Methods: 25 patients were recruited in the trial including 18 with ALT≤2ULN and 7 with ALT>2ULN. PBMCs obtained from 50ml of heparinized peripheral blood through density gradient centrifuge and adherence method were proliferated under the induction by GM-CSF and IL-4, and sensitized with the stock of hepatitis B vaccine containing 30µg HBsAg on day 5 and with hepatitis B vaccine commercially available containing 20µg HBsAg on day 6. anti-HBV-DC vaccine was harvested on day 7 and injected, half hypodermically and half intravenously, to the patient once every two weeks for 12 practices applications totally. Thymosin-α1 1.6mg was injected hypodermically twice a week. Quantitative HBVM(TRFIA) and HBVDNA and hepatic functions were evaluated at week 0, 4, 12, and 24. 

Results: Mean of HBsAg, ATL and TBIL decreased gradually along the time from week 4, 12 to 24, with significant difference compared with the values prior to treatment. At week 4, 12 and 24, HBsAg negative conversion rate were 8.00%(2/25), 12.00(3/25) and 20.00%(5/25) respectively, HBVDNA negative conversion rate were 63.64%(7/11), 72.73%(8/11) and 72.73%(8/11), ALT normalization rate were 48.00%(12/25), 64.00%(16/25) and 80.00%(20/25), and HBsAg negative conversion rate was 11.11%(2/18), 16.67%(3/18) and 22.22%(4/18) in patients with ALT≤2ULN. But HBsAg negative conversion occurred only in one patient (14.29%, 1/7) those with ALT>2ULN at week 24. The rate of adverse effect was 2.67% observed in reinfusion of anti-HBV-DC vaccine. 

Conclusions: anti-HBV-DC vaccine in combination with thymosin-α1 can be considered as a safe approach with high efficacy for HBeAg negative CHB patients, which can effectively inhibit the viral replication, decrease rapidly and eliminate the HBsAg and HBVDNA from body.