Fig. 6: Kupffer cells and T cells coordinate BMDM activation.

a Study design for (b–j). b UMAP of liver populations identified by scRNAseq in control (n = 3) and BG (n = 3)-treated livers. c Dot plot of Log₂ Fold-Change in liver population frequencies in BG-treated livers compared to control livers. Size represents proportion of total cells and color indicates number of differentially expressed genes in BG versus control. d UMAP of BMDM/cDC populations identified by scRNAseq. e Dotplot of Log₂ fold change in BMDM population frequencies in BG-treated livers compared to control livers. Size represents proportion of total cells and color indicates number of differentially expressed genes in BG versus control. f Radar plot showing selected gene sets expressed by Spp1⁺MΦs in control (blue) and BG (orange) treated livers. Axis represents the mean TPM of selected gene set-associated genes with range 0 to 172 TPM. g, h Heatmaps of immune regulatory (g) and nitric oxide-related (h) gene set associated genes, upregulated in Spp1⁺MΦs from control and BG-treated livers. Scale bars indicate log₂[TPM] scaled by row across samples. i, j Violin plots of Cd274 (i) and Nos2 (j) expression by Spp1⁺MΦs in control and BG-treated livers. k Representative images of livers from control mice or mice treated with BG, BG + DT, BG + CEL, or BG + a4/8 stained using IHC to detect NOS2 (brown) and nuclei (blue, hematoxylin). The bottom panel highlights the lesion interior. Scale bars, 100μm (top) and 50 μm (bottom). Metastatic lesions, blue dashed lines. l Quantification of NOS2⁺ staining detected by IHC and shown as percentage of individual control (n = 248), BG-treated (n = 173), BG + DT (n = 350), BG + CEL (n = 260) and BG + α4/8 (n = 61) liver lesion area. Data represents the combination of three independent experiments with control (n = 5), BG-treated (n = 6), BG + DT (n = 8), BG + CEL (n = 5), and BG + α4/8 (n = 5) biological replicates. One-way ANOVA with Sidak’s multiple comparisons test. Mean ± SD is shown. iPo, intraportal; KC, Kupffer cell; BG, β-glucan; scRNAseq, single cell RNA sequencing; MΦ, macrophage; cDC, conventional dendritic cell; Ctrl, control; DT, diphtheria toxin; CEL, clodronate encapsulated liposomes; a4/8, anti-mouse anti-CD4/anti-CD4 monoclonal antibody.