title: Changes in the proteome and infectivity of Leishmania infantum induced by in vitro exposure to a nitric oxide donor.
abstract: Leishmania species are protozoan parasites that exhibit an intracellular amastigote form within mammalian macrophages and an extracellular promastigote form inside the sandfly vector. The generation of nitric oxide (NO) upon activation of macrophages is surely the principal killing effector of intracellular amastigotes but little is known about the potential action of NO against the promastigote phase during its multiplication inside the digestive tract of the sandfly vector. Therefore, we have approached this issue by using an in vitro model to study the effect of an NO donor, 3-morpholinosydnonimine (SIN-1), on the proteome and infectivity of promastigotes of Leishmania infantum. Exposure of promastigotes to SIN-1 during its logarithmic growth phase caused a dramatic effect on parasite protein expression and viability, consequently killing about 60-70% of the promastigotes. The significant changes in the proteome included the over-expression of enolase, peroxidoxin precursors, and heat-shock protein 70 (HSP70), under-expression of 20S proteasome alpha 5 unit, and phosphomannomutase and induced expression of 3-hydroxy-3-methyglutaryl-CoA (HMG-CoA) synthase and prostaglandine f2-alpha (PGD2) synthase. Interestingly, promastigotes that resisted treatment showed enhanced infectivity to J774 macrophages in comparison to the controls. This finding together with the appearance of the PGD2S and an over-expression of HSP70 isoforms in treated promastigotes led us to speculate the existence of NO-mediated programmed cell death (PCD) events as a potential mechanism of population regulation and selection of properly infecting forms that predominantly operate on the promastigote stage.
authors.affiliation: Departamento de Parasitología, Universidad Complutense, Ciudad Universitaria, Madrid, Spain.
authors.forename: Maria Auxiliadora
authors.initials: MA
authors.lastname: Dea-Ayuela
authors.forename: Lara
authors.initials: L
authors.lastname: Ordoñez-Gutierrez
authors.forename: Francisco
authors.initials: F
authors.lastname: Bolás-Fernández
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D002460:Cell Line; D002470:Cell Survival; D005786:Gene Expression Regulation / Q000187:drug effects; D018314:Leishmania infantum / Q000737:chemistry* / Q000187:drug effects*; D008264:Macrophages / Q000469:parasitology; D051379:Mice; D008981:Molsidomine / Q000031:analogs & derivatives* / Q000494:pharmacology; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D020543:Proteome / Q000032:analysis*; D015800:Protozoan Proteins / Q000032:analysis*
chemical_list: D000981:Antiprotozoal Agents; D020030:Nitric Oxide Donors; D020543:Proteome; D015800:Protozoan Proteins; C002385:linsidomine; D008981:Molsidomine