T cell function and lipid composition. e, Average tumor diameter measured at the indicated time points. n = 5 biologically independent samples per group; two-way ANOVA corrected for multiple comparisons (Sidak test) comparing the two groups at each time point across the dataset. f, Lipids were analyzed from TILs on day 12 post-tumor injection. Saturation of acyl chains is expressed as a percentage of total PI. n = 5 biologically independent samples per group; unpaired two-tailed t-test comparing saturated PI. Error bars show the standard error of the mean.
