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. 2022 Mar 14;10(5):581–596. doi: 10.1158/2326-6066.CIR-21-0831

Figure 4.

Figure 4. WTX-124 treatment increases immune cell activation and infiltration of MC38 tumors. MC38 tumor cells were implanted and allowed to grow to an average volume of 100–150 mm3 before mice were randomized into treatment groups. Mice were dosed twice a week with WTX-124 (100 μg) or PBS. Tumors were collected 24 hours after the second dose and dissociated for further analysis. A and B, RNA from each tumor was isolated and subjected to immune profiling with the NanoString PanCancer Mouse Immune Profiling Panel. A, Heatmap of transcripts with statistically significant differences in expression between the two treatments. Transcripts were excluded from the heatmap if they had average normalized counts below 50. Each lane represents an individual animal. B, Volcano plot of transcripts differentially expressed between WTX-124 and vehicle-treated mice. C, Specific pathway scores for WTX-124 or vehicle-treated mice. P values are derived from a two-way ANOVA with multiple comparisons (***, P < 0.001; ****, P < 0.0001). D, Normalized gene counts from selected immune checkpoint genes. E, Flow cytometry analysis of TIL density of various immune populations, including fold change information between the vehicle-treated and WTX-124–treated groups. F, The ratio of total CD8+ T cells or tetramer-positive CD8+ T cells to Tregs within the TILs, including fold change information between the vehicle and WTX-124–treated groups. G and H, The frequency of tetramer-positive CD8+ T cells producing IFNγ after restimulation with PMA/Ionomycin. I, The frequency of polyfunctional tetramer-positive CD8+ T cells by examining coexpression of IFNγ, TNF, and granzyme B after PMA/Ionomycin restimulation. The frequency tumor infiltrating FoxP3+ Tregs producing IFNγ (J and K) or TNF (L and M) after PMA/Ionomycin restimulation. Unless otherwise stated, data are presented as the mean ± SD, and P values are derived from t tests (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001). CTLA-4, CTL-associated protein 4; LAG-3, lymphocyte-activation gene 3; Tet, tetramer; TIGIT, T-cell immunoglobulin and ITIM (immunoreceptor tyrosine-based inhibitory motif) domain; TIM-3, T-cell immunoglobulin and mucin domain-containing protein 3.

WTX-124 treatment increases immune cell activation and infiltration of MC38 tumors. MC38 tumor cells were implanted and allowed to grow to an average volume of 100–150 mm3 before mice were randomized into treatment groups. Mice were dosed twice a week with WTX-124 (100 μg) or PBS. Tumors were collected 24 hours after the second dose and dissociated for further analysis. A and B, RNA from each tumor was isolated and subjected to immune profiling with the NanoString PanCancer Mouse Immune Profiling Panel. A, Heatmap of transcripts with statistically significant differences in expression between the two treatments. Transcripts were excluded from the heatmap if they had average normalized counts below 50. Each lane represents an individual animal. B, Volcano plot of transcripts differentially expressed between WTX-124 and vehicle-treated mice. C, Specific pathway scores for WTX-124 or vehicle-treated mice. P values are derived from a two-way ANOVA with multiple comparisons (***, P < 0.001; ****, P < 0.0001). D, Normalized gene counts from selected immune checkpoint genes. E, Flow cytometry analysis of TIL density of various immune populations, including fold change information between the vehicle-treated and WTX-124–treated groups. F, The ratio of total CD8+ T cells or tetramer-positive CD8+ T cells to Tregs within the TILs, including fold change information between the vehicle and WTX-124–treated groups. G and H, The frequency of tetramer-positive CD8+ T cells producing IFNγ after restimulation with PMA/Ionomycin. I, The frequency of polyfunctional tetramer-positive CD8+ T cells by examining coexpression of IFNγ, TNF, and granzyme B after PMA/Ionomycin restimulation. The frequency tumor infiltrating FoxP3+ Tregs producing IFNγ (J and K) or TNF (L and M) after PMA/Ionomycin restimulation. Unless otherwise stated, data are presented as the mean ± SD, and P values are derived from t tests (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001). CTLA-4, CTL-associated protein 4; LAG-3, lymphocyte-activation gene 3; Tet, tetramer; TIGIT, T-cell immunoglobulin and ITIM (immunoreceptor tyrosine-based inhibitory motif) domain; TIM-3, T-cell immunoglobulin and mucin domain-containing protein 3.