The success of phagosome degradation relies on the ability of phagocytes to regulate the maturation of phagosomes. However, its underlying molecular mechanisms remain poorly understood. Here, we identify the proton-activated chloride (PAC) channel as a key negative regulator of phagosome maturation. PAC deletion enhanced phagosomal acidification and protease activities, leading to augmented bacterial killing in large peritoneal macrophages (LPMs) upon Escherichia coli infection in mice. Surprisingly, phagosome degradation also stimulated STING-IRF3-IFN responses and inflammasome activation in LPMs, both of which are enhanced upon PAC deletion. The increased inflammasome activation induced the release of cleaved gasdermin D, which localized to the surface of bacteria in the peritoneum and further contributed to their killing. Finally, enhanced bacterial clearance by PAC-deficient LPMs reduced proinflammatory immune cell infiltration and peritoneal inflammation, resulting in improved survival in mice. Our study thus provides new insights into the molecular mechanism of phagosome maturation and the dynamics of host defense response following phagosome-mediated bacterial degradation in peritoneal macrophages.
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3 November 2025
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August 22 2025
Proton-activated chloride channel governs phagosome-mediated antibacterial immunity in peritoneal macrophages
In Special Collection:
Macrophages and Dendritic Cells in Health and Disease
Henry Yi Cheng
,
Henry Yi Cheng
(Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Project administration, Resources, Software, Supervision, Validation, Visualization, Writing - original draft, Writing - review & editing)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
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Jiachen Chu
,
Jiachen Chu
(Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Project administration, Resources, Supervision, Validation, Visualization)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
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Nathachit Limjunyawong
,
Nathachit Limjunyawong
(Methodology)
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
3Center of Research Excellence in Allergy and Immunology, Research Department, Faculty of Medicine Siriraj Hospital,
Mahidol University
, Bangkok, Thailand
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Jianan Chen
,
Jianan Chen
(Formal analysis, Project administration)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
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Yingzhi Ye
,
Yingzhi Ye
(Formal analysis)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
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Kevin Hong Chen
,
Kevin Hong Chen
(Data curation, Formal analysis, Investigation, Methodology, Resources, Validation, Writing - review & editing)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
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Nicholas Koylass
,
Nicholas Koylass
(Data curation)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
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Shuying Sun
,
Shuying Sun
(Funding acquisition)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
4Brain Science Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA
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Xinzhong Dong
,
Xinzhong Dong
(Methodology, Project administration, Writing - review & editing)
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
5Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA
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Zhaozhu Qiu
(Conceptualization, Funding acquisition, Project administration, Resources, Supervision, Writing - original draft, Writing - review & editing)
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
6Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Correspondence to Zhaozhu Qiu: [email protected]
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Henry Yi Cheng
https://orcid.org/0000-0003-4412-4235
Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Project administration, Resources, Software, Supervision, Validation, Visualization, Writing - original draft, Writing - review & editing
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
Jiachen Chu
https://orcid.org/0000-0001-8206-1146
Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Project administration, Resources, Supervision, Validation, Visualization
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
Nathachit Limjunyawong
https://orcid.org/0000-0002-9540-6171
Methodology
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
3Center of Research Excellence in Allergy and Immunology, Research Department, Faculty of Medicine Siriraj Hospital,
Mahidol University
, Bangkok, Thailand
Jianan Chen
https://orcid.org/0009-0003-7782-0672
Formal analysis, Project administration
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
Yingzhi Ye
https://orcid.org/0000-0002-7380-6529
Formal analysis
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
Kevin Hong Chen
https://orcid.org/0000-0002-5421-1538
Data curation, Formal analysis, Investigation, Methodology, Resources, Validation, Writing - review & editing
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
Nicholas Koylass
https://orcid.org/0000-0003-3078-1867
Data curation
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
Shuying Sun
https://orcid.org/0000-0002-8449-7370
Funding acquisition
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
4Brain Science Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Xinzhong Dong
https://orcid.org/0000-0002-9750-7718
Methodology, Project administration, Writing - review & editing
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
5Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Zhaozhu Qiu
https://orcid.org/0000-0002-9122-6077
Conceptualization, Funding acquisition, Project administration, Resources, Supervision, Writing - original draft, Writing - review & editing
1Department of Physiology, Pharmacology and Therapeutics,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
2Solomon H. Snyder Department of Neuroscience,
Johns Hopkins University School of Medicine
, Baltimore, MD, USA
6Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA
Correspondence to Zhaozhu Qiu: [email protected]
Disclosures: The authors declare no competing interests exist.
Received:
February 11 2025
Revision Received:
May 05 2025
Accepted:
June 24 2025
Online ISSN: 1540-9538
Print ISSN: 0022-1007
Funding
Funder(s):
National Institutes of Health
- Award Id(s): R35GM124824,R01NS118014,RF1NS134549
Funder(s):
McKnight Foundation
Funder(s):
Klingenstein-Simon Scholar award
Funder(s):
Sloan Foundation
Funder(s):
Karen Toffler Charitable Trust
© 2025 Cheng et al.
2025
Cheng et al.
This article is distributed under the terms as described at https://rupress.org/pages/terms102024/.
J Exp Med (2025) 222 (11): e20250312.
Article history
Received:
February 11 2025
Revision Received:
May 05 2025
Accepted:
June 24 2025
Citation
Henry Yi Cheng, Jiachen Chu, Nathachit Limjunyawong, Jianan Chen, Yingzhi Ye, Kevin Hong Chen, Nicholas Koylass, Shuying Sun, Xinzhong Dong, Zhaozhu Qiu; Proton-activated chloride channel governs phagosome-mediated antibacterial immunity in peritoneal macrophages. J Exp Med 3 November 2025; 222 (11): e20250312. doi: https://doi.org/10.1084/jem.20250312
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