Toll-like receptors (TLRs) are a class of proteins that play key roles in innate immunity through recognition of microbial components. TLR3 is expressed abundantly in dendritic cells, and is responsible for recognizing viral pathogens and inducing interferon beta (IFN-beta) production. Although TLR3 has been reported to be involved in several diseases caused by viral infections, its role in hepatitis B virus (HBV)-induced hepatitis is still largely unknown. We found that expression of TLR3 and IFN-beta was decreased significantly in monocyte-derived dendritic cells (MoDCs) from patients with chronic hepatitis B (CHB, n=40) or acute-on-chronic hepatitis B liver failure (ACHBLF, n=60), compared with normal controls (n=20). We observed a further decrease in TLR3 and IFN-beta in ACHBLF compared to CHB patients. Compared with surviving patients, TLR3 and IFN-beta expression was significantly lower in non-surviving ACHBLF patients, which strongly indicated a correlation between TLR3 signaling impairment in MoDCs and disease severity in ACHBLF patients. Further linear correlation analysis demonstrated significant correlations between expression of TLR3 signaling components (TLR3 and IFN-beta) and disease severity markers (prothrombin activity and total bilirubin) for individual ACHBLF patients. To the best of our knowledge, this is the first study to show that MoDC impairment is correlated with severe liver damage in ACHBLF patients, which suggests the potential of TLR3/IFN-beta expression in MoDCs as a diagnostic marker.