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Objective: To study the effect of IFN-alpha2b on HBVDNA and inducement to CD25 of the patients with chronic hepatitis B. Methods: All of patients were divided into two groups and treated by IFN-alpha2b, routine medicine, respectively. The levels of HBV-DNA in PBMC and serum, and CD25 and CD25 mRNA, and anti-IFN-IgG were respectively detected by real-time PCR, biotin-streptavidin (BSA) and ELISA. Results: After treatment for 24 and 48 weeks, the total negative rates of HBV-DNA in PBMC, serum and HBeAg were 36.36%, 39.39%, 40.91% and 42.42%, 51.52%, 53.03%, respectively. There was significant difference between in the two groups (P < 0.05 ~ P <0.01). The level of CD25 either in silence or in inducement was low in chronic hepatitis B. Only was 4 (6.06%) cases with anti-IFN-IgG(+) and little discrepancy between in two groups (P > 0.05). Conclusions: IFN-alpha2b has better effect on HBV-DNA. CD25 can be induced by IFN-alpha2b so that the active T cells play key role against HBVDNA in hosts. Low level of anti-IFN-IgG can be produced during the treatment.