L-arginine is required for expression of the activated macrophage effector mechanism causing selective metabolic inhibition in target cells.

Abstract

L-Arginine is required for expression of the activated macrophage cytotoxic effector mechanism that causes inhibition of mitochondrial respiration, aconitase activity, and DNA synthesis in tumor target cells. This effector mechanism is active in the presence of l-arginine even when the cocultivation medium lacks all other amino acids and serum. Cytotoxic activated macrophage-induced inhibition of mitochondrial respiration in target cells is proportional to the concentration of l-arginine in the medium. l-Arginine must be present during the cocultivation period. Pretreatment of cytotoxic activated macrophages with l-arginine or posttreatment of the target cells after cocultivation is not effective. D-Arginine does not substitute for l-arginine and at high concentrations is a competitive inhibitor of the l-arginine-dependent effector mechanism. Other analogues that could not replace l-arginine include agmatine, argininic acid, arginine hydroxamate, and tosyl-l-arginine methyl ester. l-homoarginine, however, can effectively substitute for l-arginine. NG-monomethyl-l-arginine is a potent competitive inhibitor of this effector mechanism. High concentrations of lipopolysaccharide do not reverse inhibition of the l-arginine-dependent effector mechanism by NG-monomethyl-l-arginine. However, inhibition of the effector mechanism by NG-monomethyl-l-arginine can be overridden by increasing the concentration of l-arginine in the culture medium. We compared NGNG-dimethyl-l-arginine and NGN1G-dimethyl-l-arginine with NG-monomethyl-l-arginine as inhibitors of the l-arginine-dependent effector mechanism. The results show that the inhibitory effect of these guanidino methylated derivatives of l-arginine is highly determined by structure. Guanidine is a weak competitive inhibitor of the l-arginine-dependent effector mechanism. The requirement for l-arginine does not appear to be for protein synthesis, creatine biosynthesis, polyamine biosynthesis, or ADP ribosylation reactions. Bacterial lipopolysaccharide is effective as a second signal only when the cocultivation medium contains l-arginine, and this strict l-arginine dependency is not overridden by increasing the concentration of lipopolysaccharide. Bovine liver arginase, by competing for l-arginine in the cocultivation medium, inhibits the l-arginine-dependent activated macrophage cytotoxic effector mechanism.