The macrophage-activating tetrapeptide tuftsin was able to activate, in a dose-dependent manner, murine macrophages to express nitric oxide (NO) synthase and to produce NO. Tuftsin required lipopolysaccharides for the optimal induction of NO production and synergized with gamma interferon in the induction of NO synthesis. Tuftsin-dependent NO production was sensitive to inhibition by dexamethasone and the NO synthase specific inhibitor LGN-monomethylarginine (L-NMMA). Murine peritoneal macrophages activated by tuftsin were able to kill the amastigotes of the intracellular protozoan parasite Leishmania major in vitro.