The intracellular infection by protozoa of the genus Leishmania involves the formation of parasitophorous vacuoles (PV) in which the parasite develops and multiplies within macrophages. Leishmania (Leishmania) mexicana and Leishmania (Leishmania) amazonensis multiply in spacious PVs that contain more than one amastigote, while most of the species studied as L. (L.) major and L. (L.) donovani form PVs juxtaposed to membrane of amastigotes. Leishmania PVs present in their membrane MHC II molecules and they have acidic pH, maintained by vacuolar ATPase proton (v-ATPase). Some isoforms of V-ATPases are more expressed in macrophages infected with L. (L.) amazonensis compared with uninfected macrophages. This is the case of the d2 subunit (ATP6V0d2) whose function or participate in intracellular infection by this parasite is unknown. Preliminary results showed that the growth vacuolar in the infection by L. (L.) amazonensis in silenced cells to the d2 isoform was impaired and cholesterol expression in these same macrophages was changed. In this project, we propose the use of RAW 264.7 macrophages silenced for d2 isoform in order to investigate further the role of v-ATPase in the biogenesis of the PVs formed by L. (L.) amazonensis, L. (L.) mexicana and Coxiella burnetii. In addition, considering the hypothetical interference of spacious VPs in presenting antigens of intracellular pathogens, we will investigate the volumetric reduction of vacuoles in macrophages silenced for ATP6V0d2 can interfere with antigen presentation via MHC class II and / or I by these macrophages.
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