Fig 2: Expression of Inflammasome Components in Human PBMCs Treated with LPS or MPLA.(A) Gene expression of IL1B and IL18 in human PBMCs 30 minutes after LPS or MPLA treatment. (B) Levels of cleaved IL-1ß and IL-18 produced by human PBMCs 6 and 24 hours after LPS or MPLA treatment. (C) Expression of NLRP3 (110 kDa) and precursors of Caspase-1 (45 kDa), IL-1B (31 kDa), and IL-18 (24 kDa) in human PBMCs 6 hours after LPS or MPLA treatments. Gels have been cropped for clarity; the bands were confirmed by the comparison with full-length gel images and molecular weight (Supplementary Figure S8). (D) Densitometry analysis of the Western Blot results. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, NS = not significant; n = 4 in each group, and data are representatives of three independent experiments.

Fig 3: Simvastatin improves clinical manifestations and inhibits pyroptosis in the dermis. (A) Patient’s clinical symptoms. (B) Hematoxylin & eosin (H&E) staining showing foam cells (black arrow) and Touton giant cells (white arrow). Scale bar, 200 µm. (C) Immunohistochemical staining for diagnosis. Scale bar, 100 µm. (D) Patient’s clinical symptoms before and after simvastatin treatment. (E) H&E staining showing foam cells and inflammatory cells treated with placebo (left side of body) and simvastatin (right side of body). Scale bar, 100 µm. (F) Immunofluorescence staining to detect HMGCR, NLRP3, ASC, caspase-1, GSDMD, IL-1, and IL-18 levels in lesions treated with the placebo (left side of body) and simvastatin (right side of body). Arrow (foam cells). Scale bar, 25 µm. ***p < 0.001, versus the placebo.

Fig 4: Exhausted CD4+ T cells revealed the increased expression of genes related to cell cycle, angiogenesis, and cytokines. During the process of exhaustion, restimulated CD4+ T cells exhibited altered expression of (A) cell cycle inhibitor p21, which was strongly elevated in CD4+ T cells cocultured with cancer cells, (B) upregulation of angiogenesis-related genes, and (C) overexpression of interleukins IL-6 and IL-1B only in CD4+ T cells co-cultured with cancer cells. (D) The highly elevated intracellular levels of IL-6 and IL-1B were found only in T cells co-cultured with cancer cells (T15M). (A) The data from 3 independent biological replicates (donors) are presented. (B,C) The data are from 1 representative biological replicate, the other biological replicates are presented in Figure S7. The statistical significance was calculated using Student’s t-test p < 0.05 and statistically significant values are indicated by asterisks. The uncropped blots and molecular weight markers are shown in Supplementary Materials.

Fig 5: Cancer-cell-induced CD4+ T cell exhaustion may be reversed by removal of cancer cells. (A) PD-1 and PD-L1 protein levels and (B) PD-L1 expression in CD4+ T cells (T12) stimulated for 12 days after restimulation for 3 days with (T15M) and without (T15) cancer cells. Further T15M CD4+ T cells were removed from cancer cells and growing for next 3 days without (T18-M) and with (T18+M) cancer cells. (C) Expression of SWI/SNF subunits, (D) interleukins IL-1B and IL-6, and (E) angiogenesis-related encoding genes in T12, T15, T15M, T18-M, and T18+M CD4+ T cells. (B,C) The data from 3 independent biological replicates (donors) are presented. (D,E) The data are from 1 representative biological replicate. The statistical significance was calculated using Student’s t-test p < 0.05 and statistically significant values are indicated by asterisks. The uncropped blots and molecular weight markers are shown in Supplementary Materials.