Enteropathogenic Escherichia coli Uses NleA to Inhibit NLRP3 Inflammasome Activation

PLoS Pathogens
Hilo YenToru Tobe

Abstract

Enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC) are related strains capable of inducing severe gastrointestinal disease. For optimal infection, these pathogens actively modulate cellular functions through the deployment of effector proteins in a type three secretion system (T3SS)-dependent manner. In response to enteric pathogen invasion, the Nod-like receptor pyrin domain containing (NLRP) inflammasome has been increasingly recognized as an important cytoplasmic sensor against microbial infection by activating caspase-1 and releasing IL-1β. EPEC and EHEC are known to elicit inflammasome activation in macrophages and epithelial cells; however, whether the pathogens actively counteract such innate immune responses is unknown. Using a series of compound effector-gene deletion strains of EPEC, we screened and identified NleA, which could subdue host IL-1β secretion. It was found that the reduction is not because of blocked NF-κB activity; instead, the reduction results from inhibited caspase-1 activation by NleA. Immunostaining of human macrophage-like cells following infection revealed limited formation of inflammasome foci with constituents of total caspase-1, ASC and NLRP3 in the presence of NleA. Pulldo...Continue Reading

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Methods Mentioned

BETA
ELISA
nuclear translocation
confocal microscopy
immunoprecipitation
ubiquitination
co-immunoprecipitation
pulldown
pull down
de-ubiquitination
transfection

Software Mentioned

Adobe
FLUOVIEW Viewer
Adobe Photoshop
Photoshop

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