Experimental study on the mechanisms of fusion between heterotypic Leishmania spp. parasitophorous vacuoles

• Grant number: 10/19335-4
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): June 01, 2011
• Effective date (End): November 30, 2014
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Renato Arruda Mortara
• Grantee: Fernando Roberto Oliveira Real
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 11/51475-3 - Molecular and cellular biology of the parasitism by Trypanosoma cruzi, AP.TEM

Abstract

Classic or modified phagocytosis is the main mechanism of entrance into host cells displayed by several intracellular pathogens. Phagosomes constitute the first, and sometimes, the last stop for the pathogen; they modulate the initial or final pathogens micro-environment, allow pathogen differentiation into different evolutive forms, controll the passage of nutrients and enzimatic substrates (by the presence of channels and by fusion with other host cell vesicles), regulate the antigen presentation by infected cells or the entrance/activity of chemotherapics. Each phagosome is unique, developed from a conjoint of host cell signals triggered by the recognition of the particle/pathogen and from a series of fusion/fission events between phagosome and other selected vesicles. Once internalized by host cells, some pathogens escape from phagosomes and lodge in the citosol. Other are capable of interfere in the maturation of phagosomes, leading to the formation of specialized phagosomes (called parasitophorous vacuoles), customized, displaying selective fusogenicity with host cell vesicles from endocytic and secretory pathways. Parasite of Leishmania genus are examples of pathogens lodged within parasitophorous vacuoles (PVs) during all their intracellular lifecycle inside mammalian host cells. The morphological and biochemical diversity of Leishmania vacuoles is poorly understood. On the other hand, parasites from Trypanosoma cruzi species only transiently live within PVs during their intracellular lifecycle, escaping to host cell citosol (a process which molecular mechanisms are not established). By coinfecting cells in vitro with these two parasites, we investigate the possibility of fusion between Leishmania and T. cruzi PVs and the consequences for both parasites of this intravacuolar cohabitation (chimeric PV). As models of host cells, we will use cell lines transfected with interference RNA for endossomal and lisossomal proteins (LAMPs and Rabs), proteins involved in membrane fusion (SNAREs) and proteins involved in vacuolar acidification (V-ATPases). The participation of these molecules in the transference of T. cruzi to Leishmania PVs, in the differentiation of T. cruzi within chimeric PVs and in the escape of T. cruzi from the chimeric PVs to the citosol will be investigated.

Scientific publications

(5)

(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)

GONCALVES, VIVIANE MARTINELLI; D'ALMEIDA, VANIA; MUELLER, KAREN BARBOSA; REAL, FERNANDO; MORTARA, RENATO ARRUDA. Lysosomal integral membrane protein 2 (LIMP-2) restricts the invasion of Trypanosoma cruzi extracellular amastigotes through the activity of the lysosomal enzyme beta-glucocerebrosidase. Microbes and Infection, v. 16, n. 3, p. 253-260, MAR 2014. (10/19335-4)

PESSOA, CARINA CARRARO; FERREIRA, EDEN RAMALHO; BAYER-SANTOS, ETHEL; RABINOVITCH, MICHEL; MORTARA, RENATO ARRUDA; REAL, FERNANDO. Trypanosoma cruzi Differentiates and Multiplies within Chimeric Parasitophorous Vacuoles in Macrophages Coinfected with Leishmania amazonensis. Infection and Immunity, v. 84, n. 5, p. 1603-1614, MAY 2016. (11/51475-3, 10/19335-4)

REAL, FERNANDO; VIDAL, RAMON OLIVEIRA; CARAZZOLLE, MARCELO FALSARELLA; COSTA MONDEGO, JORGE MAURICIO; LACERDA COSTA, GUSTAVO GILSON; HERAI, ROBERTO HIROCHI; WUERTELE, MARTIN; DE CARVALHO, LUCAS MIGUEL; FERREIRA, RENATA CARMONA E; MORTARA, RENATO ARRUDA; et al. The Genome Sequence of Leishmania (Leishmania) amazonensis: Functional Annotation and Extended Analysis of Gene Models. DNA Research, v. 20, n. 6, p. 567-581, DEC 2013. (10/19335-4)

FLORENTINO, P. T. V.; REAL, F.; BONFIM-MELO, A.; ORIKAZA, C. M.; FERREIRA, E. R.; PESSOA, C. C.; LIMA, B. R.; SASSO, G. R. S.; MORTARA, R. A.. An Historical Perspective on How Advances in Microscopic Imaging Contributed to Understanding the Leishmania Spp. and Trypanosoma cruzi Host-Parasite Relationship. BIOMED RESEARCH INTERNATIONAL, 2014. (14/21338-2, 10/19335-4, 13/05385-8, 12/23150-5)

REAL, FERNANDO; VERAS FLORENTINO, PILAR TAVARES; REIS, LUIZA CAMPOS; RAMOS-SANCHEZ, EDUARDO M.; TAVARES VERAS, PATRICIA SAMPAIO; GOTO, HIRO; MORTARA, RENATO ARRUDA. Cell-to-cell transfer of Leishmania amazonensis amastigotes is mediated by immunomodulatory LAMP-rich parasitophorous extrusions. Cellular Microbiology, v. 16, n. 10, p. 1549-1564, OCT 2014. (10/19335-4)