In this in vitro study, an evaluation of the importance of intracellular oxidative balance on cell-mediated cytotoxicity was performed by analyzing the effects of the antioxidant N-acetylcysteine (NAC), a specific thiol supplier, on natural killer (NK) cell-mediated cytotoxicity. The results obtained indicate that an enhancement of target cell (TC) killing can be detected when a pre-exposure of effector cells (EC) to NAC was performed. However, this effect seems to depend upon the TC type used. In fact, the increase of EC activity was detected against the differentiated U937 TC while no changes were detected by the same effectors against K562 cells. The mechanism of this enhancement seems to be ascribable to an increased ability of NAC-exposed NK cells to form conjugates (binding) which, in turn, appears to be due to a specific effect of NAC on actin microfilaments. A role for NAC as a cytoskeleton thiol-modifier contributing to the activation of effector cells can thus be hypothesized.