Peptidoglycans (PGN) from bacterial cell walls may modify the course of an infection with bacterial pathogens. The present study explored the effect of PGN on cytosolic Ca²⁺ activity, cytokine production and phagocytosis of mouse dendritic cells (DCs), essential cells in the initiation and direction of antigen-specific T cell responses. Exposure of DCs to PGN was followed by a rapid increase in cytosolic Ca²⁺ activity ([Ca²⁺]_i), which was due to Ca²⁺ release from intracellular stores and influx of extracellular Ca²⁺ across the cell membrane. The PGN-induced increase of [Ca²⁺]_i was dependent on voltage-gated K⁺ (Kv) channel activity. PGN-induced increase of [Ca²⁺]_i was significantly blunted by margatoxin (MgTx) and perhexiline maleate (PM), both inhibitors of Kv1.3 and Kv1.5, respectively. PGN further stimulated the release of TNFa and IL-10, both effects significantly blunted by MgTx, PM and the specific blocker of store-operated Ca²⁺ channels SKF-96365. Moreover, phagocytic capacity was dramatically increased in PGN-stimulated DCs in the presence of either Kv channel inhibitors or SKF-96365. The observations disclose Ca²⁺- and Kv-channel-dependent cytokine production and phagocytosis in PGN-stimulated DCs.