ht to be part of the very first line of CNS defense against bacterial infection as many Gram-positive and Gram-negative meningeal pathogens induce expression of these genes in hBMEC, van Sorge et al. unpublished data). Active impairment of neutrophil recruitment could therefore benefit survival and proliferation of B. anthracis, as both spores and vegetative bacteria are efficiently killed by human neutrophils. Our results clearly demonstrate that the suppression of CXCL1 and IL-8 expression is pXO1- and toxin-dependent, respectively. These data 1215493-56-3 complement observations in recent studies where systemic infection with the encapsulated strain impaired production of cytokines in a toxin-dependent manner and purified LT reduced IL-8 production by the destabilization 2067001 of IL-8 mRNA in HUVEC in vitro. We hypothesized that altered chemokine expression would result in impaired neutrophil recruitment upon active infection with B. anthracis Sterne. Using two independent in vivo assays, we demonstrated that neutrophil chemotaxis was indeed reduced to the site of infection with the Sterne strain as compared to infection with the DLF/EF mutant. Similar observations were recently published in a systemic infection model using encapsulated WT B. anthracis; host neutrophil recruitment in spleen and liver was significantly increased in the absence anthrax toxins compared to infection with the parent strain. In addition, purified LT has been shown to directly impair neutrophil motility. Toxin-mediated subversion of the innate immune system, specifically targeting neutrophils, may therefore contribute to unchecked bacterial replication and a more fulminent disease course. Establishment of an anthrax meningitis model is critical to better understand disease pathogenesis. The current rabbit and rhesus monkey models of inhalation anthrax both report signs of meningitis in a subgroup of animals, however, a mouse model would be preferable due to availability, lower costs and well-characterized genetic systems. We found that intravenous injection of immunocompetent outbred CD-1 mice with B. anthracis Sterne resulted in penetration of bacilli into the CNS. Microscopic analysis of brain sections confirmed the development of meningitis, showing inflammatory cell infiltration, hemorrhaging, thrombosis, edema and areas full of bacilli. While we did observe neutrophil infiltration in the brains of B. anthracis Sterne infected mice at the time of death, we speculate that an 
initial reduction or delay in host neutrophilic response may promote Toxins and Anthrax Meningitis acute unrestricted bacterial proliferation and further CNS dissemination ultimately responsible for the rapidly progressive deteriorating course associated with anthrax meningitis. These observations reflect autopsy findings in patients validating the utility of this newly developed mouse model 16507713 of hematogenous anthrax meningitis. Finally, development of anthrax meningitis requires expression of anthrax toxins as no signs of disease developed in mice infected with the DLF/EF mutant strain. 9 Toxins and Anthrax Meningitis Additional in vitro studies suggested that this could be due to a direct contribution of the toxins to penetration of brain endothelium; however, we cannot exclude the possibility that the lack of clinical symptoms observed during infection with the toxindeficient mutant may partially reflect a generalized reduction in virulence. In summary, our studies provide the first evidence that B. a
