Induction of iron-derived EPR signals in murine cancers by nitric oxide. Evidence for multiple intracellular targets.

The cell-mediated immune response to syngeneic tumors activates the cytokine-inducible nitric oxide synthase. We observed that syngeneic murine tumors exhibited EPR signals related to iron-nitrosyl complex formation. Three different EPR active iron-nitrosyl species were observed, an Fe(RS)2(NO)2 signal and two differentiable heme-nitrosyl complexes. Hemoglobin assays showed that the heme-nitrosyl signals were not derived from contaminating hemoglobin. Signal amplitudes were attenuated in mice treated with N omega-mono-methyl-L-arginine (MLA), an inhibitor of nitric oxide synthase. Tumors grown in vivo contained EPR signals while those grown in culture without continuing cytokine stimulation lost the signals after a few days. Cultured cells that were treated with cytokines, or that were cocultivated with cytokine-activated macrophages, regained EPR active complexes. These results show that the cell-mediated immune response to syngeneic tumors involves the induction of nitric oxide synthase. While nitric oxide synthesis is induced in both tumor infiltrating macrophages and in the tumor cells themselves, only tumor cells contributed to formation of heme-nitrosyl complexes. This result indicates the presence of a novel intracellular target for NO within tumor cells.