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

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) assa...

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Bibliographic Details
Published in:PloS one 2018-08, Vol.13 (8), p.e0201678
Main Authors: Touquet, Bastien, Pelissier, Léonie, Cavailles, Pierre, Yi, Wei, Bellini, Valeria, Mercier, Corinne, Cesbron-Delauw, Marie-France, Boumendjel, Ahcène, Aldebert, Delphine
Format: Article
Language:English
Subjects:
Animals
Assaying
Automation
Biology and Life Sciences
Care and treatment
Cell proliferation
Cells, Cultured
Fibroblasts - cytology
Fibroblasts - parasitology
Fluorescence
Fluorescence microscopy
Fluorescent Dyes - metabolism
Fluorescent indicators
Foreskin - cytology
Foreskin - parasitology
Humans
Image analysis
Image processing
Infections
Inhibition
Life Sciences
Male
Medicine and Health Sciences
Microbiology and Parasitology
Microscopy, Fluorescence
Parasites
Protozoa
Risk factors
RNA polymerase
Software
Thiosemicarbazones - pharmacology
Toxicity
Toxoplasma
Toxoplasma - drug effects
Toxoplasma - growth & development
Toxoplasma gondii
Toxoplasmosis
Toxoplasmosis - diagnostic imaging
Toxoplasmosis - metabolism
Uracil
Uracil - metabolism
Womens health
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0201678