Abstract
Inflammasomes, which are intracellular sensors of endogenous or exogenous danger signals, activate caspase-1, resulting in interleukin (IL)-1β maturation. Although most studies on inflammasomes have been performed in human and/or mouse-derived macrophages, porcine inflammasome activation has not been elucidated even though pigs are considered one of the best animal models for translational and preclinical investigations. In this study, we optimized detection of porcine IL-1β secretion, which is the most well established indicator of inflammasome activation, and compared inflammasome activation between miniature and domestic pigs as well as between porcine and murine macrophages. In our results, anti-sera against murine IL-1β had higher affinity to porcine IL-1β than anti-sera against human IL-1β, even though the amino acid sequence of porcine IL-1β was more similar to that of human IL-1β. In addition, there was no significant difference in inflammasome activation between miniature and domestic pigs. Furthermore, well established inflammasome triggers (ATP, nigericin, and crystals) in humans and mice had similar effects on porcine NLRP3 inflammasome activation. We further elucidated the upstream signaling pathway of porcine inflammasome activation using pharmacological inhibitors. Similar to the mechanisms of inflammasome activation in humans and mice, potassium efflux and reactive oxygen species generation were confirmed as key pathways in porcine inflammasome activation. Thus, inflammasome activation in pigs is not different from that in humans or mice.
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Acknowledgments
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2012R1A1A1001645), by a grant from the Next-Generation BioGreen 21 Program (No. PJ009069), Rural Development Administration, Republic of Korea
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All animal experiments were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and approved by the Institutional Animal Care and Use Committee of Kangwon National University (ACUCC; approval no. KIACUC-12-0131).
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Jeeyoung Kim and Huijeong Ahn contributed equally to this work.
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Kim, J., Ahn, H., Woo, HM. et al. Characterization of porcine NLRP3 inflammasome activation and its upstream mechanism. Vet Res Commun 38, 193–200 (2014). https://doi.org/10.1007/s11259-014-9602-5
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DOI: https://doi.org/10.1007/s11259-014-9602-5