Substance P-induced IL-12 production by murine macrophages

Previous investigations in our laboratory have suggested that substance P (NK-1) receptor expression by macrophages contributes to the resistance against the intracellular bacterial pathogen, Salmonella. To investigate possible mechanisms for such resistance, macrophages were cultured with varying concentrations of a substance P agonist to investigate the ability of this neuropeptide to augment IL-12 expression. The substance P agonist was a potent inducer of both IL-12p35 and IL-12p40 mRNA expression in cultured macrophages. The kinetics of this response were maximal within 6 h and could be observed with concentrations of substance P agonist as low as 0.1 nM. The nonpeptide, substance P receptor antagonist, CP96-345, significantly blocked agonist-induced IL-12 mRNA expression, further demonstrating that this effect was mediated through an NK-1 receptor. Substance P agonist alone could stimulate substantial secretion of IL-12p40, but not IL-12p70, by cultured macrophages. Thus, the substance P agonist had the ability to augment IL-12p35 and IL-12p40 mRNA expression, but not to increase IL-12p70 secretion. Like IFN-gamma, we found that substance P could combine with LPS to significantly augment the secretion of bioactive IL-12p70. The costimulatory effects of substance P agonist plus LPS on IL-12 mRNA expression were additive; however, this combination resulted in synergistic secretion of IL-12p70 by macrophages. Together, these results demonstrate the ability of NK-1 receptors to signal IL-12 production by macrophages and suggest mechanisms for substance P-induced modulation of cellular immunity.