We have reported that paclitaxel (PCT), an anti-cancer agent, causes perineuronal accumulation of macrophages (Mφ), which in turn secret high mobility group box 1 (HMGB1), a nuclear protein, participating in the development of chemotherapy-induced peripheral neuropathy (CIPN) in mice. Given our previous evidence that neuron-derived “unknown molecules” facilitate HMGB1 release from Mφ, we examined whether ATP would participate in the neuroimmune crosstalk involved in CIPN caused by PCT. In Mφ-like RAW264.7 cells, PCT at 100-1000 nM, but not ATP at 100 μM, caused extracellular release of HMGB1. Combined application of ATP at 100 µM and PCT at 50 nM, a subeffective concentration, induced HMGM1 release from RAW264.7 cells. PCT at 50 nM caused ATP release from neuron-like NG108-15 cells. In a Transwell co-culture system of RAW264.7 and NG108-15 cells (upper and lower chambers, respectively), PCT at 50 nM increased the amount of HMGB1 in the medium of both chambers, an effect prevented by A438079 and 5-BDBD, P2X₇ and P2X₄ receptor antagonists, respectively. Finally, we found that CIPN in mice treated with PCT was suppressed by A438079 or 5-BDBD. Our data thus suggest that neuron-derived ATP promotes PCT-induced HMGB1 release from Mφ, contributing to the development of CIPN in mice treated with PCT.