Several non-phagocytic cells can actively generate the superoxide anion by NAD(P)H oxidases resembling the enzymatic complex typical of phagocytes. Overexpression of periplasmic Cu,ZnSOD rescues invasive E. coli strains from killing within epithelial cells, suggesting that superoxide generation by such cells can oxidatively damage invading bacteria. Pre-treatment of HeLa cells with diphenyl iodonium or 4'-hydroxy-3'-methoxyacetophenone, two inhibitors of NAD(P)H oxidase, significantly enhances intracellular survival of wild type invasive E. coli cells. On the contrary, these inhibitors have no effect on the intracellular survival of an invasive E. coli strain engineered to overexpress Cu,ZnSOD. These results support the hypothesis that superoxide generation by a NAD(P)H oxidase-like complex can limit bacterial survival within epithelial cells and suggest that the role of periplasmic Cu,ZnSOD in bacterial infections is not simply that of conferring protection against the phagocytic oxidative burst.