The programmed death (PD)-1/PD-ligand (PD-L) pathway and regulatory T cells (Tregs) are essential for the maintenance of immune tolerance. Their activation in the tumor microenvironment contributes to the evasion of the transformed cells from the immune surveillance and the suppression of an antitumor immune response. Therefore, PD-1/PD-L1 and Tregs are important targets for cancer immunotherapy. Our review focuses on the current role of the PD-1/PD-L1 axis in Treg development and function in the tumor microenvironment. We also discuss combination therapy with PD-1/PD-L1 inhibitors and Treg-modulating agents affecting the adenosinergic pathway, TGF-β signaling, immune checkpoints, and other approaches to downregulation of Tregs.