High-content imaging assay to evaluate Toxoplasma gondii infection and proliferation: A multiparametric assay to screen new compounds.

Autor: Touquet B; Institute for Advanced Biosciences, Team Membrane Dynamics of Parasite-Host Cell Interactions, CNRS UMR 5309, INSERM U1209, Université Grenoble Alpes, Grenoble, France., Pelissier L; Unité Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Equipe TheREx, CNRS UMR 5525, Université Grenoble Alpes, Grenoble, France.; Département de Pharmacochimie Moléculaire, CNRS UMR 5063, Université Grenoble Alpes, Grenoble, France., Cavailles P; Unité Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Equipe BNI, CNRS UMR 5525, Université Grenoble Alpes, Grenoble, France., Yi W; Département de Pharmacochimie Moléculaire, CNRS UMR 5063, Université Grenoble Alpes, Grenoble, France., Bellini V; Institute for Advanced Biosciences, Team Host-Pathogen Interactions and Immunity to Infection, CNRS UMR 5309, INSERM U1209, Université Grenoble Alpes, Grenoble, France., Mercier C; Unité Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Equipe GEM, CNRS UMR 5525, Université Grenoble Alpes, Grenoble, France., Cesbron-Delauw MF; Unité Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Equipe BNI, CNRS UMR 5525, Université Grenoble Alpes, Grenoble, France., Boumendjel A; Département de Pharmacochimie Moléculaire, CNRS UMR 5063, Université Grenoble Alpes, Grenoble, France., Aldebert D; Unité Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications, Equipe TheREx, CNRS UMR 5525, Université Grenoble Alpes, Grenoble, France.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2018 Aug 29; Vol. 13 (8), pp. e0201678. Date of Electronic Publication: 2018 Aug 29 (Print Publication: 2018).
DOI: 10.1371/journal.pone.0201678
Abstrakt: Toxoplasma gondii is an intracellular protozoan parasite widely distributed in animals and humans. Infection of host cells and parasite proliferation are essential steps in Toxoplasma pathology. The objective of this study was to develop and validate a novel automatic High Content Imaging (HCI) assay to study T. gondii infection and proliferation. We tested various fluorescent markers and strategies of image analysis to obtain an automated method providing results comparable to those from gold standard infection and proliferation assays. No significant difference was observed between the results obtained from the HCI assay and the standard assays (manual fluorescence microscopy and incorporation of [3H]-uracil). We developed here a robust and time-saving assay. This automated technology was then used to screen a library of compounds belonging to four classes of either natural compounds or synthetic derivatives. Inhibition of parasite proliferation and host cell toxicity were measured in the same assay and led to the identification of one hit, a thiosemicarbazone that allows important inhibition of Toxoplasma proliferation while being relatively safe for the host cells.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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