Fig 2: 6PGD-/- CD8+ T cells display a TEFF phenotype(A) CD8+ T cells were isolated from spleen of 6PGD-/- and 6PGDfl/fl mice and stimulated for 48 h with aCD3 + aCD28 mAbs, followed by RNA extraction and RNA-sequencing analysis. Heatmap of gene expression for 6PGD-/- versus 6PGDfl/fl CD8+ T cells is shown.(B) For confirmation, naive (CD62Lhigh CD44low) CD8+ T cells were isolated and stimulated for 4 days with aCD3 + aCD28 mAbs and IL-2 (20 IU/mL). Expression of representative genes was confirmed by real-time PCR on day 0 and day 4.(C–M) Naive CD8+ T cells were stimulated for 4 days with aCD3 + aCD28 mAbs and IL-2 (20 IU/mL) followed by assessment of IFN-? expression by intracellular staining (C) and quantification of IFN-? production by ELISA (D). Expression of granzyme B (E–F), T-bet (G and H), Fas (CD98) (I and J), FasL (CD178) (K and L), and CD69 (M) was assessed by flow cytometry. Results are representative of three independent experiments with n = 6 mice per group. Error bars represent ± SEM. ***p < 0.001; ****p < 0.0001.

Fig 3: Attenuated CAF-induced antigen-mediated AICD effects of CD8+ T cells by Cal/ICG@MPs.a Scheme of attenuated myofibroblast-induced antigen-mediated AICD effect of CD8+ T cells by Cal/ICG@MPs. The cell supernatants were collected after tumor cells were treated with PBS, ICG, Cal/ICG, ICG@MPs, Cal@MPs, or Cal/ICG@MPs at the ICG concentration of 4 µg mL-1 and Cal concentration of 60 ng mL-1 for 4 h in the presence or absence of 808 nm laser irradiation (1 W cm-2, 5 min). Myofibroblasts were pretreated with PBS, ICG, Cal/ICG, ICG@MPs, Cal@MPs, or Cal/ICG@MPs at the ICG concentration of 4 µg mL-1 and Cal concentration of 60 ng mL-1 for 48 h, and then co-cultured with the above tumor cell supernatants for 24 h. Finally, the treated myofibroblasts were co-cultured with CD8+ T cells for another 48 h. The phenotype of myofibroblasts and CD8+ T cells was analyzed by flow cytometry. b MHC-I MFI in myofibroblasts after the pretreated myofibroblasts were co-cultured with the cell supernatants from the treated H22 cells indicated in a. Data are presented as means ± s.d. (n = 3 biologically independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). c, d FasL (c) and PD-L2 (d) expression in myofibroblasts after the pretreated myofibroblasts were co-cultured with the cell supernatants from the treated H22 cells, and then CD8+ T cells indicated in a. Data are presented as means ± s.d. (n = 3 biologically independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). e, f Proliferation (e) and apoptosis (f) ratios of CD8+ T cells after treatment are indicated in a. Data are presented as means ± s.d. (n = 3 biologically independent samples; one-way ANOVA followed by Tukey’s HSD post hoc test). g–i MHC-I (g), FasL (h), and PD-L2 (i) expression in CAFs of stroma-rich H22 tumor-bearing mice at 14 days after intravenous injection of PBS, ICG, Cal/ICG, ICG@MPs, Cal@MPs, or Cal/ICG@MPs at the ICG dosage of 8 mg kg-1 and Cal dosage of 120 µg kg-1 twice every 2 days, followed with or without 808 nm laser irradiation (1.5 W cm-2, 10 min) at 2 h after the last injection by flow cytometry. Data are presented as means ± s.d. (n = 6 mice per group; one-way ANOVA followed by Tukey’s HSD post hoc test). j, k The numbers (j) and ratios (k) of CD8+ T cells in tumor tissues of stroma-rich H22 tumor-bearing mice after treatment are indicated in g. Data are presented as means ± s.d. (n = 6 mice per group; one-way ANOVA followed by Tukey’s HSD post hoc test). Source data are provided as a Source Data file.

Fig 4: Anti-IL18 Mitigates Liver Injury in Intestinal Il17RA-Deficient MiceLittermate Il17rafl/fl and Il17rafl/fl × villin cre+ mice were pre-treated with anti-IL-18 (0.5 mg/mouse) 1 day prior to IV concanavalin A (Con A) (25 mg/kg) and sacrificed at 8 h post Con A (n = 7–9 mice/group).(A) Serum alanine aminotransferase (ALT).(B) Quantification of TUNEL staining.(C) Representative images of TUNEL-stained liver histology.(D and E) Liver Ifng (D) and Fasl (E) gene expression as measured by qRT-PCR. (A–B, D–E) Data are represented as mean + SEM (one-way ANOVA with multiple comparisons test). p<0.05*, <0.01**, <0.001***, <0.0001****

Fig 5: Activated PI3Kd T cells show increased cell death, FasL, and pFoxO1(A) Annexin-V+ T cells from healthy controls and patients with APDS stimulated with anti-CD3 plus anti-CD28 (n = 4).(B) Annexin-V staining of sorted naive (CD62LhiCD44lo) CD8+ cells stimulated with anti-CD3 plus anti-CD28 (n = 3).(C and D) OT-1 cells were transferred into CD45 congenic hosts subsequently infected with X31-OVA influenza. (C) Experimental outline. (D) Left: live OT-1 cell numbers. Right: Annexin-V+ cells, (n = 2, 2–3 mice/group).(E–G) OT-1 cells stimulated with OVA257–264 with or without CAL-101 or Z-VAD (n = 3–5). (E) Cell death. (F) Fasl mRNA. (G) Surface FasL Mean fluorescence intensity (MFI) (left) and a representative histogram at 48 h (right).(H) Viability of OT-1 cells stimulation with or without blocking FasL, 48 h (n = 4).(I) Top: T cells stimulated with anti-CD3 plus anti-CD28. Immunoblot for pAKTS473, AKT, pFoxO1T24/FoxO3aT32, and FoxO1. Bottom: T cells stimulated with anti-CD3. Immunoblot for pZAP70Y319/Y352, ZAP70, pERKT202/Y204, and ERK (representative blots, n = 3).(J) pFoxO1S256 of OT-1 cells stimulated with OVA257–264 (n = 3, representative histogram, with MFI indicated).(K) Surface FasL on viable GFP+ OT-1 cells retrovirally transduced with Migr (control), Migr-FoxO1, or Migr-FoxO1AAA (n = 3, representative histogram).Graphs show mean ± SEM. *p < 0.05. See Figure S1.