These data support the notion that inflammasome activation does not occur when the bacteria are confined to intact phagosomes, whereas even the partial disruption of the phagosomal membranes, as executed by ΔpdpC, JAK inhibitor leads to highly significant, but intermediate levels inflammasome-activating cytosolic signaling. This is a slightly modified hypothesis compared to the previously proposed, suggesting that there was a direct correlation between cytosolic location and inflammasome activation [17, 20, 22, 38]. Table 3 IL-1β secretion from F. tularensis-infected BMDM cells Strain IL-1β secretion (pg/ml)a

  5 h 24 h – BDL*** BDL*** LVS 76.3 ± 10.9 497.1 ± 79.0 ΔiglC 39.6b BDL*** ΔpdpC 64.5 ± 27.2 112.1 ± 41.0* ΔpdpC/pdpC 163.2 ± 50.2 506.9 ± 94.3 a F. tularensis-infected, or

uninfected (-) BMDM cells were incubated for 5 or 24 h. The average IL-1β secretion in pg/ml with standard errors of triplicate biological samples from one representative experiment, out of three, is shown. A Student’s t-test was used to determine if the IL-1β secretion was significantly different between LVS infected and mutant infected cells (*: P < 0.05, **: P < 0.01, Trichostatin A ***: P < 0.001). BDL means that the concentration was below the detection limit of the assay (< 31.25 pg/ml). b Only one of the triplicates was above BDL. Discussion F. tularensis is capable of rapid escape from

the phagosome, which is followed by efficient growth within the cytosol of monocytic cells. The molecular mechanisms behind the intracellular life style of the bacterium are not well understood, but have been shown to be dependent on many FPI-encoded genes, of which the most well-studied are the members of the iglABCD operon [16, 28, 37]. Mirabegron Evidence indicates that many of the FPI proteins collectively constitute a T6SS, however, while such systems have been identified in nearly 100 different bacterial species to date, their homologies to the FPI system are weak, indicating that the latter constitutes an evolutionarily distinct group [1, 14, 22]. While the FPI proteins IglA, IglB, PdpB, VgrG, and DotU show modest similarities to common components of T6SSs, the remaining FPI proteins appear to be unique and this makes it MK-8776 mw laborious and tedious to understand their roles and functions. The accumulating evidence indicates that many of them are essential core components and as such critically required and, thereby, their absence leads to a null mutant phenotype characterized by lack of phagosomal escape, no intracellular replication, and avirulence [9]. A majority of the investigated FPI mutants appears to belong to this group but, in contrast, the ΔpdpE mutant exhibits full virulence [17].