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.
View: Text | PDF
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

×

Figure 1

Age-dependent expression of PGLYRP2 and its interactions with HBV promoter DNA.

Options: View larger image (or click on image) Download as PowerPoint
Age-dependent expression of PGLYRP2 and its interactions with HBV promot...
(A) Schematic overview of screening for HBV promoter–binding proteins. Diagram illustrating the experimental design for identifying proteins that bind to HBV promoter regions. (B) Identification of HBV promoter–binding host proteins. Proteins bound to HBV promoter DNA were isolated using a biotin-streptavidin affinity pull-down assay and visualized on a 12% SDS-PAGE gel with silver staining to confirm purity and presence. (C) ChIP assays were conducted to examine the interaction between PGLYRP2 and HBV promoter DNA. Anti-PGLYRP2 antibodies were used to pull down the relevant DNA-protein complexes from HBV+ and HBV– tissue lysates (Input). (D) Single-cell transcriptomic analysis by t-distributed stochastic neighbor embedding (t-SNE). Left: t-SNE plots illustrating the distribution of hepatocytes sampled at 4 distinct time points, displayed in 4 different colors. Right: A separate t-SNE plot highlights expression levels of PGLYRP2 across these cells. D, day; W, week. (E) Bar plots showing expression levels of PGLYRP2 in hepatocytes, as derived from the t-SNE analysis. The y axis represents log-normalized expression levels, emphasizing differences across time points. (F) Age-related expression of Pglyrp2 in mouse liver. Real-time PCR and Western blot analyses were used to measure PGLYRP2 levels across various age groups in mouse liver, illustrating an age-dependent expression pattern. (G) Dnmt3a expression analysis in mouse liver by age group. Real-time PCR was used to assess the expression of Dnmt3a across different age groups, revealing a decline in expression with age. Dots indicate biological replicates (n = 3 independent experiments). (H) Correlation between DNMT3A and PGLYRP2 expression. The relationship between Dnmt3a and Pglyrp2 mRNA levels was quantitatively analyzed in mouse liver tissues, highlighting a significant negative correlation. Data are represented as mean ± SD. Kruskal-Wallis with Dunn’s post hoc multiple-comparison test (E) and 1-way ANOVA with post hoc Bonferroni’s test (F and G) were used for statistical analysis. Pearson’s correlation coefficient was used in H. *P < 0.05; **P < 0.001.