Fig 2: BRCA1-deficient breast tumor cells induce M2-like macrophage polarization in vitro.a Diagram of workflow. Top, mouse bone marrow-derived macrophages (BMDMs) co-cultured with tumor cells with or without olaparib treatment. Bottom, BMDMs were incubated with 50% conditioned media (CM) harvested from olaparib- or DMSO-treated tumor cells. TEMs, tumor cell-educated macrophages. b Flow cytometry analysis of BMDMs co-cultured with BP tumor cells with or without olaparib (5 µM) for 2 days. BMDMs (CD11b+) were plotted as CD206 versus MHC-II to identify M1-like (CD206- MHC-IIhigh) and M2-like (CD206+ MHC-IIlow) polarization phenotypes (BMDMs, n = 3; BP/BMDMs, n = 7). Flow cytometry plot axes are displayed in logarithmic scale. c Heat map of gene expression for anti-tumor and pro-tumor genes in BMDMs incubated with DMSO vehicle control, olaparib (OL, 5 µM), 50% BP-CM, or 50% OL-treated BP (BP/OL)-CM for 24 h (n = 2 for each group). d RT-qPCR analysis of mouse BMDMs incubated with control medium, 50% BP-CM, or 50% BP/OL-CM for 24 h (Il6, n = 5; Il1b, n = 3; Cxcl1, n = 4). e RT-qPCR analysis of THP-1 human macrophages incubated with control medium, 50% tumor cell-CM, or 50% CM of olaparib-treated tumor cells for 24 h (MDA-MB-436, n = 5; HCC1937, n = 4). Data are presented as mean ± SEM. One-way ANOVA (b). Two-tailed unpaired t test or Mann–Whitney test (d) and (e). ns, not significant. Source data are provided as a Source Data file.

Fig 3: CD11c+ cells express CXCL16 in melanoma-associated vitiligo (MAV)–affected skin.(A) Heat map showing gene expression of chemokines and receptors in CD11c+CD11bneg, CD11c+CD11b+, and CD8+ T cells sorted from skin of MAV-affected mice. Data presented as log2-normalized expression. (B) Surface expression of CXCL16 on CD11c+ cells isolated from skin of naïve (n = 3) or MAV-affected (n = 5) mice, skin-draining LNs, or spleen 7, 14, and 35 d after surgery. Representative of three independent experiments. (C) Surface expression of CXCL16 on CD45+CD11c+, CD45+CD11cneg, and CD45neg cells isolated from skin of MAV-affected mice, 35 d after surgery. Representative of two independent experiments with skin collected between 35- and 60-d post-surgery. Dotted horizontal line indicates background CXCL16 expression in unstained cell sample. (D) Expression of CXCL16 and CD11c in unaffected and MAV-affected mice; CD11c (green), CXCL16 (red), and nuclei (blue). Scale bar, 50 µm. (E) CXCL16 expression in IF images from skin of unaffected and MAV-affected mice; n = 5–8 mice. (F) CXCL16 expression in hair follicles with or without CD11c+ cell clusters in MAV-affected skin; n = 8 mice. (G) CD11c+ and CXCL16+ cells in skin from MAV-affected patients. Stains identify CXCL16 (green) and CD11c (red); dark red identifies colocalization of CD11c and CXCL16. Scale bar, 25 µm. (H) Percent of CD11c+ cells expressing CXCL16 in MAV-affected patient skin. (B, C, H) Symbols represent individual mice (B, C) or patients (H); horizontal lines indicate mean. MFI, mean fluorescence intensity. Significance was determined by one-way ANOVA; *P = 0.03, ***

Fig 4: Brca1-deficient breast tumors have a modest response to olaparib in vivo with immune-suppressive TAMs.a Generation of a syngeneic GEMM of Brca1-deficient breast tumors by intraductal injection of adenovirus expressing Cre recombinase (Ad-Cre) directly into the lumen of mammary glands. b Tumor-free survival of Brca1L/L Trp53L/L mice with or without intraductal injection of Ad-Cre (n = 6). c Tumor growth of Brca1-/- Trp53-/- (BP) allografts in FVB mice treated with olaparib or anti-PD-1 as monotherapy or in combination. Control, n = 8; anti-PD-1, n = 8; olaparib, n = 6; olaparib + anti-PD-1, n = 10. d Flow cytometry analysis of TCRß+ T cells and tumor-associated macrophages (TAMs; CD45+ CD11b+ F4/80+) from BP tumors in FVB mice after 21 days of olaparib treatment. TCRß+ (control), n = 6; TCRß+ (olaparib), n = 8; TAMs (control), n = 6; TAMs (olaparib), n = 6. TAMs were further analyzed to identify M1-like (MHC-IIhigh CD206-) and M2-like (MHC-IIlow CD206+) polarization phenotypes (n = 6 for each group). Each dot represents data from a single tumor. e Diagram of workflow for (f) and (g). TAMs (7AAD- CD45+ CD11b+ F4/80+) were sorted from BP breast tumors and co-cultured with splenic CD8+ T cells isolated from naïve mice for 2 days. f Analysis of cytokine production by CD8+ T cells co-cultured with TAMs (CD8+ T cells, n = 4; CD8+ T cells+TAMs, n = 8). g Analysis of the proportion of effector cells (CD44high CD62Llow) and surface expression of CD25 of CD8+ T cells co-cultured with TAMs (CD8+ T cells, n = 4; CD8+ T cells+TAMs, n = 8). Flow cytometry plot axes are displayed in logarithmic scale (f) and (g). Data are presented as mean ± SEM (c), (f), and (g), or median with quartiles (violin plots, d). Two-way analysis of variance (ANOVA) (c). Two-tailed unpaired t test or Mann–Whitney test (d), (f), and (g). ns not significant. Source data are provided as a Source Data file.

Fig 5: Combination LEF+MTX suppresses myeloid cell adhesion to the sinusoidal endothelium and hinders fibrosis resolution. (A) Representative images of the liver sinusoidal endothelium illustrating the CD11b+ cells (green). (B) CD11b+ cells per liver sinusoid were calculated. (C) Representative images of Masson staining. (D) mRNA and (E) protein expression levels of VEGF-A were detected by reverse transcription-quantitative polymerase chain reaction and ELISA, respectively. Data are presented as the mean ± standard deviation (n=6). *P<0.05, **P<0.01 and ***P<0.001. Scale bar, 200 µm. CCl4, carbon tetrachloride; CD, cluster of differentiation; LEF, leflunomide; MTX, methotrexate; NC, normal control; VEGF, vascular endothelial growth factor.