Panel A shows a uniform manifold approximation and projection (UMAP) plot and dot plot of CD1D expression across human breast cancer cell types. Panel B shows histogram plots comparing CD1d expression in WT, CD1d-knockout, and EO771 cells. Panel C shows histogram plots and a bar plot of CD1d geometric mean fluorescence intensity across myeloid cell populations. Panel D shows bar plots comparing immune cell populations per cubic centimeter of tumor in WT mice. Panel E shows a line graph of tumor size in cubic millimeters and a bar plot of tumor weight in grams. Panel F shows bar plots of live cells and cell numbers after anti-CD1d treatment at different antibody concentrations. Panel G shows bar plots comparing myeloid and dendritic cell populations in CD1d-knockout mice after treatment. Panel H shows a line graph of tumor size in cubic millimeters and bar plots of immune cell populations and granzyme B positive CD8 positive T cells.
Anti-CD1d regulates immune infiltrates and tumor growth. (A) Left: UMAP showing CD1D expression in cell clusters from human breast cancers (Wu et al., 2021). Right: Dot plot showing the expression of CD1D in the depicted populations of human breast cancers. (B) Flow cytometry plot depicting CD1d expression in WT and CD1d-KO BMDMs and in EO771 cells as indicated. Data are representative of two independent experiments. (C) Flow cytometry plot for CD1d expression in the depicted myeloid cell populations identified in the EO771 TME in WT mice. Right: GeoMFI for the depicted populations. Data are representative of two independent experiments. (D) Frequencies of the indicated cell populations in EO771 tumors in WT mice receiving αCD1d (or isotype) as indicated in Fig. 2 C. (E) Tumor growth in WT mice orthotopically injected with EO771 cells and receiving αCD1d (or isotype) at the indicated time points (arrows). Data are shown as the mean ± SEM. The bar plot represents tumor weight at day 15 (n = 8; data are pooled from three independent experiments). (F) EO771 cells were cultured in vitro with increasing concentrations of αCD1d (or isotype) for the indicated time points. Graphs show frequency of alive cells after 24 h (left) and number of cells (right) at different time points after αCD1d culture. Data are representative of two independent experiments. (G) Frequencies of the depicted cell populations in EO771 tumors from CD1d-KO mice receiving αCD1d (or isotype) at the time points indicated in Fig. 2 D. Data are shown as the mean ± SEM. (H) Tumor growth in WT BALB/c mice orthotopically injected with 4T1 cells and receiving αCD1d (or isotype) at the indicated time points (arrows). Bar plots represent frequencies of the indicated cell populations at day 10 (n = 5; data are pooled from two independent experiments). Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, unpaired (D, E, and H) t test or two-way ANOVA with Sidak’s (E, H, tumor growth plot) multiple comparisons. GeoMFI, geometric mean fluorescence intensity.