Panel A shows paired scatter plots of gene expression in mouse bone marrow-derived macrophages treated with anti-CD1d or isotype and stimulated with interferon gamma or interleukin 4. The y-axis represents gene expression in arbitrary units, and the x-axis shows different treatments. Panel B shows similar paired scatter plots for human monocyte-derived macrophages. Panel C shows a schematic diagram, a line graph of tumor growth over time, and bar graphs of tumor weight and immune cell populations. The line graph y-axis represents tumor size in square millimeters, and the x-axis represents days. The bar graphs show weight in milligrams and cell counts per cubic centimeter. Panel D shows similar tumor growth and immune cell analyses for WT and CD1d-knockout mice. Panel E shows a bar graph of tumor size and a line graph of tumor growth in different mouse genotypes treated with anti-CD1d or isotype. Panel F shows a line graph of tumor growth in WT mice treated with anti-CD1d and interferon gamma. Panel G shows a schematic diagram, a bar graph of tumor weight, and a line graph of tumor growth in WT mice treated with anti-CD1d and anti-programmed cell death protein 1 (anti-PD-1). Significant differences are indicated by asterisks.
Anti-CD1d reduces tumor growth and enhances anti-PD-1–induced tumor regression. (A and B) Mouse BMDMs (A) or human MDMs (B) were cultured with αCD1d or isotype and stimulated with IFN-γ or IL-4 as indicated (or left unstimulated, u). Expression of the depicted genes was measured by qPCR (n = 5–7; data are pooled from three independent experiments). (C) WT mice were orthotopically injected with EO771 cells and received αCD1d (or isotype) at the indicated time points (arrows). Tumor growth was monitored over time (left). Bar plots represent tumor weight (middle) and numbers for the indicated cell populations (right) at day 15 (n = 5; data are pooled from two independent experiments). Data are shown as the mean ± SEM. (D) WT or CD1d-KO mice were orthotopically injected with EO771 cells and received αCD1d (or isotype) at the indicated time points (arrows). Tumor growth was monitored over time (left). Bar plots represent tumor weight (middle) and numbers for the indicated cell populations (right) at day 15 (n = 3; data are pooled from two independent experiments). Data are shown as the mean ± SEM. (E) Tumor size (left, day 10) and growth (right) in WT, Rag2-KO, or Rag2-γc-dKO mice orthotopically injected with EO771 cells and receiving αCD1d (or isotype) at the indicated time points (arrows) (n = 3–7; data are pooled from two to three independent experiments). Data are shown as the mean ± SEM. (F) Tumor growth in WT mice orthotopically injected with EO771 cells and receiving αCD1d ± αIFN-γ at the indicated time points (arrows) (n = 4–7; data are pooled from three independent experiments). Data are shown as the mean ± SEM. (G) WT mice were orthotopically injected with EO771 cells and received αCD1d ± αPD-1 (or isotype) at the indicated time points (arrows). The bar plot represents tumor weight at day 12 (n = 6; 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 (C, D, and G) or paired (A and B) t test, or two-way ANOVA with Tukey’s (E, left) or Sidak’s (C–G, tumor growth plots) multiple comparisons.