Reassessing the Evolutionary Importance of Inflammasomes

Inflammasomes monitor the cytosol for microbial contamination or perturbation, and are thus predicted to provide potent defense against infection. However, the compendium of data from murine infection models suggests that inflammasomes merely delay the course of disease, allowing the host time to mount an adaptive response. Interpretations of such results are confounded by inflammasome evasion strategies of vertebrate-adapted pathogens. Conversely, environmental opportunistic pathogens have not evolved in the context of inflammasomes, and are therefore less likely to evade them. Indeed, opportunistic pathogens do not normally cause disease in wild type animals. Accordantly, the extreme virulence of two opportunistic bacterial pathogens, Burkholderia thailandensis and Chromobacterium violaceum, is fully counteracted by inflammasomes in murine models. This leads us to propose a new hypothesis: perhaps animals maintain inflammasomes over evolutionary time not to defend against vertebrate-adapted pathogens, but instead to counteract infection by a plethora of undiscovered opportunistic pathogens residing in the environment

Programmed cell death as a defence against infection

Eukaryotic cells can die from physical trauma, resulting in necrosis. Alternately, they can die via programmed cell death upon stimulation of specific signalling pathways. Here we discuss the utility of four cell death pathways in innate immune defence against bacterial and viral infection: apoptosis, necroptosis, pyroptosis and NETosis. We describe the interactions that interweave different programmed cell death pathways, which create complex signalling networks that cross-guard each other in the evolutionary arms race with pathogens. Finally, we describe how the resulting cell corpses — apoptotic bodies, pore-induced intracellular traps (PITs) and neutrophil extracellular traps (NETs) — promote clearance of infection

Pyroptosis triggers pore-induced intracellular traps (PITs) that capture bacteria and lead to their clearance by efferocytosis

Inflammasomes activate caspase-1 in response to cytosolic contamination or perturbation. This inflammatory caspase triggers the opening of the GSDMD pore in the plasma membrane, resulting in lytic cell death called pyroptosis. We had previously assumed that pyroptosis releases intracellular bacteria to the extracellular space. Here, we find that viable bacteria instead remain trapped within the cellular debris of pyroptotic macrophages. This trapping appears to be an inevitable consequence of how osmotic lysis ruptures the plasma membrane, and may also apply to necroptosis and some forms of nonprogrammed necrosis. Although membrane tears release soluble cytosolic contents, they are small enough to retain organelles and bacteria. We call this structure the pore-induced intracellular trap (PIT), which is conceptually parallel to the neutrophil extracellular trap (NET). The PIT coordinates innate immune responses via complement and scavenger receptors to drive recruitment of and efferocytosis by neutrophils. Ultimately, this secondary phagocyte kills the bacteria. Hence, caspase-1–driven pore-induced cell death triggers a multifaceted defense against intracellular bacteria facilitated by trapping the pathogen within the cellular debris. Bona fide intracellular bacterial pathogens, such as Salmonella , must prevent or delay pyroptosis to avoid being trapped in the PIT and subsequently killed by neutrophils

Salmonella and Caspase-1: A complex Interplay of Detection and Evasion

Salmonellae are intracellular pathogens that replicate within epithelial cells and macrophages, and are a significant public health threat in both developed and developing countries. The innate immune system detects microbes through pattern recognition receptors, which are compartmentalized on the subcellular level to detect either extracellular (e.g., TLRs) or cytosolic (e.g., NLRs) perturbations. Salmonella infection is detected by the NLRC4 and NLRP3 inflammasomes, which activate Caspase-1, resulting in reduced bacterial burdens during infection. NLRC4 responds to the SPI1 type III secretion system via detection of inadvertently translocated flagellin and rod protein. The signals for NLRP3 detection during Salmonella infection remain undefined. Salmonella have evolved evasion strategies to attenuate Caspase-1 responses. We review recent findings describing the interplay between detection and evasion of S. typhimurium infection by the inflammasome. We discuss how the interplay between detection and evasion affects Caspase-1 effector functions mediated by IL-1β secretion, IL-18 secretion, and pyroptosis

IL-1β, IL-18, and eicosanoids promote neutrophil recruitment to pore-induced intracellular traps following pyroptosis

Inflammasomes activate caspase-1, initiating a lytic form of programmed cell death termed pyroptosis, which is an important innate immune defense mechanism against intracellular infections. We recently demonstrated in a mouse infection model of pyroptosis that instead of releasing bacteria into the extracellular space, bacteria remain trapped within the pyroptotic cell corpse, termed the pore-induced intracellular trap (PIT). This trapping mediates efferocytosis of the PIT and associated bacteria by neutrophils; bacteria are subsequently killed via neutrophil ROS. Using this pyroptosis model, we now show that the pro-inflammatory cytokines IL-1β and IL-18 and inflammatory lipid mediators termed eicosanoids are required for effective clearance of bacteria downstream of pyroptosis. We further show that IL-1β, IL-18, and eicosanoids affect this in part by mediating neutrophil recruitment to the PIT. This is in addition to our prior findings that complement is also important to attract neutrophils. Thus, the PIT initiates a robust and coordinated innate immune response involving multiple mediators that attract neutrophils to efferocytose the PIT and its entrapped bacteria

Pyroptotic cell death defends against intracellular pathogens

Inflammatory caspases play a central role in innate immunity by responding to cytosolic signals and initiating a twofold response. First, caspase-1 induces the activation and secretion of the two prominent pro-inflammatory cytokines, interleukin-1β (IL-1β) and IL-18. Second, either caspase-1 or caspase-11 can trigger a form of lytic, programmed cell death called pyroptosis. Pyroptosis operates to remove the replication niche of intracellular pathogens, making them susceptible to phagocytosis and killing by a secondary phagocyte. However, aberrant, systemic activation of pyroptosis in vivo may contribute to sepsis. Emphasizing the efficiency of inflammasome detection of microbial infections, many pathogens have evolved to avoid or subvert pyroptosis. This review focuses on molecular and morphological characteristics of pyroptosis and the individual inflammasomes and their contribution to defense against infection in mice and humans

Detection of cytosolic bacteria by inflammatory caspases

The sanctity of the cytosolic compartment is rigorously maintained by a number of innate immune mechanisms. Inflammasomes detect signatures of microbial infection and trigger caspase-1 or caspase-11 activation, culminating in cytokine secretion and obliteration of the replicative niche via pyroptosis. Recent studies have examined inflammatory caspase responses to cytosolic bacteria, including Burkholderia, Shigella, Listeria, Francisella, and Mycobacterium species. For example, caspase-11 responds to LPS introduced into the cytosol after Gram-negative bacteria escape the vacuole. Not surprisingly, bacteria antagonize these responses; for example, Shigella delivers OspC3 to inhibit caspase-4. These findings underscore bacterial coevolution with the innate immune system, which has resulted in few, but highly specialized cytosolic pathogens

The development of high field magnets utilizing Bi-2212 wind & react insert coils

Wind & react Bi-2212 inserts have been manufactured and tested inside a wide-bore NbTi-Nb3Sn magnet providing a background field up to 20T at 4.2K. A pair of six-layer concentric coils both achieved critical currents of 350 A (JE = 200 A/mm2) in a 20T background field. A thicker 14-layer insert made from 119m of round wire had a critical quench current IQ of 287A (JE = 162 A/mm2) at the same field and contributed to a combined central field of 22.5 T. This is a record for a fully superconducting magnet at 4.2 K. The 14-layer coil, equipped with an external protective shunt, was used for an extensive series of quench measurements and endured >150 quenches without damage. Minimum quench energies were found to be in the range of 200-500mJ in background fields of 15-20T when the coil carried 70-95% of its critical quench current

Salmonella and Caspase-1: A complex Interplay of Detection and Evasion

Salmonellae are intracellular pathogens that replicate within epithelial cells and macrophages, and are a significant public health threat in both developed and developing countries. The innate immune system detects microbes through pattern recognition receptors, which are compartmentalized on the subcellular level to detect either extracellular (e.g., TLRs) or cytosolic (e.g., NLRs) perturbations. Salmonella infection is detected by the NLRC4 and NLRP3 inflammasomes, which activate Caspase-1, resulting in reduced bacterial burdens during infection. NLRC4 responds to the SPI1 type III secretion system via detection of inadvertently translocated flagellin and rod protein. The signals for NLRP3 detection during Salmonella infection remain undefined. Salmonella have evolved evasion strategies to attenuate Caspase-1 responses. We review recent findings describing the interplay between detection and evasion of S. typhimurium infection by the inflammasome. We discuss how the interplay between detection and evasion affects Caspase-1 effector functions mediated by IL-1β secretion, IL-18 secretion, and pyroptosis

Autophagy May Allow a Cell to Forbear Pyroptosis When Confronted With Cytosol-Invasive Bacteria

Inflammatory caspases detect cytosol-invasive Gram-negative bacteria by monitoring for the presence of LPS in the cytosol. This should provide defense against the cytosol-invasive Burkholderia and Shigella species by lysing the infected cell via pyroptosis. However, recent evidence has shown caspase-11 and gasdermin D activation can result in two different outcomes: pyroptosis and autophagy. Burkholderia cepacia complex has the ability invade the cytosol but is unable to inhibit caspase-11 and gasdermin D. Yet instead of activating pyroptosis during infection with these bacteria, the autophagy pathway is stimulated through caspases and gasdermin D. In contrast, Burkholderia thailandensis can invade the cytosol where caspasae-11 and gasdermin D is activated but the result is pyroptosis of the infected cell. In this review we propose a hypothetical model to explain why autophagy would be the solution to kill one type of Burkholderia species, but another Burkholderia species is killed by pyroptosis. For pathogens with high virulence, pyroptosis is the only solution to kill bacteria. This explains why some pathogens, such as Shigella have evolved methods to inhibit caspase-11 and gasdermin D as well as autophagy. We also discuss similar regulatory steps that affect caspase-1 that may permit the cell to forbear undergoing pyroptosis after caspase-1 activates in response to bacteria with partially effective virulence factors