These genes had already been reported to be differentially expressed by peritoneal
macrophages infected with P. brasiliensis . In our experiments, trl2, cd14, Il-1β, nfkb, and tnf-α genes, which play an Pifithrin-�� ic50 important role in the host innate response, were down-regulated during P. brasiliensis-MH-S cell interaction in the presence of pulmonary surfactant or alexidine dihydrochloride compared to the control (Figure 3). In contrast, the main up-regulated genes were those encoding the membrane-related protein CLEC 2 (clec2) – a mannose-type receptor, important for more effective phagocytic capacity  – and the pro-inflammatory inhibitor (nkrf), presenting fold-changes of 8.0 and 9.8 respectively, in cultures exposed to the pulmonary surfactant Oligomycin A solubility dmso (Figure 3). Figure 3 Real-Time RT-PCR. Analysis of the transcript
level of macrophage genes related to phagocytosis (clec2, trl2, and cd14) and inflammation (nkrf, nfkb, tnf-α, and il-1β). The Selleckchem GDC0449 assay was carried out in triplicate (mean ± SEM, with *significance assumed in the range of P < 0:05); **Significantly different from controls: P < 0.001 by the paired 2-tailed Student's t-test. NFkB is a key transcription factor involved in TLR-mediated Liothyronine Sodium innate immunity and together with its repressor Nkrf is an important regulator of the inflammatory process, a powerful protective mechanism coordinated and controlled by cytokines and chemokines. Our data showed an up-regulation of the nkrf gene in the presence of the pulmonary surfactant, suggesting a possible modulation of the
innate immune response under conditions of increased PLB activity. Cytokine production by MH-S cells during host-pathogen interaction In order to verify the pattern of MH-S cell activation, the levels of the cytokines interleukin-10 (IL-10), IL-12, and tumor necrosis factor-α (TNF-α) were determined. When compared to the control, the MH-S cells treated with alexidine produced higher levels of IL-12 and TNF-α and lower levels of IL-10. However, no significant difference between the control group and the group treated with surfactant was observed (Figure 4). Figure 4 Amount of cytokines and tumor necrosis factor-α released by alveolar macrophage (MH-S) cells infected with Paracoccidioides brasiliensis. The assay was carried out in triplicate (mean ± SEM); ns = non-significantly and *significantly different from controls: P < 0.05 by the paired 2-tailed Student’s t-test.