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. 2022:2524:127-147.
doi: 10.1007/978-1-0716-2453-1_10.

A Simple Bioluminescent Assay for the Screening of Cytotoxic Molecules Against the Intracellular Form of Leishmania infantum

Diego Benítez  1 , Andrea Medeiros  2   3 , Cristina Quiroga  2 , Marcelo A Comini  4
Affiliations

A Simple Bioluminescent Assay for the Screening of Cytotoxic Molecules Against the Intracellular Form of Leishmania infantum

Diego Benítez et al. Methods Mol Biol. 2022.

Abstract

This chapter describes a viability assay for the intracellular (amastigote) and clinically relevant form of Leishmania infantum that is based on the detection of bioluminescence (BL) signal. The assay uses a reporter cell line of L. infantum that expresses constitutively a redshifted luciferase from Photinus pyralis and murine macrophages (cell line J774.A1) as host cells for infection. The host cell line was selected because it is a differentiated cell line, easy to manipulate in vitro, and advantageous for ethical reasons. This chapter introduces an assay designed for the screening of bioactive compounds/molecules employing a 96-well microplate and a 24 h treatment. The assay setup shows excellent balance between simplicity (cell culture manipulation/infection and timing) and quality parameters, as well as potential to detect drug-like molecules acting in a fast and cytotoxic manner.

Keywords: Drug discovery; Leishmania infantum; Redshifted luciferase; Viability assay; Visceral leishmaniasis.

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