Fig. 6. Genetic amplification of NLRP3 promotes HPD in response to anti–PD-1 immunotherapy.

(A) Incidence of NLRP3 amplification in human tumor types based on The Cancer Genome Atlas (TCGA). Data were visualized using cBioPortal. (B) Flow cytometry analysis of PMN-MDSCs in the lungs of Ctrl and NLRP3a tumor–bearing mice (n = 3). (C) Left: S100β IHC of lungs derived from Ctrl and NLRP3a tumor–bearing mice. 20×; scale bars, 50 μm. Red arrows, S100β-positive cells. Right: Quantification of S100β⁺ cells in the lungs of Ctrl and NLRP3a tumor–bearing mice (n = 3). (D) Left: Flow cytometry analysis of lung tissues resected from TLR4^+/+ control and SPC-TLR4^−/− mice harboring control BRAF^V600EPTEN^−/− tumors or BRAF^V600EPTEN^−/−-NLRP3a tumors (n = 3). Right: Wnt5a qrt-PCR analysis of lung tissues from TLR4^+/+ control and SPC-TLR4^−/− mice harboring control BRAF^V600EPTEN^−/− tumors or BRAF^V600EPTEN^−/−-NLRP3a tumors (n = 3). (E) ELISA analysis of plasma HSP70 concentrations in Ctrl and NLRP3a tumor–bearing mice after IgG Ctrl or anti–PD-1 treatment (n = 3). Statistical analysis was performed by two-way ANOVA followed by Tukey’s multiple comparisons test. (F) Flow cytometry analysis of PMN-MDSCs in the lungs of Ctrl and NLRP3a tumor–bearing mice after anti–PD-1 treatment (n = 3). (G) Ctrl and NLRP3a TGRs after IgG Ctrl or anti–PD-1 treatment. Tumor growth is expressed as a ratio of anti–PD-1 treatment to IgG Ctrl treatment. All two-group comparisons were analyzed using unpaired t tests. All data are representative of two to three independent experiments and expressed as mean values ± SEM. *P < 0.05 and ***P < 0.0005.