Objective: To explore the roles of the expression of the co-stimulatory molecule, B7-2, and the co-inhibitory molecule, PD-L1, on peripheral blood mononuclear cells in the mechanism of immunotolerance in chronic hepatitis B virus infection. Methods: Thirty HBV infected patients in the immunoreactive phase and 20 patients in the immunotolerant phase were enrolled in the study, while 20 healthy volunteers were used as controls. RT- PCR and real-time PCR methods were used to detect the expression levels of B7-2 and PD-L1 mRNA in peripheral blood mononuclear cells in chronic HBV infected patients. Results: The B7-2 expression in immunoreactive and immunotolerant patients was significantly lower than that in the controls (P all < 0.01 ); B7-2 expression in immunoreactive patients was significantly lower than in immunotolerant patients (P < 0.01). PD-L1 expression in immunoreactive patients and immunotolerant patients was significantly higher than that in normal controls (P all < 0.01). The PD-L1/B7-2 ratios in immunoreactive and immunotolerant patients were significantly higher than that of the healthy controls (P all < 0.01); the PD-L1/B7-2 ratio was significantly higher in the immunoreactive patients than in the immunotolerant patients (P < 0.01). Conclusion: In chronic HBV infection, changes in the expression of co-stimulatory and co-inhibitory molecules imply a protective adjustment against the patient's immune response that may result in increased immunotolerance and persistent HBV infection.