Glioma-derived soluble factor(s) enhance IL-10 signaling.

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Glioma-derived soluble factor(s) enhance IL-10 signaling. Glioma-derived soluble factor(s) enhance IL-10 signaling. A, human IL-10 expression measured by ELISA from culture media of normal monocytes stimulated with glioma-conditioned media before and after 24 hours of culture. None of the conditioned media initially contained appreciable levels of IL-10. After 24 hours, IL-10 expression was significantly increased in cells stimulated with glioma-conditioned media compared with normal media (*, P < 0.01). Columns represent mean total IL-10 ± SEM from 3 independent samples. B, IL-10 receptor expression was measured by flow cytometry in monocytes stimulated with glioma-conditioned media. Compared with unstimulated cells, treated monocytes had a modest but significant increase in IL-10R mean fluorescence intensity (*, P < 0.05; **, P < 0.01). Columns represent mean fluorescence intensity ± SEM from 3 independent samples. Each sample was tested in triplicate and averaged as a single data point. C, time course of IL-10 and B7-H1 mRNA expression in normal monocytes stimulated with SF1U-conditioned media. Transcript levels were measured at 0, 1, 6, 12, and 24 hours after stimulation using quantitative RT-PCR and normalized to 18S rRNA. Normalized expression is reported as a relative increase over initial expression. Each point represents mean relative expression ± SEM from 3 independent samples. D, representative section of GBM tissue with fluorescent immunestaining for CD163 (red), B7-H1 (green), IL-10 (cyan), and 4′, 6–diamidino–2–phenylindole (DAPI; blue) showing colocalization of all 3 probes to the same subset of tumor-associated macrophages (scale bar = 10 μm). Orin Bloch et al. Clin Cancer Res 2013;19:3165-3175 ©2013 by American Association for Cancer Research