Objective To observe the clinical efficacy of autologous dendritic cells (DCs) vaccine combined with cytokine-induced killer (CIK) cells for treatment of patients with biliary tract cancer. Methods Peripheral blood mononuclear cells (PBMCs) were collected from the patients with biliary tract cancer and were stimulated with multiple tumor-associated antigens (TAA) for amplification and culture. Mature DCs were harvested and made into vaccine for intradermal administration, and at the same time the patients were given autologous CIK cells via vein transfusion. The clinical efficacy, immune function, side effect, and overall survival of 85 patients with biliary tract cancer were observed before and after treatment. Results The clinical symptoms were improved significantly in the 85 patients after autologous DC-CIK therapy (P <0.05). The clinical efficacy was remarkable: with 2 cases (2.3%) having complete remission, 14 (16.5%) with partial remission, 54 (63.5%) with stable disease and 15 (17.6%) with progressive disease. The immune function was improved after combined treatment; CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, CD3⁺CD56⁺ and CD4⁺/CD8⁺ ratio were elevated in the lymhocyte subpopulation, but the regulatory T cells were reduced (P <0.05). At the end of the follow-up, 58 patients died, 2 patients were lost in follow-up, and 25 patients survived, with the median overall survival time being 16.5 months (95%CI: 12.1-20.9 months). Conclusion Autologous DC-CIK therapy is a safe and effective treatment for patients with biliary tract cancer, with less side effect, but the relevant conclusion still needs to be confirmed by larger sample randomized controlled studies.