Functional analysis of oxLDL-treated macrophages.

Primary human macrophages were treated with PBS control, native LDL or oxLDL (1, 10 or 25 μg/ml) overnight. (A) Macrophages were incubated with fluorescent phagocytosis beads at a MOI of 10:1 and subsequently analyzed by flow cytometry. Fluorescence of extracellular beads was quenched with Trypan Blue. (B) Percentage of macrophages with 0, 1 or ≥2 beads internalized beads (n = 6). Data is represented as means with standard deviations. (C) Macrophages were infected with Mtb H37Rv at a MOI of 10:1 for 1 h. Supernatants were harvested at 24 h post-infection and concentrations of TNF-α, IL-6, IL-10 and IL-1β were determined by ELISA (n = 10). Individual donors are depicted as dots with group medians. (D) Macrophages were co-cultured for four days with the HLA-DR2-restricted CD4+ T cell R2F10 at a ratio of 1:4 and 0.1, 1 or 10 μg/ml of its cognate peptide. T cell proliferation was measured by tritium-thymidine incorporation during the last 24 h (n = 3). Data is represented as means with standard deviations. Statistical significance was determined by Wilcoxon signed rank test with post-hoc FDR correction. * = p < 0.05.