The adenosine pathway was inhibited using the following agents. The A2aR inhibitor vipadenant (provided by Juno Therapeutics, a Celgene company) was suspended in 40% Captisol (Captisol Ligand Technology) and 10% (w/s) PEG400 (MilliporeSigma) in PBS, and SCH58261 (MilliporeSigma) was dissolved in DMSO (MilliporeSigma) and then diluted in Cremophor EL (MilliporeSigma) and 0.9% NaCl (final concentration, 15% DMSO and 15% Cremophor EL). These agents have been reported to have blood-brain barrier penetration in human subjects with Alzheimer’s and Parkinson’s disease (16 (link), 22 (link)–27 (link)). The mice were treated daily for 21 days, starting on day 3 after glioma implantation, via i.p. injection with either SCH58261 (10 mg/kg) or with vipadenant (60 mg/kg). The CD39 inhibitor POM-1 (Tocris) was dosed on the same schedule at 5 mg/kg. Anti-CD73 (clone TY/23; Bio X Cell; RRID: AB_10950310) or IgG control (clone 2A3, Bio X Cell; RRID: AB_1107769) was administered intravenously at a dose of 200 μg/mouse on days 3, 6, 10, 14, 17, and 21. To evaluate complementary immune suppression blockade, an anti PD-1 (clone RMP1-14; Bio X Cell; RRID: AB_10949053) or IgG control (clone 2A3, Bio X Cell; RRID: AB_1107769) was administered i.p. at a dose of 200 μg/mouse on days 7, 9, and 10.
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