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https://www.arca.fiocruz.br/handle/icict/13430
ENCAPSULATION OF LIVING LEISHMANIA PROMASTIGOTES IN ARTIFICIAL LIPID VACUOLES
Autor(es)
Afiliação
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil
Aix-Marseille Université. CNRS. Marseille, France
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil
Aix-Marseille Université. CNRS. Marseille, France
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil
Aix-Marseille Université. CNRS. Marseille, France
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório de Patologia e Biointervenção. Salvador, BA, Brasil
Aix-Marseille Université. CNRS. Marseille, France
Resumo em Inglês
After phagocytosis by mammalian macrophages, promastigote forms of Leishmania parasites
settle inside intracellular parasitophorous vacuoles (PVs) in which they transform into
amastigote forms and replicate. Here, using a variant of the ‘inverted emulsion’ method, we
succeeded in encapsulating living L. amazonensis parasites in giant artificial liposomes that
serve as model PVs. We were able to control the size of liposomes, the pH and the composition
of their internal volume, and the number of internalized parasites per liposome. L.
amazonensis promastigotes encapsulated in liposomes filled with RPMI-Dextran solution at
pH 7.5 or 6.5 survived up to 96 h at 24°C. At 37°C and pH 5.5, parasites survived 48h. This
method paves the way to identifying certain effectors secreted by the parasite and to unraveling
specific mechanisms of fusion between the PV and intracellular vesicles of the host
cell. This method will also facilitate the study of the temporal evolution of biophysical properties
of the PV during its maturation.
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