THE MICROBICIDAL ACTIVITY OF INTERFERON-GAMMA-TREATED MACROPHAGES AGAINST TRYPANOSOMA-CRUZI INVOLVES AN L-ARGININE-DEPENDENT, NITROGEN OXIDE-MEDIATED MECHANISM INHIBITABLE BY INTERLEUKIN-10 AND TRANSFORMING GROWTH-FACTOR-BETA

The present study was carried out to determine the effector mechanism of anti-Trypanosoma cruzi activity by interferon (IFN)-gamma plus lipopolysaccharide (LPS)-treated macrophages. A macrophage cell line (IC-21) that failed to mount an appreciable oxidative burst was nevertheless found able to control T. cruzi growth after exposure to IFN-gamma alone or IFN-gamma plus LPS. Moreover, microbicidal functions of both inflammatory macrophages and IC-21 against T. cruzi was found to be inhibited in the presence of N(G)-monomethyl-L-arginine (N(G)MMA), a competitive inhibitor of L-arginine. Addition of supplemental L-arginine to the culture overcame the capacity of N(G)MMA to block activated macrophage anti-T. cruzi functions. The ability of N(G)MMA to reverse both parasite growth inhibition and killing by IFN-gamma plus LPS-activated macrophages was found to correlate with the suppression of nitrite accumulation in the culture supernatants. Together, these results implicate the L-arginine-dependent production of nitric oxide in T cruzi killing by activated macrophages. We also tested the ability of interleukin(IL)-10 and transforming growth factor (TGF)-beta, to block regulation of T cruzi growth in this system. Both IL-10 and TGF-beta inhibited anti-parasite function by IFN-gamma-activated macrophages, with an optimal dose of 100 units/ml and 0.5 ng/ml, respectively. Moreover, when used in combination, suboptimal doses of IL-10 and TGF-beta were found to produce a synergistic inhibitory effect in the regulation of T cruzi growth. The ability of IL-10 and TGF-beta to suppress microbicidal function was also positively correlated with inhibition of nitrite generation in macrophage culture supernatants. These results predict an in vivo role for IL-10 and TGF-beta in promoting parasite survival in the face of the host cell-mediated immune response.