Fig 2: 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 3: Immunotherapeutic blockade of IL-1β increases activated T cells’ infiltration.(A) Total inflammatory cell and lineage-specific leukocyte numbers from BALFs of 18-week-old CC-LR mice treated with IgG or anti–IL-1β Ab (n = 4–5). (B) Representative flow cytometry analysis of neutrophils (CD11b+Ly6G+) and PMN-MDSCs (CD11b+Ly6CloLy6G+) in BALFs. (C and D) Quantification percentage of (C) neutrophils and (D) PMN-MDSCs in BALFs (n = 5–6). (E–L) Representative flow cytometry analysis and quantification percentage of CD3+PD-1+ T cells (E and F), CD8+ T cells (G and H), IFN-γ–expressing CD8+ T cells (I and J), and granzyme B–expressing CD8+ T cells (K and L) in whole lungs of 18-week-old CC-LR mice treated with IgG or anti–IL-1β Ab (n = 3–6). (M–O) Relative mRNA expression of Ym1 and Mrc1 (M), Cd8a (N), and Pdcd1 (O) in the whole lung, normalized to Cd45 or Actb expression (n = 3–5). (P and Q) MFI of TNF-α (P) or granzyme B (Q) measured by multiplex ELISA in IgG or anti–IL-1β–treated mice (n = 4–5). Data represent mean ± SEM. **P < 0.01, *P < 0.5 by unpaired t test. a IL-1β, anti–IL-1β; Lym, lymphocytes; Mac, macrophages; Neu, neutrophils.

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