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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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0001671&type=printable |
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| 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 |
| collection | DOAJ |
| 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 |
| id | doaj-art-a71f7b0e7374429b8ffc953440c372f8 |
| institution | Kabale University |
| issn | 1935-2727 1935-2735 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj-art-a71f7b0e7374429b8ffc953440c372f82025-01-16T05:32:28ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352012-01-0166e167110.1371/journal.pntd.0001671An image-based high-content screening assay for compounds targeting intracellular Leishmania donovani amastigotes in human macrophages.Jair L Siqueira-NetoSeunghyun MoonJiyeon JangGyongseon YangChangbok LeeHong Kee MoonEric ChatelainAuguste GenovesioJonathan CechettoLucio H Freitas-JuniorLeishmaniasis 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.https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0001671&type=printable |
| spellingShingle | 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. PLoS Neglected Tropical Diseases |
| title | 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_short | 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 |
| url | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0001671&type=printable |
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