Objective：To explore the role of negative regulators of toll-like receptor（TLRs） signal pathways in immunological patho-genesis of severe hepatitis B virus infection（CHB）. Methods：mRNA expressions of myeloid-differentiation-88-short（MyD88s），interleukin-1R-associated-kinase-M（IRAK-M），single-immunoglobulin-interleukin-1R-related-molecule（SIGIRR），zinc finger protein A20 and TLR4 in peripheral blood mononuclear cells（PBMCs） in 20 cases of chronic CHB，17 cases of acute-on-chronic liver failure at early stage（ACLF-E），9 cases of acute-on-chronic liver failure at late stage（ACLF-L） and 18 healthy controls were detected by real time fluorescence quantitative PCR. Serum levels of tumor necrosis factor-α（TNF-α） and interleukin-10（IL-10） were detected by ELISA assay. Results：Compared with those in healthy controls，mRNA expression levels of MyD88s，IRAK-M and A-20 as well as serum levels of TNF-α and IL-10 were unregulated with the progression of diseases in CHB patients，ACLF-E patients and ACLF-L patients（P<0.05）（P<0.01）. There was no significant difference in serum levels of IL-10 between CHB and ACLF-E（P >0.05）. mRNA expressions of TLR4 were higher in CHB patients，ACLF-E patients and ACLF-L patients than in healthy controls. mRNA expressions of TLR4 were lower in ACLF-L patients than in ACLF-E patients（P<0.05）. Conclusions：Negative regulators of TLRs signaling pathway participate in the pathogenesis of chronic hepatitis and liver failure and upregulation of negative regulators may lead to immunosuppressive in patients with liver failure.