Pseudorabies virus DNA binding to porcine NLRC3 unleashes the activation of the porcine NLRP3 inflammasome

Minjie Li; Xiangyu Huang; Hongyan Yin; Lvye Chai; Haiwei Wang; Xin Li

doi:10.1016/j.vetmic.2025.110604

Pseudorabies virus DNA binding to porcine NLRC3 unleashes the activation of the porcine NLRP3 inflammasome

Vet Microbiol. 2025 Jun 13:307:110604. doi: 10.1016/j.vetmic.2025.110604. Online ahead of print.

Authors

Minjie Li¹, Xiangyu Huang¹, Hongyan Yin¹, Lvye Chai¹, Haiwei Wang², Xin Li³

Affiliations

¹ National Key Laboratory of Veterinary Public Health and Safety, China Agricultural University, Beijing 100193, China; Key Laboratory of Animal Epidemiology of the Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, China Agricultural University, Beijing, China.
² State Key Laboratory of Animal Disease Control, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China.
³ National Key Laboratory of Veterinary Public Health and Safety, China Agricultural University, Beijing 100193, China; Key Laboratory of Animal Epidemiology of the Ministry of Agriculture and Rural Affairs, College of Veterinary Medicine, China Agricultural University, Beijing, China. Electronic address: xinli2021@cau.edu.cn.

PMID: 40526995
DOI: 10.1016/j.vetmic.2025.110604

Abstract

Viral infection activates multiple inflammatory pathways, with the NLRP3 inflammasome playing a pivotal role in host defense. However, negative regulation of the NLRP3 inflammasome is essential for maintaining host homeostasis. Here, we report that double-stranded DNA (dsDNA) from pseudorabies virus (PRV) induces NLRP3 inflammasome activation and pyroptosis through gasdermin D (GSDMD) cleavage and IL-1β secretion. Importantly, the inhibitory NLR porcine NLRC3 (pNLRC3) interacts with porcine NLRP3 (pNLRP3) and attenuates GSDMD cleavage and IL-1β release. Upon PRV infection, overexpression of pNLRC3 enhances GSDMD cleavage and lactate dehydrogenase release, whereas knockdown of pNLRC3 reduces pyroptosis. Mechanistically, pNLRC3 binds PRV dsDNA and unleashes its inhibitory effect on pNLRP3, functioning as a checkpoint to regulate inflammasome activation. Furthermore, pNLRC3 contributes to PRV restriction by controlling viral replication and limiting infection. In summary, our findings reveal a dual role of pNLRC3, acting both as a negative regulator of the pNLRP3 inflammasome and as a viral sensor that regulates pyroptosis-mediated viral clearance. These insights provide a deeper understanding of virus-host interactions and innate immune regulation.

Keywords: Inflammasome; Porcine NLRC3; Porcine NLRP3; Pseudorabies virus; Pyroptosis.