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Updated: Jun 19, 2026
Ex Vivo Organotypic Corneal Model of Acute Epithelial Herpes Simplex Virus Type I Infection
Published on: November 3, 2012
Time-Course Transcriptomic Analysis Reveals PANoptosis-CCL2 Axis in Herpes Simplex Keratitis
Changyu Wu1, Junwen Ouyang1, Yaoyao Liu1
1Department of Ophthalmology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Purpose:
This study aimed to characterize the temporal dynamics of corneal immune responses and programmed cell death (PCD) following herpes simplex virus type 1 (HSV-1) infection, and investigate the role of PANoptosis in shaping the immune responses during herpes simplex keratitis (HSK).
Methods:
A murine HSK model was established through HSV-1 infection, and time-course RNA sequencing was performed on murine corneas at 0, 1, 3, 7, and 14 days post-infection. Phenotypic and mechanistic analyses were conducted using quantitative real-time PCR, western blotting, immunofluorescence staining, flow cytometry, and co-culture experiments. PCD pathways were assessed using antagonists targeting PANoptosis, an integrated form of cell death encompassing key features of apoptosis, pyroptosis, and necroptosis.
Results:
RNA sequencing revealed sequential immune response phases in the cornea following HSV-1 infection: homeostasis disruption and transcriptional activation (0-3 days), immune amplification (3-7 days), and tissue reorganization (7-14 days). The immune amplification phase was strongly associated with the activation of three distinct PCD pathways: apoptosis, pyroptosis, and necroptosis, culminating in PANoptosis. PANoptosis was confirmed as a central mechanism in HSK pathogenesis, with Z-DNA-binding protein 1 (ZBP1) serving as a key driver of PANoptosome formation. Furthermore, PANoptosis in human corneal epithelial cells (HCECs) was shown to promote the release of C-C motif chemokine ligand 2 (CCL2), enhancing macrophage major histocompatibility complex class II (MHC-II) and interferon β (IFN-β) expression and migration. The PANoptosis-CCL2/C-C motif chemokine receptor 2 (CCR2) axis was significantly upregulated in infected corneas and could be suppressed by a PANoptosis inhibitor or the CCL2 antagonist Bindarit.
Conclusions:
This study demonstrated the temporal coordination between immune activation and PANoptosis in HSK, highlighting PANoptosis as a crucial mechanism in amplifying antiviral immune responses. The ZBP1-PANoptosome-mediated PANoptosis-CCL2/CCR2 axis represents a potential therapeutic target for modulating the immune responses in HSK.
Insights
Herpes simplex keratitis involves sequential immune responses and programmed cell death (PCD), with PANoptosis amplifying antiviral immunity. Targeting the PANoptosis-CCL2/CCR2 axis offers a potential therapeutic strategy for HSK.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Herpes simplex virus type 1 (HSV-1) causes herpes simplex keratitis (HSK), a significant ocular infection.
- Understanding the interplay between immune responses and programmed cell death (PCD) is crucial for HSK pathogenesis.
- PANoptosis, an integrated form of PCD, has emerged as a key cellular process in various inflammatory conditions.
Purpose of the Study:
- To characterize the temporal dynamics of corneal immune responses and PCD after HSV-1 infection.
- To investigate the specific role of PANoptosis in modulating immune responses during HSK.
- To identify potential therapeutic targets within the identified pathways.
Main Methods:
- Established a murine model of HSK induced by HSV-1 infection.
- Performed time-course RNA sequencing on infected murine corneas.
- Utilized quantitative real-time PCR, western blotting, immunofluorescence, flow cytometry, and co-culture experiments for mechanistic analysis.
- Assessed PCD pathways using antagonists targeting PANoptosis.
Main Results:
- Identified sequential immune response phases: homeostasis disruption, immune amplification, and tissue reorganization.
- Demonstrated that the immune amplification phase involves apoptosis, pyroptosis, and necroptosis, culminating in PANoptosis.
- Confirmed Z-DNA-binding protein 1 (ZBP1) as a driver of PANoptosome formation in HSK.
- Showed that PANoptosis promotes C-C motif chemokine ligand 2 (CCL2) release, enhancing macrophage activation and migration via the CCL2/CCR2 axis.
- Found that the PANoptosis-CCL2/CCR2 axis is upregulated in infected corneas and can be suppressed by inhibitors.
Conclusions:
- HSK pathogenesis involves a temporal coordination between immune activation and PANoptosis.
- PANoptosis plays a critical role in amplifying antiviral immune responses during HSK.
- The ZBP1-PANoptosome-mediated PANoptosis-CCL2/CCR2 axis represents a promising therapeutic target for HSK treatment.
