The TME of B16 melanoma-bearing mice that were given voluntary wheel running for four weeks before tumor inoculation showed a significant change from a pro-tumoral to an anti-tumoral profile, along with the upregulation of immune checkpoint molecules such as PD-1, PD-L1, PD-L2, CD28, B7.1, and B7.2. This evidence concerns the gene CD274 and neoplasm.