Figure 3

The activation profiles of macrophages infected with pathogenic mycobacteria. BMDM were infected with the studied mycobacterial strains at a MOI of 5:1, washed and incubated for an additional 48 h. The cells left untreated and cells stimulated with LPS for 48 h were used as a negative and positive controls of proinflammatory activation, respectively. To evaluate markers of M1-type activation (A), the culture supernatants of infected cultures were harvested and tested for TNF-α, IL-6, MCP-1, MIP-2 and IL-12 by Bioplex test, and for NO production by Griess reaction. Assays were completed with duplicate samples, and results are expressed as a mean of three independent experiments. Lines over bars indicate the isolates for which the induced cytokine production differed significantly from that induced by H37Rv (*p < 0.05; ***p < 0.001). To evaluate markers of M2-type activation, secretion of IL-10 was quantified by Bioplex assay (B), and expression of Arginase 1 and MR/CD206 in the adhered cells was tested by Western blotting (C). Lower panel, quantification of the protein levels by densitometric analysis of immunoreactive bands.