An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages.
Leishmaniasis is a tropical disease threatening 350 million people from endemic regions. The available drugs for treatment are inadequate, with limitations such as serious side effects, parasite resistance or high cost. Driven by this need for new drugs, we developed a high-content, high-throughput...
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ftdoajarticles:oai:doaj.org/article:a71f7b0e7374429b8ffc953440c372f8 2023-05-15T15:08:54+02:00 An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages. Jair L Siqueira-Neto Seunghyun Moon Jiyeon Jang Gyongseon Yang Changbok Lee Hong Kee Moon Eric Chatelain Auguste Genovesio Jonathan Cechetto Lucio H Freitas-Junior 2012-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0001671 https://doaj.org/article/a71f7b0e7374429b8ffc953440c372f8 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3373640?pdf=render https://doaj.org/toc/1935-2735 1935-2735 doi:10.1371/journal.pntd.0001671 https://doaj.org/article/a71f7b0e7374429b8ffc953440c372f8 PLoS Neglected Tropical Diseases, Vol 6, Iss 6, p e1671 (2012) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2012 ftdoajarticles https://doi.org/10.1371/journal.pntd.0001671 2022-12-31T13:30:15Z Leishmaniasis is a tropical disease threatening 350 million people from endemic regions. The available drugs for treatment are inadequate, with limitations such as serious side effects, parasite resistance or high cost. Driven by this need for new drugs, we developed a high-content, high-throughput image-based screening assay targeting the intracellular amastigote stage of different species of Leishmania in infected human macrophages. The in vitro infection protocol was adapted to a 384-well-plate format, enabling acquisition of a large amount of readouts by automated confocal microscopy. The reading method was based on DNA staining and required the development of a customized algorithm to analyze the images, which enabled the use of non-modified parasites. The automated analysis generated parameters used to quantify compound activity, including infection ratio as well as the number of intracellular amastigote parasites and yielded cytotoxicity information based on the number of host cells. Comparison of this assay with one that used the promastigote form to screen 26,500 compounds showed that 50% of the hits selected against the intracellular amastigote were not selected in the promastigote screening. These data corroborate the idea that the intracellular amastigote form of the parasite is the most appropriate to be used in primary screening assay for Leishmania. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 6 6 e1671 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Jair L Siqueira-Neto Seunghyun Moon Jiyeon Jang Gyongseon Yang Changbok Lee Hong Kee Moon Eric Chatelain Auguste Genovesio Jonathan Cechetto Lucio H Freitas-Junior An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Leishmaniasis is a tropical disease threatening 350 million people from endemic regions. The available drugs for treatment are inadequate, with limitations such as serious side effects, parasite resistance or high cost. Driven by this need for new drugs, we developed a high-content, high-throughput image-based screening assay targeting the intracellular amastigote stage of different species of Leishmania in infected human macrophages. The in vitro infection protocol was adapted to a 384-well-plate format, enabling acquisition of a large amount of readouts by automated confocal microscopy. The reading method was based on DNA staining and required the development of a customized algorithm to analyze the images, which enabled the use of non-modified parasites. The automated analysis generated parameters used to quantify compound activity, including infection ratio as well as the number of intracellular amastigote parasites and yielded cytotoxicity information based on the number of host cells. Comparison of this assay with one that used the promastigote form to screen 26,500 compounds showed that 50% of the hits selected against the intracellular amastigote were not selected in the promastigote screening. These data corroborate the idea that the intracellular amastigote form of the parasite is the most appropriate to be used in primary screening assay for Leishmania. |
format |
Article in Journal/Newspaper |
author |
Jair L Siqueira-Neto Seunghyun Moon Jiyeon Jang Gyongseon Yang Changbok Lee Hong Kee Moon Eric Chatelain Auguste Genovesio Jonathan Cechetto Lucio H Freitas-Junior |
author_facet |
Jair L Siqueira-Neto Seunghyun Moon Jiyeon Jang Gyongseon Yang Changbok Lee Hong Kee Moon Eric Chatelain Auguste Genovesio Jonathan Cechetto Lucio H Freitas-Junior |
author_sort |
Jair L Siqueira-Neto |
title |
An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages. |
title_short |
An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages. |
title_full |
An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages. |
title_fullStr |
An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages. |
title_full_unstemmed |
An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages. |
title_sort |
image-based high-content screening assay for compounds targeting intracellular leishmania donovani amastigotes in human macrophages. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2012 |
url |
https://doi.org/10.1371/journal.pntd.0001671 https://doaj.org/article/a71f7b0e7374429b8ffc953440c372f8 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 6, Iss 6, p e1671 (2012) |
op_relation |
http://europepmc.org/articles/PMC3373640?pdf=render https://doaj.org/toc/1935-2735 1935-2735 doi:10.1371/journal.pntd.0001671 https://doaj.org/article/a71f7b0e7374429b8ffc953440c372f8 |
op_doi |
https://doi.org/10.1371/journal.pntd.0001671 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
6 |
container_issue |
6 |
container_start_page |
e1671 |
_version_ |
1766340178621759488 |