In vivo cisplatin-exposed macrophages increase immunostimulant-induced nitric oxide synthesis for tumor cell killing.

Mice pre-exposed to cisplatin increased their production of nitric oxide (NO) when treated with lipopolysaccharide (LPS). Peritoneal macrophages, isolated from mice 11 days after cisplatin treatment and cultured with LPS plus IFN-gamma, increased NO production, whereas the macrophages isolated 2 days after cisplatin treatment decreased it. In both cases, NO was not produced without immunostimulant(s). Northern and Western Blot analysis showed that macrophages exposed to cisplatin for 11 days increased production of mRNA and protein expression of inducible nitric oxide synthase (iNOS). THis result indicated that macrophages became more sensitive to LPS and IFN-gamma when they were exposed to cisplatin in vivo. Peritoneal macrophages, when activated with LPS plus IFN-gamma and then cocultured with several tumor cells, exhibited cytotoxic activity against both cisplatin-sensitive and cisplatin-resistant tumor cells. There was no difference in cytotoxicity between the paired cells. Under the same experimental condition, macrophages that were exposed to cisplatin for 11 days had significantly increased their cytotoxicity to the tumor cells. This cytotoxic activity was inhibited by the NOS inhibitor NG-monomethyl-L-arginine, indicating that NO is a major effector for macrophage-mediated tumor cell killing. Treatment of tumor cells with S-nitroso-N-acetylpenicillamine, a NO-generating compound, showed the similar tumoricidal effect. These data demonstrated that injection of cisplatin into the mice can enhance the sensitivity of macrophages to the subsequent treatment of immunostimulant(s) for effective tumor cell killing through enhanced NO production.