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. Author manuscript; available in PMC: 2024 Sep 21.
Published in final edited form as: Nat Med. 2024 Jun 28;30(8):2170–2180. doi: 10.1038/s41591-024-03075-7

Figure 3 |. Pre-metastatic livers of PaC patients feature changes in infiltrating immune cells.

Figure 3 |

a, Liver biopsies were stained by immunofluorescence (IF) for the myeloid marker CD11B, quantified with ImageJ, and compared between PaC (n=44) and Non-PaC (n=14; Mann-Whitney U-test; p=0.005). b-c, Liver biopsies were stained by immunohistochemistry (IHC) for the macrophage activation marker IBA-1 and compared between PaC (n=47) and Non-PaC (n=9) after manual quantification by a blinded pathologist of b, the extent of IBA-1+ cell infiltration in the portal tracts (red outline; Somers’ d; p=0.001), and c, the presence of IBA-1+ cell aggregates (yellow outline) in the lobular portions of the liver parenchyma (Somers’ d; p=0.005). d, Liver biopsies were stained for neutrophil elastase (NE) and citrullinated H3 (Ct-H3) to identify clusters consistent with neutrophil extracellular traps (NETs). Stained area was quantified with ImageJ and compared between PaC (n=22) and non-PaC (n=9; Mann-Whitney U-test: NE area, p=0.016); CtH3 area, p=0.006). e-f, Liver biopsies were co-stained by IF for CD3 and CD8. e, CD3+ T cells were quantified using ImageJ and compared between PaC (n=42) and Non-PaC (n=13; t-test; p=0.008). f, A blinded pathologist quantified the intensity of CD3+ lymphocyte staining, (Somers’ d; p=0.028). g-i, Single-cell RNAseq was performed on hepatic NPCs (>90% CD45+) from 3 non-PaC and 5 PaC patients and analyzed as shown in Ext. Data Fig. 4–5. g, Comparison of the relative abundance of the major cell clusters between PaC and Non-PaC (multiple t-tests with correction for multiple comparisons, shown if p<0.25). g, The percentage of different cell clusters among CD11B+ cells was calculated and compared between PaC and non-PaC (multiple t-tests with correction for multiple comparisons, shown if p<0.25). h, Subset analysis of CD3+ cells (Ext. Data Figure 4) resulted in four T cell subclusters as well as NKT cells and two NK cell cluster (MAIT, mucosa-associated invariant T cells). i, Subset analysis of CD3+ cells resulted in four T cell subclusters as well as NKT cells and two NK cell clusters (MAIT, mucosa-associated invariant T cells). Mean±SEM are shown in bar graphs.