Pyroptosis induced by TLR9 ligand, ODN1826, requires ROS production and caspase-11 activation in Raw264.7 cells
Issued Date
2025-09-01
Resource Type
ISSN
0125877X
eISSN
22288694
Scopus ID
2-s2.0-105020784666
Pubmed ID
37302095
Journal Title
Asian Pacific Journal of Allergy and Immunology
Volume
43
Issue
3
Start Page
745
End Page
752
Rights Holder(s)
SCOPUS
Bibliographic Citation
Asian Pacific Journal of Allergy and Immunology Vol.43 No.3 (2025) , 745-752
Suggested Citation
Pudla M., Saengfak R., Luangjindarat C., Utaisincharoen P. Pyroptosis induced by TLR9 ligand, ODN1826, requires ROS production and caspase-11 activation in Raw264.7 cells. Asian Pacific Journal of Allergy and Immunology Vol.43 No.3 (2025) , 745-752. 752. doi:10.12932/ap-041222-1511 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/113005
Title
Pyroptosis induced by TLR9 ligand, ODN1826, requires ROS production and caspase-11 activation in Raw264.7 cells
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Abstract
Background: Toll-like receptor 9 (TLR9), located in the endosomal compartment, is known to play a role in inflammation by recognizing oligonucleotides that contain CpG motive (CpG-ODN). Signaling by TLR9 leads to the production of proinflammatory cytokines and can trigger cell death. Objective: This study aims to investigate the molecular mechanism of pyroptosis induced by ODN1826 in the mouse macrophage cell line (Raw264.7). Methods: The protein expression and the amount of lactate dehydrogenase (LDH) of ODN1826-treated cells were determined by immunoblotting and LDH assay, respectively. In addition, the level of cytokine production was observed by ELISA assay and the ROS production was determined by flow cytometry. Results: Our results showed that ODN1826 induced pyroptosis as judged by LDH releases. Furthermore, caspase-11 and gasdermin D activation, which are the key molecules in pyroptosis, were also observed in ODN1826-activated cells. Moreover, we also demonstrated that Reactive Oxygen Species (ROS) production by ODN1826 is essential for caspase-11 activation and gasdermin D release, which leads to pyroptosis. Conclusion: ODN1826 induces pyroptosis in Raw264.7 cells via caspase-11 and GSDMD activation. Moreover, the production of ROS by this ligand plays an essential role in the regulation of caspase-11 and GSDMD activation, which then controls pyroptosis in TLR9 activation.