Salmonella Enteritidis antitoxin DinJ inhibits NLRP3-dependent canonical inflammasome activation in macrophages

炎症体 肠炎沙门氏菌 生物 NLRC4型 先天免疫系统 分泌物 半胱氨酸蛋白酶1 免疫系统 微生物学 细胞生物学 沙门氏菌 炎症 免疫学 遗传学 生物化学 细菌
作者
Dan Gu,Ang Li,Xirui Zang,Tingting Huang,Yaxin Guo,Xinan Jiao,Zhiming Pan
出处
期刊:Infection and Immunity [American Society for Microbiology]
标识
DOI:10.1128/iai.00505-23
摘要

ABSTRACT The inflammasome is a pivotal component of the innate immune system, acting as a multiprotein complex that plays an essential role in detecting and responding to microbial infections. Salmonella Enteritidis have evolved multiple mechanisms to regulate inflammasome activation and evade host immune system clearance. Through screening S . Enteritidis C50336Δ fliC transposon mutant library, we found that the insertion mutant of dinJ increased inflammasome activation. In this study, we demonstrated the genetic connection between the antitoxin DinJ and the toxin YafQ in S . Enteritidis, confirming their co-transcription. The deletion mutant Δ fliC Δ dinJ increased cell death and IL-1β secretion in J774A.1 cells. Western blotting analysis further showed elevated cleaved Caspase-1 product (p10 subunits) and IL-1β secretion in cells infected with Δ fliC Δ dinJ compared to cells infected with Δ fliC . DinJ was found to inhibit canonical inflammasome activation using primary bone marrow-derived macrophages (BMDMs) from Casp -/- C57BL/6 mice. Furthermore, DinJ specifically inhibited NLRP3 inflammasome activation, as demonstrated in BMDMs from Nlrp3 -/- and Nlrc4 -/- mice. Fluorescence resonance energy transfer (FRET) experiments confirmed the translocation of DinJ into host cells during infection. Finally, we revealed that DinJ could inhibit the secretion of IL-1β and IL-18 in vivo , contributing to S . Enteritidis evading host immune clearance. In summary, our findings provide insights into the role of DinJ in modulating the inflammasome response during S . Enteritidis infection, highlighting its impact on inhibiting inflammasome activation and immune evasion.
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