Fig 2: Immunopreventive blockade of IL-1β modulates the TME to an antitumor immune phenotype.(A) Representative flow cytometry analysis of CD8+ T cells and percentage of CD8+ T cells in the whole lung (n = 5). (B) Representative flow cytometry analysis of CD3+ programmed cell death 1–positive (PD-1+) T cells and percentage of CD3+PD-1+ T cells in the whole lung (n = 3–4). (C) Representative flow cytometry analysis of regulatory Tregs and percentage of Tregs in the whole lung (n = 4–5). (D–F) Relative mRNA expression of (D) Cd8a, (E) Gzmb, and (F) Pdcd1 in the whole lung, normalized to Cd45 or Actb expression (n = 7–9). (G and H) MFI of (G) IFN-γ and (H) granzyme B measured by multiplex ELISA in IgG or anti–IL-1β Ab–treated mice (n = 7–8). (I) Representative photomicrographs and (J) quantification of IF staining with anti-CD8a (red) and DAPI (blue) in tumor lesions (left top and bottom: original magnification, ×20, scale bar: 50 μm; right top and bottom: ×40, scale bar: 20 μm.) n = 7–8. (K) Correlation between IL1B expression and infiltration of CD8+ T cells in LUAD (n = 535). (L) Correlation between IL1B expression and PDCD1 expression in LUAD (n = 535). (M) Correlation between IL1B expression and PDCD1 expression in KM-LUAD (n = 139). Data represent mean ± SEM. *P < 0.5 by unpaired t test. a IL-1β, anti–IL-1β; TPM, transcript count per million.

Fig 3: Intestinal immune cell composition. Representative immunostaining images showing intestinal T lymphocytes (A,B) and macrophages (C,D) in ileum tissue sections from the different study groups. Black line indicates 20 µm magnification. Abbreviations: Math1Δintestine, Paneth cell depleted; PPVL, partial portal vein ligation; CD3, cluster of differentiation 3; ns, not significant. Data are expressed as mean ± SD. n = 3–5 per group. * p < 0.05; ** p < 0.005; **** p < 0.0001.

Fig 4: Loss of fibroblast STAT3 results in an increase in T-cell infiltration in KPF tumors.(A) Dual color IF of CD3 (orange) and CD8 (teal) on PDAC tumors from STAT3WT (n = 5) and STAT3cKO (n = 6) mice. (B) Dual color IF of CD4 (orange) and Foxp3 (teal) on PDAC tumors from STAT3WT (n = 5) and STAT3cKO (n = 6) mice. (A, B) Phenotype analysis generated by inFORM software is depicted in the fourth panel; yellow dots indicate double-positive cells for CD3/CD8 (A) or CD4/Foxp3 (B). (C, D, E, F) Quantification of IF using inFORM software for phenotyping and quantification of multispectral images. Error bars represent mean ± SD. Bars 50 μM. (*)P < 0.05 (***)P < 0.001 (****)P < 0.0001.

Fig 5: Intercellular communication networks among cell types in the ileum of NSPs and PWPs. a Network graph showing interactions in NSP (right panel) and PWP (left panel) between two cell types coded in colored circles. Networks depicting clusters as nodes and interactions as edges. The sizes of each cluster type are proportional to the total interaction count in each cell type, and edge thickness is proportional to interaction number between connecting types. See Additional file 15: Table S8 for detailed information. b Heatmap showing the number of ligand–receptor pairs across cell subgroups from NSPs (right panel) and PWPs (left panel), predicted by CellphoneDB. Cell types are grouped by broad lineages (intestinal epithelial cells, T cells, B cells, or other immune cells). c The recruitment of CD3+ T cells, identified by immunofluorescence (IF) in the ileum villus. Top: representative IF image showing CD3+ (green) expression in NSPs and PWPs. The numbers of CD3+ T cells significantly increased in PWPs compared with NSPs (n=6). Scale bar represents 100 μm. Bottom: the number of CD3+ cells in the field of each villus (***p<0.001, Student’s t-test; error bars: SEM). d Bubble plots showing significant ligand/receptor pairs between dendritic cells (DCs) and another cell types in NSPs and PWPs. Some selected DC–enterocyte (EC) and DC–cytotoxic T lymphocyte (CTL) interactions were enriched in PWPs but absent in NSPs. Point sizes indicate the permutation p value (CellPhoneDB). Colors reflect expression levels of ligands and receptors