Changes in chemokine expression and immune cell abundance in CD206-depleted tumors, and associated survival data in humans . (A) Dotplot representing top five DEGs and select other genes in each immune cell cluster identified from a harmonized dataset of spatially barcoded Control and DTx treated B78chOVA tumors on day 12 after adoptive transfer of CD2dsRed; OT-I cells. (B and C)Cxcl10 expression (B) aggregated across treatment conditions by cluster and (C) aggregated across clusters by treatment. (D) CXCL9 expression in PyMTchOVA and MC38chOVA (both without OT-I adoptive transfer) TAMs split by their CD206 expression. (E and F) (E) CXCL9 expression in B78chOVA intratumoral monocytes split by CD206 expression and (F) relative abundance of CXCL9⁺ TAMs and monocytes in the same tumors. (G) Flow cytometry plot showing CD206 and CXCL9 expression in cDC2s and TAMs in B78chOVA tumors. (H) Percent of CXCR3⁺ among in vitro activated CD8 T cells prior to transmigration in the presence of CD206⁻ and CD206⁺ TAMs. (I and J) Violin plot representing (I) Cxcr3 and (J) Xcl1 and Flt3l expression in the lymphoid compartment in Control and DTx treated conditions. (K) Tumor growth curves of B78chOVA tumors with OT-I adoption and early and late DTx administration. (L and M) (L) Overlaid flow cytometry plots showing reporter (Venus) and CD206 expression in different immune cells in MC38chOVA tumors in WT (red; Mrc1^{LSL-Venus-DTR}) and DTR (blue; Csf1r^Cre; Mrc1^{LSL-Venus-DTR}) mice and (M) quantification of relative reporter expression (DTR – WT) in the different subsets, split by CD206 expression. (N) Schematic representation of the experimental setup for early and late CD206⁺ TAM depletion in MC38chOVA tumors using Mrc1^{LSL-Venus-DTR} (WT) and Csf1r^Cre; Mrc1^{LSL-Venus-DTR} (DTR) mice. (O and P) Relative abundance of different immune populations as a percentage of CD45⁺ cells with (O) late and (P) early depletion regimens. (Q) Abundance of different immune populations as total number of cells per g of MC38chOVA tumor in WT and DTR mice in the early DTx administration regimen. (R) Number of CD8 T cells and NK cells per tdLN of WT and DTR mice with MC38chOVA tumors and treated with the early DTx regimen. (S) Scatter plots of the CD206^Replete and CD206^Depleted Mono/Mac score per patient with the NK cell score (Pearson R and P value for the null hypothesis that there is not a correlation are noted). (T) Kaplan–Meier survival curves of patients grouped by the value of the CD206^Replete: CD206^Depleted signature ratio (top and bottom 20%) from TCGA split by indications, number of patients per group and P values for the log-rank test are noted for each curve in T. Bar graphs show mean ± SEM; data are representative of at least two independent experiments, each with at least three biological replicates, except the spatial transcriptomics data (A–C, I, and J), from one control and one DTx-treated tumor. ***P < 0.0001, **P < 0.01, *P < 0.05, ns = no significance by paired ratio t tests (D and E) or unpaired t tests or Mann–Whitney test in F and O–Q. n = 203 patients in S.