Pharmacologically attainable concentrations of ascorbic acid are highly toxic for U937 cells (a human promyelocytic cell line), and this response appears to be mediated by H2O2. This inference finds experimental support in the following observations: 1) toxic levels of H2O2 are readily generated upon dissolution of survival-range concentrations of ascorbic acid in the culture medium; 2) the lethal effects elicited by ascorbic acid or reagent H2O2 are prevented by the addition of either catalase or the intracellular iron chelator o-phenanthroline and are characterized by similar temporal dependence; 3) U937 cells resistant to hydrogen peroxide are cross-resistant to ascorbic acid; 4) under the conditions utilized in this study, H2O2 and ascorbate promote similar modes of cell death (i.e., necrosis); and 5) cell killing provoked by H2O2 or ascorbate is an inverse function of cell density and is suppressed by coculturing U937 target cells with human erythrocytes (at a density far below that present in the blood) and human fibroblasts. Cytoprotection was not observed using catalase-depleted erythrocytes. Taken together, these results strongly suggest that H2O2 is entirely responsible for the ascorbate-induced U937 cell killing. We therefore propose that it is unlikely that the vitamin damages or kills tumor cells of normal tissues in vivo via the H2O2 based mechanism, because the oxidant would be removed promptly by the neighboring cells.