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Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles
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 livin...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4524717/ https://www.ncbi.nlm.nih.gov/pubmed/26241746 http://dx.doi.org/10.1371/journal.pone.0134925 |
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author | Guedes, Carlos E. S. Lima, Jose G. B. Helfer, Emmanuèle Veras, Patricia S. T. Viallat, Annie |
author_facet | Guedes, Carlos E. S. Lima, Jose G. B. Helfer, Emmanuèle Veras, Patricia S. T. Viallat, Annie |
author_sort | Guedes, Carlos E. S. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4524717 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-45247172015-08-06 Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles Guedes, Carlos E. S. Lima, Jose G. B. Helfer, Emmanuèle Veras, Patricia S. T. Viallat, Annie PLoS One Research Article 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. Public Library of Science 2015-08-04 /pmc/articles/PMC4524717/ /pubmed/26241746 http://dx.doi.org/10.1371/journal.pone.0134925 Text en © 2015 Guedes et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Guedes, Carlos E. S. Lima, Jose G. B. Helfer, Emmanuèle Veras, Patricia S. T. Viallat, Annie Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles |
title | Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles |
title_full | Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles |
title_fullStr | Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles |
title_full_unstemmed | Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles |
title_short | Encapsulation of Living Leishmania Promastigotes in Artificial Lipid Vacuoles |
title_sort | encapsulation of living leishmania promastigotes in artificial lipid vacuoles |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4524717/ https://www.ncbi.nlm.nih.gov/pubmed/26241746 http://dx.doi.org/10.1371/journal.pone.0134925 |
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