Transcriptome of human monocyte-derived macrophages infected with live-Mycobacterium leprae

To this date, host transcriptome studies in leprosy have focused on Schwann cells, as well as mouse-footpad and skin biopsies. Despite macrophages being the most infected cell types in leprosy lesions, there is no genome-wide experiments with this model. Here, we aimed at identifying host macrophages transcriptional changes induced by live-Mycobacterium leprae infection for 48 hours. Peripheral blood from female and male healthy individuals were collected. PBMCs were isolated by the density gradient method (histopaque 1077) with further monocyte purification using anti-CD14 magnetic beads. Monocytes (95% purity checked by flow cytometry) were seeded (5x10^5 cells/well) in 24-well plates and grown in Iscove’s modified Dubelcco’s medium supplemented with antibiotics and pooled human sera for 6 days at 37 °C in 5% CO2. After macrophage differentiation, cultures were fed by replacing half of medium volume every 3 days. Macrophages were infected with live-M. leprae (Thai-53 strain) with MOI of 10:1 (bacteria:host) for 48 hours.