Figure 8.
Mettl8 inhibition synergizes with anti–PD-1 treatment. (A) Schematic diagram of the combined Mettl8 KO and anti–PD-1 treatment model: CD45.2 mice were subcutaneously injected with 2 × 105 EG7-OVA cells, followed by 5 × 105 CD45.1.2 WT or Mettl8−/− OT-I cells transfer at 9 dpi. Anti–PD-1 antibody was administered i.p. every 2 days from 11 to 17 dpi. Mice were harvested at 19 dpi. (B–D) Tumor growth (B), tumor weight (C), and absolute number of tumor infiltrating OT-I cells (D) of the mice in A. n = 6 per group. (E and F) Representative flow cytometry plots (E) and cumulative data (F) show the frequency and absolute number of Tcf1+ Tim3− TPEX and Tim3+ Tcf1− TEX cells gated on tumor-infiltrating OT-I cells. n = 6 per group. (G and H) Representative flow cytometry plots (G) and cumulative data (H) show the frequency and absolute number of CX3CR1+ Tcf1− Int-TEX cells gated on tumor-infiltrating OT-I cells. n = 6 per group. (I) Schematic diagram of the combined GA and anti–PD-1 treatment model: CD45.2 mice were subcutaneously injected with 2 × 105 EG7-OVA cells, followed by 2 × 106 CD45.1.2 OT-I cell transfer at 9 dpi. GA and anti–PD-1 antibody were administered i.p. in an alternating schedule from 11 to 19 dpi. Mice were harvested at 23 dpi. (J) Tumor growth (left) and survival curve (right) in each group of the mice in I. n = 8–9 per group. (K and L) Representative flow cytometry plots (K) and cumulative data (L) show the frequency and absolute number of Tcf1+ Tim3− TPEX and Tim3+ Tcf1− TEX cells gated on tumor-infiltrating OT-I cells. n = 6–7 per group. Data are representative of two independent experiments. P value was calculated by two-way ANOVA (B and J), two-tailed Student’s t test (C, D, F, H, and L), and Log-rank test (J); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Refer to the image caption for details. Panel A shows schematic diagram of O T-I adoptive transfer and anti-P D-one treatment timeline. Panel B shows a line graph of tumor growth across treatment groups. Panel C shows a scatter graph comparing tumor weight between groups. Panel D shows a scatter graph quantifying tumor-infiltrating O T-I cell numbers. Panel E shows representative flow cytometry plots of T c f one positive T i m three negative T P E X and T i m three positive T c f one negative T E X cells. Panel F shows scatter graphs quantifying frequencies and numbers of T P E X and T E X cells. These graphs compare subset distributions among W T-I g G, W T-anti-P D-one, K O-I g G, and K O-anti-P D-one groups. Panel G shows representative flow cytometry plots of C X three C R one positive T c f one intermediate T E X cells. Panel H shows scatter graphs quantifying frequency and absolute number of C X three C R one positive T c f one intermediate T E X cells. These graphs compare intermediate T E X populations across treatment conditions. Panel I shows schematic diagram of combined G A and anti-P D-one treatment schedule. Panel J shows a tumor growth line graph and a survival curve. These graphs compare therapeutic responses among P B S, DMSO, G A, and anti-P D-one combination treatments. Panel K shows representative flow cytometry plots of T c f one positive T i m three negative T P E X and T i m three positive T c f one negative T E X cells. Panel L shows scatter graphs quantifying frequencies and numbers of T P E X and T E X cells. These graphs evaluate T cell subset changes after combined G A and anti-P D-one treatment.

Mettl8 inhibition synergizes with anti–PD-1 treatment. (A) Schematic diagram of the combined Mettl8 KO and anti–PD-1 treatment model: CD45.2 mice were subcutaneously injected with 2 × 105 EG7-OVA cells, followed by 5 × 105 CD45.1.2 WT or Mettl8−/− OT-I cells transfer at 9 dpi. Anti–PD-1 antibody was administered i.p. every 2 days from 11 to 17 dpi. Mice were harvested at 19 dpi. (B–D) Tumor growth (B), tumor weight (C), and absolute number of tumor infiltrating OT-I cells (D) of the mice in A. n = 6 per group. (E and F) Representative flow cytometry plots (E) and cumulative data (F) show the frequency and absolute number of Tcf1+ Tim3− TPEX and Tim3+ Tcf1− TEX cells gated on tumor-infiltrating OT-I cells. n = 6 per group. (G and H) Representative flow cytometry plots (G) and cumulative data (H) show the frequency and absolute number of CX3CR1+ Tcf1− Int-TEX cells gated on tumor-infiltrating OT-I cells. n = 6 per group. (I) Schematic diagram of the combined GA and anti–PD-1 treatment model: CD45.2 mice were subcutaneously injected with 2 × 105 EG7-OVA cells, followed by 2 × 106 CD45.1.2 OT-I cell transfer at 9 dpi. GA and anti–PD-1 antibody were administered i.p. in an alternating schedule from 11 to 19 dpi. Mice were harvested at 23 dpi. (J) Tumor growth (left) and survival curve (right) in each group of the mice in I. n = 8–9 per group. (K and L) Representative flow cytometry plots (K) and cumulative data (L) show the frequency and absolute number of Tcf1+ Tim3− TPEX and Tim3+ Tcf1− TEX cells gated on tumor-infiltrating OT-I cells. n = 6–7 per group. Data are representative of two independent experiments. P value was calculated by two-way ANOVA (B and J), two-tailed Student’s t test (C, D, F, H, and L), and Log-rank test (J); *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal