Single-cell landscape of immune cells during the progression from HBV infection to HBV cirrhosis and HBV-associated hepatocellular carcinoma

Front Immunol. 2023 Dec 5:14:1320414. doi: 10.3389/fimmu.2023.1320414. eCollection 2023.

Abstract

Introduction: Immune cells play crucial roles in the development of chronic hepatitis B virus (HBV) infection, leading to cirrhosis and hepatocellular carcinoma (HCC). However, their functions at different disease stages are not fully understood.

Methods: In this study, we used single-cell RNA sequencing (scRNA-seq) to characterize the human liver immune microenvironment at different disease stages. We analyzed scRNA-seq data from 118,455 immune cells obtained from livers of six healthy individuals, four patients with HBV infection, five patients with HBV cirrhosis, and three patients with HBV-associated HCC.

Results: Our results showed an accumulation of scar-associated macrophages during disease progression, and we identified two relevant immune subsets, Macrophage-CD9/IL18 and macrophage-CD9/IFI6. Macrophage-CD9/IL18 expanded from HBV infection to cirrhosis, while macrophage-CD9/IFI6 expanded from cirrhosis to HCC. We verified the existence of Macrophage-CD9/IFI6 using multiplex immunofluorescence staining. We also found an increase in cytotoxic NK Cell-GNLY during progression from cirrhosis to HCC. Additionally, the proportion of CD4 T cell-TNFAIP3, CD8 T cell-TNF (effector CD8 T cells), and CD8 T cell-CD53 increased, while the proportion of Treg cells decreased from HBV infection to cirrhosis. The proportion of Treg and CD8 T cell-LAG3 (Exhausted CD8 T cell) enhanced, while the proportion of CD8 T cell-TNF (effector CD8 T cells) decreased from cirrhosis to HCC. Furthermore, GSEA enrichment analyses revealed that MAPK, ERBB, and P53 signaling pathways in myeloid cells were gradually inhibited from HBV infection to cirrhosis and HCC.

Discussion: Our study provides important insights into changes in the hepatic immune environment during the progression of HBV-related liver disease, which may help improve the management of HBV-infected liver diseases.

Keywords: HBV infection; cirrhosis; hepatitis B; hepatocellular carcinoma; scRNA-seq.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Hepatocellular*
  • Hepatitis B virus
  • Hepatitis B* / complications
  • Hepatitis B, Chronic*
  • Humans
  • Interleukin-18
  • Liver Cirrhosis / complications
  • Liver Neoplasms*
  • Tumor Microenvironment

Substances

  • Interleukin-18

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study was financially supported by Shenzhen Science and Technology Program (JCYJ20210324121802008), the National Natural Science Foundation of China (32270833), and the Natural Science Foundation of Fujian Province (2022J01014), the Hubei Technological Innovation Special Fund (CXPJJH122002-063), the Postdoctoral Foundation of Hei Long Jiang Province (LBH-Z20196), China Postdoctoral Science Foundation (2021MD703835); Wu JiePing Medical Foundation (320.6750.2021-23-22), the Open Fund of Key Laboratory of Hepatosplenic Surgery Ministery of Education Harbin China (GPKF202204), The Scientifc Foundation of the First Afliated Hospital of Harbin Medical University (Grant No.2023B12).