title: A High-throughput Compatible Assay to Evaluate Drug Efficacy against Macrophage Passaged Mycobacterium tuberculosis.
abstract: The early drug development process for anti-tuberculosis drugs is hindered by the inefficient translation of compounds with in vitro activity to effectiveness in the clinical setting. This is likely due to a lack of consideration for the physiologically relevant cellular penetration barriers that exist in the infected host. We recently established an alternative infection model that generates large macrophage aggregate structures containing densely packed M. tuberculosis (Mtb) at its core, which was suitable for drug susceptibility testing. This infection model is inexpensive, rapid, and most importantly BSL-2 compatible. Here, we describe the experimental procedures to generate Mtb/macrophage aggregate structures that would produce macrophage-passaged Mtb for drug susceptibility testing. In particular, we demonstrate how this infection system could be directly adapted to the 96-well plate format showing throughput capability for the screening of compound libraries against Mtb. Overall, this assay is a valuable addition to the currently available Mtb drug discovery toolbox due to its simplicity, cost effectiveness, and scalability.
authors.affiliation: Department of Medicine, University of Alabama at Birmingham.
authors.forename: Kaitlyn
authors.initials: K
authors.lastname: Schaaf
authors.affiliation: Department of Medicine, University of Alabama at Birmingham.
authors.forename: Samuel R
authors.initials: SR
authors.lastname: Smith
authors.affiliation: Department of Medicine, University of Alabama at Birmingham.
authors.forename: Virginia
authors.initials: V
authors.lastname: Hayley
authors.affiliation: Department of Medicine, University of Alabama at Birmingham.
authors.forename: Olaf
authors.initials: O
authors.lastname: Kutsch
authors.affiliation: Department of Medicine, University of Alabama at Birmingham; jsun14@uab.edu.
authors.forename: Jim
authors.initials: J
authors.lastname: Sun
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D006801:Humans; D008264:Macrophages / Q000187:drug effects*; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000302:isolation & purification; D014376:Tuberculosis / Q000188:drug therapy / Q000382:microbiology*
chemical_list: D000995:Antitubercular Agents