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A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
Published on: January 17, 2014
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Phenotypic assays for Mycobacterium tuberculosis infection
Ok-Ryul Song1, Nathalie Deboosere1, Vincent Delorme1,2
1University of Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019 - UMR 8204 - CIIL - Centre d'Infection et d'Immunité de Lille, F-59000, France.
Summary
Developing new tuberculosis drugs is crucial due to rising drug resistance. This study presents novel assays to monitor Mycobacterium tuberculosis within host cells, aiding drug discovery for this global threat.
Area of Science:
- Microbiology
- Cell Biology
- Drug Discovery
Background:
- Tuberculosis (TB) remains a significant global health issue, with over a million deaths annually.
- Increasing drug resistance in Mycobacterium tuberculosis necessitates novel therapeutic strategies.
- Traditional drug screening methods have yielded limited new anti-TB drugs, highlighting the need for alternative approaches like host-directed therapy.
Purpose of the Study:
- To develop and validate fluorescence-based, high-throughput/high-content (HT/CS) phenotypic assays for monitoring intracellular Mycobacterium tuberculosis.
- To enable drug discovery strategies targeting intracellular pathogens within host macrophages.
- To establish a population-based, multi-parametric analysis pipeline for quantifying host-pathogen interactions.
Main Methods:
- Development of fluorescence-based HT/CS phenotypic assays to track intracellular M. tuberculosis replication and trafficking.
- Implementation of a multi-parametric analysis pipeline to quantify cellular events.
- Monitoring phagosomal maturation (acidification, permeabilization), zinc poisoning, and lipid body accumulation within infected host cells.
Main Results:
- The developed assays successfully monitor the intracellular fate of M. tuberculosis.
- The analysis pipeline quantifies key host-pathogen interplay dynamics, including phagosomal events.
- This approach provides a robust method for assessing drug efficacy against intracellular bacteria.
Conclusions:
- Novel HT/CS phenotypic assays and a multi-parametric analysis pipeline can effectively monitor intracellular Mycobacterium tuberculosis.
- This platform facilitates drug discovery for TB and potentially other intracellular pathogens by quantifying host-pathogen interactions.
- The developed methodology supports the advancement of host-directed therapies against tuberculosis.
Keywords:
automated image-analysiscell-based assayconfocal imagingdrug discoverygenetic screeningmycobacterium tuberculosisMore Related Videos
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