Aspergillus fumigatus Stimulates the NLRP3 Inflammasome through a Pathway Requiring ROS Production and the Syk Tyrosine Kinase

Invasive aspergillosis (IA) is a life-threatening disease that occurs in immunodepressed patients when infected with Aspergillus fumigatus. This fungus is the second most-common causative agent of fungal disease after Candida albicans. Nevertheless, much remains to be learned about the mechanisms by which A. fulmigatus activates the innate immune system. We investigated the inflammatory response to conidia and hyphae of A. fumigatus and specifically, their capacity to trigger activation of an inflammasome. Our results show that in contrast to conidia, hyphal fragments induce NLRP3 inflammasome assembly, caspase-1 activation and IL-1β release from a human monocyte cell line. The ability of Aspergillus hyphae to activate the NLRP3 inflammasome in the monocytes requires K+ efflux and ROS production. In addition, our data show that NLRP3 inflammasome activation as well as pro-IL-1β expression relies on the Syk tyrosine kinase, which is downstream from the pathogen recognition receptor Dectin-1, reinforcing the importance of Dectin-1 in the innate immune response against fungal infection. Furthermore, we show that treatment of monocytes with corticosteroids inhibits transcription of the gene encoding IL-1β. Thus, our data demonstrate that the innate immune response against A. fumigatus infection involves a two step activation process, with a first signal promoting expression and synthesis of pro-IL-1β; and a second signal, involving Syk-induced activation of the NLRP3 inflammasome and caspase-1, allowing processing and secretion of the mature cytokine

Targeting of a Chlamydial Protease Impedes Intracellular Bacterial Growth

Chlamydiae are obligate intracellular bacteria that propagate in a cytosolic vacuole. Recent work has shown that growth of Chlamydia induces the fragmentation of the Golgi apparatus (GA) into ministacks, which facilitates the acquisition of host lipids into the growing inclusion. GA fragmentation results from infection-associated cleavage of the integral GA protein, golgin-84. Golgin-84-cleavage, GA fragmentation and growth of Chlamydia trachomatis can be blocked by the peptide inhibitor WEHD-fmk. Here we identify the bacterial protease chlamydial protease-like activity factor (CPAF) as the factor mediating cleavage of golgin-84 and as the target of WEHD-fmk-inhibition. WEHD-fmk blocked cleavage of golgin-84 as well as cleavage of known CPAF targets during infection with C. trachomatis and C. pneumoniae. The same effect was seen when active CPAF was expressed in non-infected cells and in a cell-free system. Ectopic expression of active CPAF in non-infected cells was sufficient for GA fragmentation. GA fragmentation required the small GTPases Rab6 and Rab11 downstream of CPAF-activity. These results define CPAF as the first protein that is essential for replication of Chlamydia. We suggest that this role makes CPAF a potential anti-infective therapeutic target

TLR2/MyD88/NF-κB Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection

Human respiratory syncytial virus (RSV) constitute highly pathogenic virus that cause severe respiratory diseases in newborn, children, elderly and immuno-compromised individuals. Airway inflammation is a critical regulator of disease outcome in RSV infected hosts. Although “controlled” inflammation is required for virus clearance, aberrant and exaggerated inflammation during RSV infection results in development of inflammatory diseases like pneumonia and bronchiolitis. Interleukin-1β (IL-1β) plays an important role in inflammation by orchestrating the pro-inflammatory response. IL-1β is synthesized as an immature pro-IL-1β form. It is cleaved by activated caspase-1 to yield mature IL-1β that is secreted extracellularly. Activation of caspase-1 is mediated by a multi-protein complex known as the inflammasome. Although RSV infection results in IL-1β release, the mechanism is unknown. Here in, we have characterized the mechanism of IL-1β secretion following RSV infection. Our study revealed that NLRP3/ASC inflammasome activation is crucial for IL-1β production during RSV infection. Further studies illustrated that prior to inflammasome formation; the “first signal” constitutes activation of toll-like receptor-2 (TLR2)/MyD88/NF-κB pathway. TLR2/MyD88/NF-κB signaling is required for pro-IL-1β and NLRP3 gene expression during RSV infection. Following expression of these genes, two “second signals” are essential for triggering inflammasome activation. Intracellular reactive oxygen species (ROS) and potassium (K+) efflux due to stimulation of ATP-sensitive ion channel promote inflammasome activation following RSV infection. Thus, our studies have underscored the requirement of TLR2/MyD88/NF-κB pathway (first signal) and ROS/potassium efflux (second signal) for NLRP3/ASC inflammasome formation, leading to caspase-1 activation and subsequent IL-1β release during RSV infection

STATIC AND DYNAMIC STABILITY OF GRAVI-INERTIAL SENSORS WITH CAPACITIVE DIFFERENTIAL SYSTEM OF SENSITIVITY ADJUSTMENT

The design of a gravi-inertial sensor with an elastically suspended sensing mass exhibiting a maximum sensitivity and minimum noise level is being studied. It is conceived that such a sensor contains a torsion mass-spring system, a capacitive pick-off circuit to detect motions of the sensing mass, and a capacitive system to reduce torsion stiffness. Both capacitive systems are combined into a single differential capacitance electrostatic system. The torsion stiffness is reduced by applying an electric field. Problems resulting from the electrostatic asymmetry of the differential system are studied analytically and numerically. The quasi-static and dynamic modes of the free movement of the sensing mass, in the absence of energy loss, are considered. The angular intervals of stability of the sensing mass movement in the electrostatic field, depending on the differential system asymmetry parameter and a frequency of free oscillations «proof mass», are calculated

СТАТИЧЕСКАЯ И ДИНАМИЧЕСКАЯ СТАБИЛЬНОСТЬ ГРАВИИНЕРЦИАЛЬНОГО ДАТЧИКА С ЕМКОСТНОЙ ДИФФЕРЕНЦИАЛЬНОЙ СИСТЕМОЙ УПРАВЛЕНИЯ ЧУВСТВИТЕЛЬНОСТЬЮ

The design of a gravi-inertial sensor with an elastically suspended sensing mass exhibiting a maximum sensitivity and minimum noise level is being studied. It is conceived that such a sensor contains a torsion mass-spring system, a capacitive pick-off circuit to detect motions of the sensing mass, and a capacitive system to reduce torsion stiffness. Both capacitive systems are combined into a single differential capacitance electrostatic system. The torsion stiffness is reduced by applying an electric field. Problems resulting from the electrostatic asymmetry of the differential system are studied analytically and numerically. The quasi-static and dynamic modes of the free movement of the sensing mass, in the absence of energy loss, are considered. The angular intervals of stability of the sensing mass movement in the electrostatic field, depending on the differential system asymmetry parameter and a frequency of free oscillations «proof mass», are calculated.Изучается конструкция гравиинерциального датчика с упруго подвешенной массой, обладающего максимальной чувствительностью и минимальным уровнем шума. Датчик содержит упругий торсион, электростатическую емкостную систему измерения движения подвижной массы и электростатическую емкостную систему уменьшения крутильной жесткости торсиона. Обе емкостные системы объединены в единую дифференциальную емкостную электростатическую систему. Аналитически и численно изучаются проблемы, возникающие в результате асимметрии этой системы. Исследуются квазистатический и динамический режимы свободного движения подвижной массы при отсутствии потерь энергии. Рассчитываются угловые интервалы устойчивости движения подвижной массы в электростатическом поле в зависимости от параметра асимметрии дифференциальной системы и от частоты свободных колебаний.