Journal: Advanced Science
Article Title: DADA Enhances CD8 + T Cell Stemness to Improve Anti‐Tumor Immunity and Immunotherapy Efficacy
doi: 10.1002/advs.202519765
Figure Lengend Snippet: DADA improves the efficacy of ACT and ICB immunotherapies. (A) Schematic experimental procedure in (B–F): CTV‐labeled OT‐1 cells, which had undergone an initial activation with OVA peptide for 2 days and subsequent stimulation with anti‐CD3/CD28 mAbs and IL‐2 in the presence of DADA for 4 days, were transferred into MC38‐OVA tumor‐bearing mice. (B–D) Tumor growth curves (B), images of tumors (C), and tumor weights (D) at 15 days after MC38‐OVA inoculation; n = 6. (E,F) Quantification of transferred OT‐1 CD8 + T (E), Ki67 + CD8 + T, PD‐1 + TIM‐3 + CD8 + T, TCF1 − TIM‐3 + CD8 + T, TCF1 + TIM‐3 − CD8 + T, IFN‐γ + CD8 + T, TNF‐α + CD8 + T, and GZMB + CD8 + T (F) cells from tumor; n = 6. (G) Schematic experimental procedure in (H–L): WT mice were fed with DADA‐containing or normal water from day ‐14 until day 0, and were injected (s.c.) with MC38 cells on day 0. Anti‐PD‐1 mAb were injected (i.p.) on day 9, 12 and 15. (H–J) Tumor growth curves (H), images of tumors (I), and tumor weights (J) at 15 days after MC38 inoculation; n = 7. The experiment was repeated three times. (K,L) Quantification of CD8 + T (K), Ki67 + CD8 + T, PD‐1 + TIM‐3 + CD8 + T, TCF1 − TIM‐3 + CD8 + T, TCF1 + TIM‐3 − CD8 + T, IFN‐γ + CD8 + T, TNF‐α + CD8 + T, and GZMB + CD8 + T (L) cells from tumor; n = 7. The experiment was repeated three times. (M) Schematic experimental procedure in (N–R): WT mice were fed with DADA‐containing or normal water from day ‐14 until day 0, and were injected (s.c.) with B16‐F10 cells on day 0. Anti‐PD‐1 mAb were injected (i.p.) on day 9, 12 and 15. (N‐P) Tumor growth curves (N), images of tumors (O), and tumor weights (P) at 17 days after B16‐F10 inoculation; n = 7. The experiment was repeated three times. (Q,R) Quantification of CD8 + T (Q), Ki67 + CD8 + T, PD‐1 + TIM‐3 + CD8 + T, TCF1 − TIM‐3 + CD8 + T, TCF1 + TIM‐3 − CD8 + T, IFN‐γ + CD8 + T, and TNF‐α + CD8 + T (R)cells from tumor; n = 7. The experiment was repeated three times. Data are presented as mean ± SD and are analyzed by two‐way ANOVA (B, H, and N), unpaired t test (D–F), and one‐way ANOVA (J–L, and P‐R); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns, not significant.
Article Snippet: For isolation of mouse CD8 + T cells, cells were purified from splenic lymphocytes using through magnetic activated cell sorting (MACS) using the Mouse CD8 + T Lymphocyte Negative Selection Kit (Cat#130‐104‐075, Miltenyi Biotec, USA).
Techniques: Labeling, Activation Assay, Injection
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![DADA inhibits tumor growth by potentiating <t>CD8</t> + T cell anti‐tumor immune responses. (A) Schematic experimental procedure in (B–E): WT mice were fed with DADA‐containing or normal water from day ‐14 until the experimental endpoints or until day 0, and were injected (subcutaneously [s.c.]) with 16‐F10 cells on day 0. (B,C) Tumor growth curves (B) and tumor weights (C) at 17 days after B16‐F10 inoculation; n = 6. The experiment was repeated three times. (D,E) Representative flow cytometry plots and quantification of CD8 + T (D), IFN‐γ + CD8 + T and TNF‐α + CD8 + T (E) cells from tumor; n = 6. The experiment was repeated twice. (F) Schematic experimental procedure for (G–J): WT mice were fed with DADA‐containing or normal water from day ‐14 until the experimental endpoints or until day 0, and were injected (s.c.) with MC38 cells on day 0. (G and H) Tumor growth curves (G) and tumor weights (H) at 17 days after MC38 inoculation; n = 6. The experiment was repeated three times. (I,J) Representative flow cytometry plots and quantification of CD8 + T (I), IFN‐γ + CD8 + T, TNF‐α + CD8 + T and GZMB + CD8 + T (J) cells from tumor; n = 6. The experiment was repeated twice. (K) Schematic experimental procedure for (K–O): WT mice were fed with DADA‐containing or normal water from day ‐14 until the experimental endpoints or until day 0, and were injected (intravenously [i.v.]) with B16‐F10 cells on day 0. (L,M) Appearance of lungs (L) and the number of tumor nodules (M) at 17 days after B16‐F10 inoculation; n = 7. The experiment was repeated twice. (N and O) Representative flow cytometry plots and quantification of CD8 + T (N), IFN‐γ + CD8 + T, TNF‐α + CD8 + T and GZMB + CD8 + T (O) cells from the lung; n = 7. The experiment was repeated twice. (P) Schematic experimental procedure for (Q–S): WT mice were fed with DADA‐containing or normal water for 14 days and were injected (intraperitoneally [i.p.]) with 100 µg of anti‐CD8α antibody weekly. The mice were injected (s.c.) with MC38 cells on day 0. (Q–S) Tumor growth curves (Q), tumor weights (R), and representative flow cytometry plots of CD8 + T cells (S) at 17 days after MC38 inoculation; n = 7. The experiment was repeated three times. Data are presented as mean ± SD and are analyzed by two‐way ANOVA (B, G, and Q) and one‐way ANOVA (C–E, H–J, M–O, and R); * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. ns, not significant.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_9152/pmc13159152/pmc13159152__ADVS-13-e19765-g002.jpg)
