Transcriptome of human monocyte-derived macrophages infected with live-Mycobacterium leprae. 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. Overall design: 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.