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Telbivudine (LdT) is an orally L-nucleoside with potent and specific antihepatitis B virus (HBV) activity. The higher rate of hepatitis B e antigen (HBeAg) seroconversion of LdT treatment than other anti-HBV agents suggests a potential immunomodulatory effect. The aim of the study was to investigate the changes of regulatory T cell (Treg)/interleukin (IL)-17-producing CD4+T helper (Th17) balance during LdT treatment and to discuss the relationship of Treg/Th17 balance with HBeAg change in HBeAg-positive chronic hepatitis B (CHB) patients receiving LdT antiviral treatment. Twenty-seven HBeAg-positive CHB patients received LdT for 24 weeks and the percentages of Tregs and cells (Th17 cells) in peripheral blood as well as the serum TGF-β1 and IL-17 levels in these patients were longitudinally analyzed. We found that the frequencies of Tregs and Th17 cells in peripheral blood as well as the serum TGF-β1 and IL-17 levels increased significantly in CHB patients compared with healthy controls. During the LdT treatment, the Tregs frequency and TGF-β1 level tended to decrease, and Th17 cells frequency and IL-17 level showed a reverse “V”-type change. The frequency of Tregs and the ratio of Treg/Th17 were significantly lower in the HBeAg loss group than those in the HBeAg no-loss group at the baseline. More important, the Tregs frequency and TGF-β1 level were both positively correlated with HBeAg level during the LdT treatment for 24 weeks. Our data suggest that the lower Tregs frequency and Treg/Th17 ratio at the baseline of LdT treatment, the more likely to get the HBeAg loss. HBeAg negative can be predicted using changes in Tregs frequency and TGF-β1 level during LdT treatment in CHB patients. Maybe we could provide the immunology marker for exploring the mechanism of the higher HBeAg seroconversion rate of LdT therapy.