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tuberculosis has been postulated [ 29, 30?and?31]. Here we sought to determine the possible effect of MtrA overexpression on Mtb sensitivity to cathepsin G. After 24 h incubation of investigated Mtb strains: Rv-wt, Rv-78, Rv-129, and M. bovis BCG with cathepsin G at the final concentrations: 10��g/ml, 50��g/ml, and 100��g/ml, bacteria were plated onto Middlebrook 7H10 solid medium and CFU values were calculated for each strain. The results are shown in Fig. 2. All tested mycobacterial strains, regardless of the used cathepsin G concentrations, showed growth at similar levels, located in the range of 5.263�C6.327 (log10 CFU/ml). Since the survival of the tested strains after 24h did not differ significantly from that in control samples, the mycobacterial strains were subjected to prolonged incubation (72 hours and 120 hours) with cathepsin G at the concentration Selumetinib mouse of 100��g/ml. All tested strains reached a similar growth level, in the range of 4.465�C5.240 log10 CFU/ml. The Bortezomib mw obtained values for control samples were also similar to each other (the lowest value was 4.543 and the highest 5.409) and did not differ significantly from the corresponding tested samples with cathepsin G, with the exception of the M. bovis BCG sample. After 5 days of culturing M. bovis BCG with 100��g/ml cathepsin G, a statistically significant (pDabigatran Moreover, it has been reported that macrophage Mtb infection could lead to a decrease in the mRNA expression of cathepsin G [ 34]. This suggests that such decrease may be a mechanism that is responsible for mycobacterial survival in the hostile environment inside the phagocytes. Thus we decided to examine the possible influence of tested mycobacterial strains on the expression of genes encoding iNOS and cathepsin G in the infected mononuclear phagocytes. The expression of genes for inducible nitric oxide synthase (iNOS) and cathepsin G in THP-1 cells and isolated from blood monocytes stimulated with studied mycobacterial strains are shown in Fig. 3, and the ��-actin housekeeping gene is shown as a positive control. After electrophoresis and staining with ethidium bromide, visible bands indicated the expression of genes for ?-actin and cathepsin G. In THP-1 cells the cathepsin G gene expression remained unchanged regardless of whether the tested cells were stimulated with different mycobacteria strains or they were not stimulated (analysis with use of Kruskal-Wallis test; H=0.
