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Immunology Program, Departments of
Internal Medicine and
Microbiology, and
Program in Free Radical and Radiation Biology, University of Iowa, and
¶ Veterans Affairs Medical Center, Iowa City, IA 52242; and
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Universidade Federal do Rio Grande do Norte, Natal, Brazil
Leishmania chagasi, the cause of South American
visceral leishmaniasis, must survive antimicrobial responses of host
macrophages to establish infection. Macrophage oxidative responses have
been shown to diminish in the presence of intracellular leishmania.
However, using electron spin resonance we demonstrated that murine and
human macrophages produce O2- during
phagocytosis of opsonized promastigotes. Addition of the
O2- scavenger
4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl to
cultures resulted in increased infection, suggesting that
O2- enhances macrophage
leishmanicidal activity. The importance of NO· produced by
inducible NO synthase (iNOS) in controlling murine leishmaniasis is
established, but its role in human macrophages has been debated. We
detected NO· in supernatants from murine, but not human,
macrophages infected with L. chagasi. Nonetheless, the
iNOS inhibitor
NG-monomethyl-L-arginine
inhibited IFN-
-mediated intracellular killing by both murine and
human macrophages. According to RNase protection assay and
immunohistochemistry, iNOS mRNA and protein were expressed at higher
levels in bone marrow of patients with visceral leishmaniasis than in
controls. The iNOS protein also increased upon infection of human
macrophages with L. chagasi promastigotes in vitro in
the presence of IFN-. These data suggest that
O2- and NO· each contribute to
intracellular killing of L. chagasi in human and murine
macrophages.
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