PGLYRP2 drives hepatocyte-intrinsic innate immunity by trapping and clearing hepatitis B virus
Ying Li, … , Yuanfei Yao, Ming Shi
Ying Li, … , Yuanfei Yao, Ming Shi
Published February 13, 2025
Citation Information: J Clin Invest. 2025;135(8):e188083. https://doi.org/10.1172/JCI188083.
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Research Article Hepatology Virology

PGLYRP2 drives hepatocyte-intrinsic innate immunity by trapping and clearing hepatitis B virus

Abstract

Spontaneous clearance of hepatitis B virus (HBV) is frequent in adults (95%) but rare in infants (5%), emphasizing the critical role of age-related hepatic immunocompetence. However, the underlying mechanisms of hepatocyte-specific immunosurveillance and age-dependent HBV clearance remain unclear. Here, we identified PGLYRP2 as a hepatocyte-specific pattern recognition receptor with age-dependent expression, and demonstrated that phase separation of PGLYRP2 was a critical driver of spontaneous HBV clearance in hepatocytes. Mechanistically, PGLYRP2 recognized and potentially eliminated covalently closed circular DNA via phase separation, coordinated by its intrinsically disordered region and HBV DNA-binding domain (PGLYRP2IDR/209–377) in the nucleus. Additionally, PGLYRP2 suppressed HBV capsid assembly by directly interacting with the viral capsid, mediated by its PGRP domain. This interaction promoted the nucleocytoplasmic translocation of PGLYRP2 and subsequent secretion of the PGLYRP2/HBV capsid complex, thereby bolstering the hepatic antiviral response. Pathogenic variants or deletions in PGLYRP2 impaired its ability to inhibit HBV replication, highlighting its essential role in hepatocyte-intrinsic immunity. These findings suggest that targeting the PGLYRP2-mediated host-virus interaction may offer a potential therapeutic strategy for the development of anti-HBV treatments, representing a promising avenue for achieving a functional cure for HBV infection.

Authors

Ying Li, Huihui Ma, Yongjian Zhang, Tinghui He, Binyang Li, Haoran Ren, Jia Feng, Jie Sheng, Kai Li, Yu Qian, Yunfeng Wang, Haoran Zhao, Jie He, Huicheng Li, Hongjin Wu, Yuanfei Yao, Ming Shi

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Figure 4

HBV capsid interacts with PGLYRP2 and induces nucleocytoplasmic translocation in HBV-infected cells.

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HBV capsid interacts with PGLYRP2 and induces nucleocytoplasmic transloc...
(A) Localization of PGLYRP2 (red) and HBc (green) in HBV or HBc expression construct–transfected Huh7/PGLYRP2 cells was analyzed by immunofluorescence staining. Scale bars: 20 μm. (B) HBV– or HBV+ human liver tissue lysates were prepared for co-IP using anti-PGLYRP2 and blotted using the indicated antibodies. (C) Schematic representation of WT and mutants of HBc (C61G and Y132A). (D) PGLYRP2 expression construct was cotransfected with Con vector, HBc WT, HBc-C61G, or HBc-Y132A expression construct into HEK293 cells. After 48 hours, cell lysates were harvested for co-IP using anti-FLAG antibody and blotted using the indicated antibodies. The high-order complex in the whole-cell lysates (WCLs) was separated by native PAGE and subjected to immunoblot. (E) Nucleocytoplasmic translocation of PGLYRP2 (red) in WT HBc, HBc-C61G, or HBc-Y132A mutant (green) expression construct–transfected Huh7/PGLYRP2 cells was analyzed by immunofluorescence staining. Scale bars: 20 μm. (F) Left: Representative confocal images showing PLA+ and PLA– signal in HBV+ liver tissue or HBV– liver tissue, respectively. Top right: A PLA signal corresponds to PGLYRP2/HBV capsid proximity, whereas the absence of HBV capsid resulted in the lack of a PLA fluorescent signal. Bottom right: Quantification of percentage of PLA+ cells from the total number of detected cells. Dots indicate biological replicates (n = 5 for HBV– tissues and n = 9 for HBV+ tissues; independent experiments). Scale bars: 20 μm. Data are represented as mean ± SD. Two-tailed Student’s t test was used for statistical analysis. **P < 0.001. (G) Our proposed model of PGLYRP2NLS masking. The AlphaFold3-predicted structure model of PGLYRP2 and the 3D structure of PGLYRP2/HBV capsid (Protein Data Bank: 6VZP) complex were visualized by PyMOL software (https://pymol.org).