title: Effect of insulin-like growth factor-I on Leishmania amazonensis promastigote arginase activation and reciprocal inhibition of NOS2 pathway in macrophage in vitro.
abstract: We showed previously that insulin-like growth factor (IGF)-I induces an exacerbation of the lesion development in experimental cutaneous leishmaniasis favouring parasite growth within host macrophages. Here we studied the effect of IGF-I in vitro in BALB/c mouse peritoneal macrophages infected with stationary phase Leishmania amazonensis promastigotes. IGF-I was used to pre-incubate either macrophage or parasite before infection of the macrophages or adding it at the start of the Leishmania-macrophage culture and maintaining it throughout the experimental period. Independent of stimulation protocol, IGF-I induced significantly increased parasite growth within macrophages. Arginase activation considered as a key factor in Leishmania growth was studied, and its expression and activity were increased in Leishmania-infected macrophages but significantly more in infected cells upon IGF-I stimulus, an effect specifically inhibited by NOHA. Arginase known to be present on Leishmania was also studied, and its expression and activity were seen in the absence of any stimulus but significantly increased after 5 min of incubation with IGF-I. In addition, Leishmania was pre-incubated with NOHA for 5 min, washed, then macrophages infected observing a significantly reduced parasite burden in both IGF-I-stimulated and non-stimulated macrophages. Reciprocal decrease in the nitric oxide (NO) level and inhibition of nitric oxide synthase (NOS2) expression were also observed in IGF-I-stimulated infected macrophages. Our data strongly suggest that IGF-I induces preferential expression and activation of Leishmania promastigote arginase, contributes to the alternative activation of macrophages in the context of innate immunity and interferes with NOS pathway in infected macrophages probably as a reciprocal effect.
authors.affiliation: Laboratório de Soroepidemiologia e Imunobiologia, Instituto de Medicina Tropical de São Paulo, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: C M V
authors.initials: CM
authors.lastname: Vendrame
authors.forename: M D T
authors.initials: MD
authors.lastname: Carvalho
authors.forename: F J O
authors.initials: FJ
authors.lastname: Rios
authors.forename: E R
authors.initials: ER
authors.lastname: Manuli
authors.forename: F
authors.initials: F
authors.lastname: Petitto-Assis
authors.forename: H
authors.initials: H
authors.lastname: Goto
mesh_terms: D000818:Animals; D001119:Arginase / Q000378:metabolism*; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000276:immunology; D005260:Female; D006790:Host-Parasite Interactions / Q000276:immunology; D007334:Insulin-Like Growth Factor I / Q000502:physiology*; D007894:Leishmania mexicana / Q000201:enzymology* / Q000254:growth & development / Q000276:immunology; D017737:Macrophages, Peritoneal / Q000201:enzymology* / Q000276:immunology / Q000469:parasitology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors* / Q000502:physiology; D015398:Signal Transduction / Q000276:immunology
chemical_list: D007334:Insulin-Like Growth Factor I; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D001119:Arginase