Sublethal Doses of Anthrax Lethal Toxin on the Suppression of Macrophage Phagocytosis

BACKGROUND: Lethal toxin (LT), the major virulence factor produced by Bacillus anthracis, has been shown to suppress the immune system, which is beneficial to the establishment of B. anthracis infections. It has been suggested that the suppression of MEK/MAPK signaling pathways of leukocytes contributes to LT-mediated immunosuppressive effects. However, the involvement of MAPK independent pathways has not been clearly elucidated; nor has the crucial role played by LT in the early stages of infection. Determining whether LT exerts any pathological effects before being enriched to an MEK inhibitory level is an important next step in the furtherance of this field. METHODOLOGY/PRINCIPAL FINDINGS: Using a cell culture model, we determined that low doses of LT inhibited phagocytosis of macrophages, without influencing MAPK pathways. Consistent low doses of LT significantly suppressed bacterial clearance and enhanced the mortality of mice with bacteremia, without suppressing the MEK1 of splenic and peripheral blood mononuclear cells. CONCLUSION/SIGNIFICANCE: These results suggest that LT suppresses the phagocytes in a dose range lower than that required to suppress MEK1 in the early stages of infection

Capsule synthesis by Bacillus anthracis is required for dissemination in murine inhalation anthrax

Bacillus anthracis, the agent of anthrax, produces a poly-D-glutamic acid capsule that has been implicated in virulence. Many strains missing pXO2 (96 kb), which harbors the capsule biosynthetic operon capBCAD, but carrying pXO1 (182 kb) that harbors the anthrax toxin genes, are attenuated in animal models. Also, noncapsulated strains are readily phagocytosed by macrophage cell lines, whereas capsulated strains are resistant to phagocytosis. We show that a strain carrying both virulence plasmids but deleted specifically for capBCAD is highly attenuated in a mouse model for inhalation anthrax. The parent strain and capsule mutant initiated germination in the lungs, but the capsule mutant did not disseminate to the spleen. A mutant harboring capBCAD but deleted for the cap regulators acpA and acpB was also significantly attenuated, in agreement with the capsule-negative phenotype during in vitro growth. Surprisingly, an acpB mutant, but not an acpA mutant, displayed an elevated LD(50) and reduced ability to disseminate, indicating that acpA and acpB are not true functional homologs and that acpB may play a larger role in virulence than originally suspected

Near Eradication of Clinically Relevant Concentrations of Human Tumor Cells by Interferon-Activated Monocytes In Vitro

We have previously reported that low concentrations of interferon (IFN)-activated monocytes exert near-eradicative cytocidal activity against low concentrations of several human tumor cells in vitro. In the present study, we examined 7 human tumor cell lines and 3 diploid lines in the presence or absence of 10 ng/mL IFNα2a and monocytes. The results confirmed strong cytocidal activity against 4 of 7 tumor lines but none against 3 diploid lines. To model larger in vivo tumors, we increased the target cell concentration and determined the concentration of IFNα2a and monocytes, required for cell death. We found that increasing the tumor cell concentration from 10- to 100-fold (105 cells/well) required an increase in the concentration of IFNs by over 100-fold and monocytes by 10-fold. High concentrations of monocytes could sometimes kill tumor or diploid cells in the absence of IFN. We may conclude that killing of high concentrations of tumor or diploid cells required high concentrations of monocytes that could sometimes kill in the absence of IFN. Thus, high concentrations of tumor cells required high concentrations of IFN and monocytes to cause near eradication of tumor cells. These findings may have clinical implications

A Novel Multiscale Methodology forSimulating Tunnel Ventilation Flows duringFire

This paper applies a novel and fast modelling approach to simulate tunnel ventilation flows during fires. The complexity and high cost of full CFD models and the inaccuracies of simplistic zone or analytical models are avoided by efficiently combining mono-dimensional (1D) and CFD (3D) modelling techniques. A simple 1D network approach is used to model tunnel regions where the flow is fully developed (far field), and a detailed CFD representation is used where flow conditions require 3D resolution (near field). This multi-scale method has previously been applied to simulate tunnel ventilation systems including jet fans, vertical shafts and portals (Colella et al., Build Environ 44(12): 2357-2367, 2009) and it is applied here to include the effect of fire. Both direct and indirect coupling strategies are investigated and compared for steady state conditions. The methodology has been applied to a modern tunnel of 7 m diameter and 1.2 km in length. Different fire scenarios ranging from 10 MW to 100 MW are investigated with a variable number of operating jet fans. Comparison of cold flow cases with fire cases provides a quantification of the fire throttling effect, which is seen to be large and to reduce the flow by more than 30% for a 100 MW fire. Emphasis has been given to the discussion of the different coupling procedures and the control of the numerical error. Compared to the full CFD solution, the maximum flow field error can be reduced to less than few percents, but providing a reduction of two orders of magnitude in computational time. The much lower computational cost is of great engineering value, especially for parametric and sensitivity studies required in the design or assessment of ventilation and fire safety system