AI Article Synopsis
- The chapter presents a viability assay for Leishmania infantum that utilizes bioluminescence for detection.
- It involves a reporter cell line that expresses luciferase and murine macrophages for infection, chosen for their practicality and ethical considerations.
- The assay is designed for screening bioactive compounds in a 96-well microplate format, combining simplicity with effective detection of drug-like molecules.
Article 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.
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