Anthrax lethal toxin and Salmonella elicit the common cell death pathway of caspase-1-dependent pyroptosis via distinct mechanisms

Molecular and Cellular Biology Program, Departments of Microbiology and Laboratory Medicine, University of Washington, Seattle, WA 98195, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 04/2008; 105(11):4312-7. DOI: 10.1073/pnas.0707370105
Source: PubMed


Caspase-1 cleaves the inactive IL-1beta and IL-18 precursors into active inflammatory cytokines. In Salmonella-infected macrophages, caspase-1 also mediates a pathway of proinflammatory programmed cell death termed "pyroptosis." We demonstrate active caspase-1 diffusely distributed in the cytoplasm and localized in discrete foci within macrophages responding to either Salmonella infection or intoxication by Bacillus anthracis lethal toxin (LT). Both stimuli triggered caspase-1-dependent lysis in macrophages and dendritic cells. Activation of caspase-1 by LT required binding, uptake, and endosome acidification to mediate translocation of lethal factor (LF) into the host cell cytosol. Catalytically active LF cleaved cytosolic substrates and activated caspase-1 by a mechanism involving proteasome activity and potassium efflux. LT activation of caspase-1 is known to require the inflammasome adapter Nalp1. In contrast, Salmonella infection activated caspase-1 through an independent pathway requiring the inflammasome adapter Ipaf. These distinct mechanisms of caspase-1 activation converged on a common pathway of caspase-1-dependent cell death featuring DNA cleavage, cytokine activation, and, ultimately, cell lysis resulting from the formation of membrane pores between 1.1 and 2.4 nm in diameter and pathological ion fluxes that can be blocked by glycine. These findings demonstrate that distinct activation pathways elicit the conserved cell death effector mechanism of caspase-1-mediated pyroptosis and support the notion that this pathway of proinflammatory programmed cell death is broadly relevant to cell death and inflammation invoked by diverse stimuli.

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    • "Extracellular ATP activates the ATP-gated ion channel P2X7 and triggers rapid K þ efflux (Franchi et al. 2007; Petrilli et al. 2007); Nigericin creates a K þ pore in the cell membrane (Mariathasan et al. 2006); the influenza M2 protein triggers export of H þ ions from the Golgi complex into the cytosol (Ichinohe et al. 2010); and high concentrations of extracellular Ca 2þ , increase cytosolic Ca 2þ , and cAMP (Lee et al. 2012; Murakami et al. 2012; Rossol et al. 2012). However, it is important to note that ion fluxes also activate other inflammasomes , such as NLRP1b (Fink et al. 2008; Wickliffe et al. 2008; Newman et al. 2009; Ali et al. 2011) and NLRC4 (Arlehamn et al. 2010). This suggests that ion fluxes might only modulate the threshold of caspase-1 activation but not serve as specific signals that trigger NLRP3 inflammasome activation (Lamkanfi and Dixit 2012). "
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    ABSTRACT: Inflammasomes are large cytosolic multiprotein complexes that assemble in response to detection of infection- or stress-associated stimuli and lead to the activation of caspase-1-mediated inflammatory responses, including cleavage and unconventional secretion of the leaderless proinflammatory cytokines IL-1β and IL-18, and initiation of an inflammatory form of cell death referred to as pyroptosis. Inflammasome activation can be induced by a wide variety of microbial pathogens and generally mediates host defense through activation of rapid inflammatory responses and restriction of pathogen replication. In addition to its role in defense against pathogens, recent studies have suggested that the inflammasome is also a critical regulator of the commensal microbiota in the intestine. Finally, inflammasomes have been widely implicated in the development and progression of various chronic diseases, such as gout, atherosclerosis, and metabolic syndrome. In this perspective, we discuss the role of inflammasomes in infectious and noninfectious inflammation and highlight areas of interest for future studies of inflammasomes in host defense and chronic disease.
    Cold Spring Harbor perspectives in biology 10/2014; 6(12). DOI:10.1101/cshperspect.a016287 · 8.68 Impact Factor
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    • "Manipulation of cell-death modality has been successfully used by other intracellular pathogens such as Chlamydia, Legionella pneumophila, Listeria monocytogenes, Shigella flexineri, and Salmonella enterica subsp. enterica serovar Typhimurium [28-30]. It has been demonstrated that host-cell apoptosis confers protection to the host, once the uptake of apoptotic bodies derived from macrophages by dendritic cells allows an effective activation of the immune response [31]. "
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    ABSTRACT: Background Phospholipases C (PLCs) are virulence factors found in several bacteria. In Mycobacterium tuberculosis (Mtb) they exhibit cytotoxic effects on macrophages, but the mechanisms involved in PLC-induced cell death are not fully understood. It has been reported that induction of cell necrosis by virulent Mtb is coordinated by subversion of PGE2, an essential factor in cell membrane protection. Results Using two Mtb clinical isolates carrying genetic variations in PLC genes, we show that the isolate 97-1505, which bears plcA and plcB genes, is more resistant to alveolar macrophage microbicidal activity than the isolate 97-1200, which has all PLC genes deleted. The isolate 97-1505 also induced higher rates of alveolar macrophage necrosis, and likewise inhibited COX-2 expression and PGE2 production. To address the direct effect of mycobacterial PLC on cell necrosis and PGE2 inhibition, both isolates were treated with PLC inhibitors prior to macrophage infection. Interestingly, inhibition of PLCs affected the ability of the isolate 97-1505 to induce necrosis, leading to cell death rates similar to those induced by the isolate 97-1200. Finally, PGE2 production by Mtb 97-1505-infected macrophages was restored to levels similar to those produced by 97-1200-infected cells. Conclusions Mycobacterium tuberculosis bearing PLCs genes induces alveolar macrophage necrosis, which is associated to subversion of PGE2 production.
    BMC Microbiology 05/2014; 14(1):128. DOI:10.1186/1471-2180-14-128 · 2.73 Impact Factor
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    • "Briefly, MVs were sized by comparison to calibrations of the flow cytometer using beads ranging from <0.1–1 micron from Spherotech Inc, IL using the manufacture's protocol. The flow cytometry data had previously been validated by transmission electron microscopy (TEM) as described in our previous work [26]. Pelleted MVs were then subjected directly to western blotting, caspase-1 enzymatic assay or added to fresh lymphocytes. "
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    ABSTRACT: Immune dysregulation during sepsis is poorly understood, however, lymphocyte apoptosis has been shown to correlate with poor outcomes in septic patients. The inflammasome, a molecular complex which includes caspase-1, is essential to the innate immune response to infection and also important in sepsis induced apoptosis. Our group has recently demonstrated that endotoxin-stimulated monocytes release microvesicles (MVs) containing caspase-1 that are capable of inducing apoptosis. We sought to determine if MVs containing caspase-1 are being released into the blood during human sepsis and induce apoptosis.. Single-center cohort study. 50 critically ill patients were screened within 24 hours of admission to the intensive care unit and classified as either a septic or a critically ill control. Circulatory MVs were isolated and analyzed for the presence of caspase-1 and the ability to induce lymphocyte apoptosis. Patients remaining in the ICU for 48 hours had repeated measurement of caspase-1 activity on ICU day 3. Septic patients had higher microvesicular caspase-1 activity 0.05 (0.04, 0.07) AFU versus 0.0 AFU (0, 0.02) (p<0.001) on day 1 and this persisted on day 3, 0.12 (0.1, 0.2) versus 0.02 (0, 0.1) (p<0.001). MVs isolated from septic patients on day 1 were able to induce apoptosis in healthy donor lymphocytes compared with critically ill control patients (17.8±9.2% versus 4.3±2.6% apoptotic cells, p<0.001) and depletion of MVs greatly diminished this apoptotic signal. Inhibition of caspase-1 or the disruption of MV integrity abolished the ability to induce apoptosis. These findings suggest that microvesicular caspase-1 is important in the host response to sepsis, at least in part, via its ability to induce lymphocyte apoptosis. The ability of microvesicles to induce apoptosis requires active caspase-1 and intact microvesicles.
    PLoS ONE 03/2014; 9(3):e90968. DOI:10.1371/journal.pone.0090968 · 3.23 Impact Factor
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