title: Tissue expression of inducible nitric oxide synthase is closely associated with resistance to Leishmania major.
abstract: Previous studies with inhibitors of inducible nitric oxide synthase (iNOS) suggested that high-output production of nitric oxide (NO) is an important antimicrobial effector pathway in vitro and in vivo. Here, we investigated the tissue expression of iNOS in mice after infection with Leishmania major. Immunohistochemical staining with an iNOS-specific antiserum revealed that in the cutaneous lesion and draining lymph nodes (LN) of clinically resistant mice (C57BL/6), iNOS protein is found earlier during infection and in significantly higher amounts than in the nonhealing BALB/c strain. Similar differences were seen on the mRNA level as quantitated by competitive polymerase chain reaction. Anti-CD4 treatment of BALB/c mice not only induced resistance to disease, but also restored the expression of iNOS in the tissue. In situ, few or no parasites were found in those regions of the skin lesion and the draining LN which were highly positive for iNOS. By double labeling experiments, macrophages were identified as iNOS expressing cells in vivo. In the lesions of BALB/c mice, cells staining positively for transforming growth factor beta (TGF-beta), a potent inhibitor of iNOS in vitro, were strikingly more prominent than in C57BL/6, whereas no such difference was found for interleukin 4 or interferon gamma (IFN-gamma). In vitro, production of NO was approximately threefold higher in C57BL/6 than in BALB/c macrophages after stimulation with IFN-gamma. We conclude that the pronounced expression of iNOS in resistant mice is an important mechanism for the elimination of Leishmania in vivo. The relative lack of iNOS in susceptible mice might be a consequence of macrophage deactivation by TGF-beta and reduced responsiveness to IFN-gamma.
authors.affiliation: Institute of Clinical Microbiology and Immunology, University of Erlangen, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Stenger
authors.forename: H
authors.initials: H
authors.lastname: Thüring
authors.forename: M
authors.initials: M
authors.lastname: Röllinghoff
authors.forename: C
authors.initials: C
authors.lastname: Bogdan
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D005260:Female; D007371:Interferon-gamma / Q000494:pharmacology; D018320:Leishmania major; D016773:Leishmaniasis, Cutaneous / Q000276:immunology*; D008264:Macrophages / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009252:NADPH Dehydrogenase / Q000378:metabolism; D019001:Nitric Oxide Synthase; D012333:RNA, Messenger / Q000032:analysis; D016212:Transforming Growth Factor beta / Q000096:biosynthesis
chemical_list: D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D007371:Interferon-gamma; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D009252:NADPH Dehydrogenase