Enteropathogenic Escherichia coli Uses NleA to Inhibit NLRP3 Inflammasome Activation

PLoS Pathogens

Hilo Yen¹  Toru Tobe¹  Nakaba Sugimoto² 

[1] Department of Biomedical Informatics, Graduate School of Medicine, Osaka University, Osaka, Japan;Department of Microbiology and Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan

关键词: Inflammasomes;    Secretion;    Immunoprecipitation;    Cell differentiation;    Transcription factors;    Immunoblotting;    Pathogens;    Antibodies;   

DOI  :  10.1371/journal.ppat.1005121

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

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. Pulldown of PMA-induced differentiated THP-1 lysate with purified MBP-NleA reveals that NLRP3 is a target of NleA. The interaction was verified by an immunoprecipitation assay and direct interaction assay in which purified MBP-NleA and GST-NLRP3 were used. We further showed that the effector interacts with regions of NLRP3 containing the PYD and LRR domains. Additionally, NleA was found to associate with non-ubiquitinated and ubiquitinated NLRP3 and to interrupt de-ubiquitination of NLRP3, which is a required process for inflammasome activation. Cumulatively, our findings provide the first example of EPEC-mediated suppression of inflammasome activity in which NieA plays a novel role in controlling the host immune response through targeting of NLRP3.

【 授权许可】

CC BY