Effect of inhibition of CCL3-CCR1 axis on macrophage programming. (A-B) THP1- and U937-MΦ were cocultured with Mino cells and treated with or without 1μM CCR1 inhibitor or 0.5 μg/mL of CCL3 neutralizing antibody for 48 hours and expression of CD206 and IL-10 (A) or CD86 (B) mRNA was assessed. (C) Cells from the patients with MCL were cocultured with CD14⁺ Mo and treated with or without 1μM CCR1 inhibitor or 0.5 μg/mL of CCL3 neutralizing antibody and mRNA expression of CD206, IL-10, and CD86 was evaluated using qRT-PCR. (D) THP1-MΦ were incubated with Mino or JVM2 CM with or without 1μM CCR1 inhibitor BX-471 treatment, and the secretion of IL-10 was assessed using ELISA. (E) Immunofluorescent staining was performed on the syngeneic mouse tumor–bearing FC-muMCL1 cells using IL-10 (red) and CD206 (green) antibodies. Nuclei were stained with DAPI (blue). (F-G) CD206 or CD86 and MHC II expression were assessed in WT (n = 3) or IL-10-/- BMDM (n = 3) with or without rIL-10 (100 ng) treatment using qRT-PCR. Data presented are representative of 3 independent experiments unless stated otherwise (∗P < .05; ∗∗P < .01).