Single-cell RNA transcriptomics reveals differences in the immune status of alcoholic and hepatitis B virus-related liver cirrhosis

Abstract

BackgroundAlcoholic and hepatitis B virus (HBV)-related liver cirrhosis has placed a tremendous burden on the healthcare system with limited treatment options. This study explored the differences in the immune status of alcoholic and HBV-related liver cirrhosis.MethodsA total of 15 human liver samples from the Third Xiangya Hospital of Central South University, including five healthy controls (HC group), five alcoholic cirrhosis patients (ALC group), and five HBV-related cirrhosis patients (HBV group) were used. Of these, eight samples, including 3 HC group, 2 ALC group and 3 HBV group, were randomly collected to do single-cell RNA sequencing (scRNA-seq). The degree of steatosis was assessed by H&E staining and the presence of intrahepatic immune cells was evaluated by immunochemistry (IHC).ResultsThe immune status of alcoholic and HBV-related liver cirrhosis differed significantly. ScRNA-seq analysis identified a higher ratio of intrahepatic monocyte/macrophages and an obvious decreased ratio of T cells and B cells in the ALC group than in the HBV group. IHC staining of intrahepatic monocyte/macrophages, T and B cell exhibited similar results with scRNA-seq analysis. CD5L+ Kupffer cells, a cell type involved in lipid metabolism, were the major monocyte/macrophage subset in ALC liver tissue. H&E staining indicated that the level of steatosis was more severe in the ALC than in the HBV group. Ligand/receptor analysis showed that the T cell exhaustion observed in the ALC liver may be related to the expression of Galectin-9 on Kupffer cells. Fewer B cells were also found in the ALC group and most had higher lipid metabolism, reduced ribosomal activity, and a dysregulated mitochondrial oxidative phosphorylation system. Moreover, scRNA-seq showed a significantly lower ratio of plasma B cells, indicating that the humoral immune response in the ALC liver was similarly dysfunctional. Ligand/receptor analysis also discovered that Galectin-9 expressed on Kupffer cells may inhibit humoral immunity.ConclusionPatients with ALC have different immune characteristics than those with HBV-induced cirrhosis, including an increased ratio of intrahepatic monocyte/macrophages and a dysfunctional adaptive immune response in the liver. Galectin-9 could serve as a potential therapeutic target for ALC treatment.

Single-cell Technologies Provide Novel Insights into Liver Physiology and Pathology

“…The proportion of T and B cells was significantly lower, which might be related to the expression of Galectin-9 in Kupffer cells. 53 Therefore, the immune profile of ALC patients differs from that of HBV cirrhotic patients, and Galectin-9 might be a therapeutic target for treating ALC.…”

“…49 Studies using single-cell technology have provided valuable information on the characteristics of single cells and their gene-expression profile of normal liver and liver diseases from multiple perspectives (Table 2). [50][51][52][53][54][55][56][57][58][59][60][61][62] As Wang et al 63 provided a comprehensive single-cell immune profile of HBV-infected patients, revealing that increased Treg and Tex cells are associated with the degree of liver damage, and that immune cell interactions networks may promote persistent HBV infection and liver histopathology. Thus, HBV infection is a dynamic process, and a deeper understanding of cellular interactions and the changes of immune microenvironment will help develop more effective therapeutic targets and biomarkers for immunotherapy in chronic hepatitis B (CHB) patients, thereby slowing down the occurrence of end-stage liver disease (Fig.…”