Development of an Intracellular Screen for New Compounds Able To Inhibit Mycobacterium tuberculosis Growth in Human Macrophages.
Here we describe the development and validation of an intracellular high-throughput screening assay for finding new antituberculosis compounds active in human macrophages. The assay consists of a luciferase-based primary identification assay, followed by a green fluorescent protein-based secondary profiling assay. Standard tuberculosis drugs and 158 previously recognized active antimycobacterial compounds were used to evaluate assay robustness. Data show that the assay developed is a short and valuable tool for the discovery of new antimycobacterial compounds.
Department of Medicine, Division of Infectious Diseases, Faculty of Medicine, University of British Columbia, Vancouver, British Columbia, Canada Diseases of the Developing World, GSK, Tres Cantos, Madrid, Spain.
Flavia
F
Sorrentino
Diseases of the Developing World, GSK, Tres Cantos, Madrid, Spain.
Ruben
R
Gonzalez del Rio
Department of Medicine, Division of Infectious Diseases, Faculty of Medicine, University of British Columbia, Vancouver, British Columbia, Canada.
Xingji
X
Zheng
Diseases of the Developing World, GSK, Tres Cantos, Madrid, Spain.
Jesus
J
Presa Matilla
Diseases of the Developing World, GSK, Tres Cantos, Madrid, Spain.
Pedro
P
Torres Gomez
Diseases of the Developing World, GSK, Tres Cantos, Madrid, Spain.
Maria
M
Martinez Hoyos
Diseases of the Developing World, GSK, Tres Cantos, Madrid, Spain.
Maria Esther
ME
Perez Herran
Diseases of the Developing World, GSK, Tres Cantos, Madrid, Spain.
Alfonso
A
Mendoza Losana
Department of Medicine, Division of Infectious Diseases, Faculty of Medicine, University of British Columbia, Vancouver, British Columbia, Canada yossi@mail.ubc.ca.
Yossef
Y
Av-Gay
D000995:Antitubercular Agents / Q000737:chemistry / Q000494:pharmacology*; D002460:Cell Line; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects* / Q000382:microbiology; D008826:Microbial Sensitivity Tests; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000254:growth & development; D013329:Structure-Activity Relationship
D000995:Antitubercular Agents; D049452:Green Fluorescent Proteins; D008156:Luciferases