Fig. 5
DMXAA reprograms TAMs in vivo and activates macrophages in vitro. Tumors were harvested on day 19, dissociated and immune cell subsets analyzed by flow cytometry. a Percent of viable CD45+ leukocytes, CD11b+,Ly6G−, Ly6CLo, F4/80Hi, MHC Class II+ myeloid tumor-associated macrophages (TAM), and CD206hi TAMs. b Mean fluorescence intensity (MFI) of CD86 and PD-L1 levels expressed on CD45+CD11b+ TAMs from control (NT, open bar) or DMXAA [450 μg] (black bar) treated mice. Values are mean ± SD, n = 6 mice per group. *, P ≤ 0.05; **, P ≤ 0.01. c Cultured bone marrow-derived macrophages were untreated or treated for 18 h with 20 μg/ml DMXAA. Histograms from representative flow cytometry analyses for CD80, CD86, CD40, MHC class I, PD-L1 and CD206 expression on bone marrow-derived macrophages from non-treated (black line) DMXAA-treated (black area) and compared against unstained cells (gray dotted lines). d, e Multiplex analyses of cytokine and chemokine production measured in conditioned medium from bone marrow-derived macrophages cultured 18 h in the presence (black bars) or absence (open bars) of 20 μg/mL DMXAA. Values are mean ± SD of two combined experiments. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001