title: Nitric oxide participation in the fungicidal mechanism of gamma interferon-activated murine macrophages against Paracoccidioides brasiliensis conidia.
abstract: Paracoccidioidomycosis, a systemic mycosis restricted to Latin America and produced by the dimorphic fungus Paracoccidioides brasiliensis, is probably acquired by inhalation of conidia produced by the mycelial form. The macrophage (Mphi) represents the major cell defense against this pathogen; when activated with gamma interferon (IFN-gamma), murine Mphis kill the fungus by an oxygen-independent mechanism. Our goal was to determine the role of nitric oxide in the fungicidal effect of Mphis on P. brasiliensis conidia. The results revealed that IFN-gamma-activated murine Mphis inhibited the conidium-to-yeast transformation process in a dose-dependent manner; maximal inhibition was observed in Mphis activated with 50 U/ml and incubated for 96 h at 37 degrees C. When Mphis were activated with 150 to 200 U of cytokine per ml, the number of CFU was 70% lower than in nonactivated controls, indicating that there was a fungicidal effect. The inhibitory effect was reversed by the addition of anti-IFN-gamma monoclonal antibodies. Activation by IFN-gamma also enhanced Mphi nitric oxide production, as revealed by increasing NO(2) values (8 +/- 3 microM in nonactivated Mphis versus 43 +/- 13 microM in activated Mphis). The neutralization of IFN-gamma also reversed nitric oxide production at basal levels (8 +/- 5 microM). Additionally, we found that there was a significant inverse correlation (r = -0.8975) between NO(2)(-) concentration and transformation of P. brasiliensis conidia. Additionally, treatment with any of the three different nitric oxide inhibitors used (arginase, N(G)-monomethyl-L-arginine, and aminoguanidine), reverted the inhibition of the transformation process with 40 to 70% of intracellular yeast and significantly reduced nitric oxide production. These results show that IFN-gamma-activated murine Mphis kill P. brasiliensis conidia through the L-arginine-nitric oxide pathway.
authors.affiliation: Medical and Experimental Mycology Group, Corporación para Investigaciones Biológicas, Medellín, Colombia.
authors.forename: A
authors.initials: A
authors.lastname: Gonzalez
authors.forename: W
authors.initials: W
authors.lastname: de Gregori
authors.forename: D
authors.initials: D
authors.lastname: Velez
authors.forename: A
authors.initials: A
authors.lastname: Restrepo
authors.forename: L E
authors.initials: LE
authors.lastname: Cano
mesh_terms: D000818:Animals; D001119:Arginase / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006146:Guanidines / Q000494:pharmacology; D007371:Interferon-gamma / Q000276:immunology*; D007424:Intracellular Fluid / Q000382:microbiology; D008262:Macrophage Activation / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009569:Nitric Oxide / Q000096:biosynthesis / Q000276:immunology*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D010228:Paracoccidioides / Q000276:immunology*; D019323:omega-N-Methylarginine / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D006146:Guanidines; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D007371:Interferon-gamma; D019001:Nitric Oxide Synthase; D001119:Arginase; C004479:pimagedine