Figure 5
Thermal ablation stimulates a weak adaptive response that is augmented by TLR ligation and checkpoint modulation. (A) Z-scores of adaptive immune and T-cell response related genes for each treatment cohort. (B) CIBERSORTx-imputed absolute immune cell content. (C) CIBERSORTx-imputed T-cell phenotype distribution. Flow cytometry quantitation of (D) CD8+ and (E) CD4+ T-cells in tumors treated with a combined immunotherapy-HIFU protocol. Myeloid derived suppressor cells (MDSCs) as quantified with flow cytometry of the (F) thermal ablation and (G) mechanical ablation cohorts. Pdcd1 IHC stained sections of (H) an untreated control tumor, (I) a thermally-ablated tumor, (J) a distant tumor in an animal that received thermal ablation, (K) a tumor that received both thermal ablation and immunotherapy, (L) a distant tumor in an animal that received both thermal ablation and immunotherapy, and (M) a tumor treated with mechanical HIFU. FoxP3 IHC stained sections of (N) an untreated control tumor, (O) a thermally-ablated tumor, (P) a distant tumor in an animal that received thermal ablation, (Q) a tumor that received both thermal ablation and immunotherapy, (R) a distant tumor in an animal that received both thermal ablation and immunotherapy, and (S) a tumor treated with mechanical HIFU. Scale bar represents 100 µm in all cases. Time points for RNA-seq and IHC data are: 1-week post ablation for all cohorts that received thermal ablation, and 24 h post HIFU for cohorts that received mechanical HIFU. For flow cytometry studies, time points are 72 h for cohorts that received mechanical HIFU or immunotherapy in addition to thermal ablation and 1 week for cohorts that received thermal ablation alone. Data are expressed as mean ± SD. *p < 0.05, **p < 0.01, ****p < 0.0001 (ordinary one-way ANOVA with Tukey correction for multiple hypotheses). The heatmap was created in R Studio v1.2.5001 (https://rstudio.com).
