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Investigation of the role of vacuolar ATPase of d2 subunit ( ATP6V0d2 ) in the biogenese of parasitophorous vacuoles formed by Leishmania (Leishmania) amazonensis in RAW 264.7 macrophages

Grant number: 15/14205-9
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): November 01, 2015
Effective date (End): April 30, 2019
Field of knowledge:Biological Sciences - Parasitology - Protozoology of Parasites
Principal Investigator:Renato Arruda Mortara
Grantee:Carina Carraro Pessoa
Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

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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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
PESSOA, CARINA CARRARO; REIS, LUIZA CAMPOS; RAMOS-SANCHEZ, EDUARDO MILTON; ORIKAZA, CRISTINA MARY; CORTEZ, CRISTIAN; DE CASTRO LEVATTI, ERICA VALADARES; BENITES BADARO, ANA CAROLINA; DA SILVA YAMAMOTO, JOYCE UMBELINO; D'ALMEIDA, VANIA; GOTO, HIRO; et al. ATP6V(0)d2 controls Leishmania parasitophorous vacuole biogenesis via cholesterol homeostasis. PLOS PATHOGENS, v. 15, n. 6, . (15/14205-9, 16/15000-4)

Please report errors in scientific publications list by writing to: cdi@fapesp.br.