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PD-L1 as a Potential Inducer for NF-κB Pathway Activation in M1-Type Macrophages During Sepsis: An Integrated
Mu Yuan1, Tong Jin1, Qing Zhao1
1Department of Stem Cell & Regenerative Medicine, State Key Laboratory of Trauma and Chemical Poisoning, Daping Hospital, Army Medical University, Chongqing, 400042, People's Republic of China.
Background:
M1-type macrophages are a crucial defense line against pathogens during sepsis, and their polarization depends on NF-κB pathway activation. However, the specific molecular regulatory mechanisms of this process are not fully elucidated. Programmed death-ligand 1 (PD-L1), an important immune checkpoint molecule, is often anchored on the cell membrane exerting immunosuppressive effects, but its role in M1-type macrophages remains poorly understood.
Methods:
In this study, we preliminarily investigated the potential regulatory effect of PD-L1 on the NF-κB pathway in M1 macrophages by integrating GEO sepsis patient sequencing datasets, RNA-seq data from PD-L1-overexpressing M1 macrophages, immunoprecipitation-mass spectrometry (IP-MS), molecular docking, and co-immunoprecipitation (Co-IP). The Receiver Operating Characteristic (ROC) curve of PD-L1 was used as a prognostic indicator for sepsis. PD-L1 knockout mice were utilized to validate the in vivo regulatory effects of PD-L1.
Results:
Analysis of GEO data and peripheral blood mononuclear cells (PBMCs) from sepsis patients revealed significantly elevated PD-L1 expression with a central position among all upregulated genes. Furthermore, this elevated PD-L1 expression was validated in peripheral monocytes from sepsis patients compared to healthy controls. Single-gene GSEA analysis based on GEO data indicated a close association between PD-L1 and NF-κB pathway activation in sepsis patients. RNA-seq analysis of PD-L1-overexpressing M1 macrophages confirmed that PD-L1 significantly activates the NF-κB pathway. IP-MS screening and Co-IP validation identified an interaction between PD-L1 and KLF6, a nuclear factor closely related to the NF-κB pathway. Finally, liver transcriptome data from septic mice confirmed that PD-L1 can also activate the NF-κB pathway in vivo, and ROC curve analysis demonstrated PD-L1's potential as a prognostic indicator for sepsis.
Conclusion:
Based on these results, this study preliminarily reveals a potential activating role of PD-L1 on the NF-κB pathway in M1 macrophages.
Insights
Programmed death-ligand 1 (PD-L1) activates the NF-κB pathway in M1 macrophages, crucial for sepsis defense. This study identifies PD-L1 as a potential prognostic indicator for sepsis, highlighting its role in immune regulation.
Area of Science:
- Immunology
- Molecular Biology
- Sepsis Pathophysiology
Background:
- M1 macrophages are key in sepsis defense, relying on NF-κB pathway activation for polarization.
- The precise molecular regulators of M1 macrophage polarization remain unclear.
- Programmed death-ligand 1 (PD-L1), an immune checkpoint molecule, has an incompletely understood role in M1 macrophages.
Purpose of the Study:
- To investigate the regulatory effect of PD-L1 on the NF-κB pathway in M1 macrophages.
- To explore PD-L1 as a prognostic biomarker for sepsis.
Main Methods:
- Integrated analysis of GEO sepsis datasets and RNA-seq data.
- Immunoprecipitation-mass spectrometry (IP-MS) and co-immunoprecipitation (Co-IP).
- Receiver Operating Characteristic (ROC) curve analysis and PD-L1 knockout mouse models.
Main Results:
- Elevated PD-L1 expression in sepsis patients and its association with NF-κB pathway activation.
- PD-L1 overexpression activates the NF-κB pathway in M1 macrophages.
- Identified an interaction between PD-L1 and KLF6, and confirmed PD-L1's in vivo role in activating the NF-κB pathway in sepsis.
Conclusions:
- PD-L1 preliminarily shows an activating role in the NF-κB pathway within M1 macrophages.
- PD-L1 demonstrates potential as a prognostic indicator for sepsis.
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