CD100 is a positive regulator for immune responses. In HBV infection, CD100 can regulate T cells and macrophages activation for antiviral responses, raising the possibility that it might become a potential target for the treatment of HBV-related liver cirrhosis. However, the CD100 expression in HBV-related liver cirrhosis and its impact on hepatocytes have yet to be determined. In this study, The CD100 levels in the serum, platelets and immune cells from chronic hepatitis B (CHB) patients were tested. The effects of CD100 stimulation on primary hepatocytes were examined by mRNA-microarray analysis, while the influence of CD100 on extracellular matrix (ECM) deposition was assessed in a hepatocyte and hepatic stellate cell (HSCs) co-culture system. GO and PPI Enrichments were used to evaluate shared genes of CD100-regulated genes and changed genes during HBV-related liver cirrhosis. It is proved that the serum sCD100 and CD100 on platelets, which possess the potential to directly interact with hepatocytes, were down-regulated in CHB patients compared to healthy donors. CD100 expression on platelets was decreased in cirrhotic patients compared with non-cirrhotic patients. CD100 modulated gene expression in primary hepatocytes through plexin B1 and B2 signal transduction, which activated the ERK signaling pathway. The CD100 stimulation induced immune responses in hepatocytes, elevated the expression of TGF-β antagonists and HNF4A-related genes, and inhibited collagen I deposition in the hepatocytes and HSCs co-culture system. The effects may potentially serve as a drug target for the treatment of HBV-related liver cirrhosis.