Background: Hepatocellular carcinoma (HCC) is one of the most common tumors in China, which frequently occurs in the patients with chronic hepatitis B virus infection.Although early diagnosis and surgery may improve survival, HCC is rarely cured and with high risk of recurrence.Adoptive T cell transfer (ACT) is an emerging treatment for various cancers including HCC.However, the complex procedures to manipulate autologous tumor-infiltrating lymphocytes or tumor antigen-specific T cell clones largely limit the application of ACT in clinical practices.Here we present a novel strategy named "Smart T" to prepare activated T lymphocytes in vitro, which contain HCC-specific cytotoxic T lymphocytes (CTLs) and with a promising outcome in a preliminary trial with HCC patients.Methods:PBMCs were isolated from 100mL peripheral blood of HCC patient.The adherent monocytes were continued to be cultured in AIM-V inedium with GM-CSF and IL-4to differentiate into immature dendritic cells (DCs).On day 5, the immature DCs were incubated with multiple HCC associated T cell epitope peptides pool along with diverse TLR ligands to differentiate into mature DCs.Meanwhile, the non-adherent PBMCs were maintained in AIM-V medium with low level of CD3 monoclonal antibody and IL-2.From day 7, the non-adherent PBMCs were co-culture with mature DCs for another 7 days in the presence of cytokines.Patients received infusions of the resulting T cells every 2-3 months with 5-10x107cells/kg body weight.Results: The resulting cells are mainly CD3+ T cells (96.65%±2.99%), with 78.98%±5.88% of CD8+ T cells.A large proportion of CD8+ T cells expressed NKG2D (76.6%±7.44%) and Granzyme B (40.4%±12.9%), indicating great potential of killing."Smart T"generated from HLA-A2+ patient exhibited greater cytotoxic activity to the HCC cell line HepG2 (HLA-A2+) than to HuH-7 cells (HLA-A2-).After 3 cycles of"Smart T" infusion, a significant increase was observed in the patients PBMCs for the frequency of CD3+ (P =0.025), CD8+ (P =0.023), CD8+NKG2D+ (P =0.043), CD8+CD107a+ (P =0.0082), especially effector T cells (P =0.0013) and central memory T cells (P =0.0038).In contrast, a significant decrease of regulatory T cells was observed (P =0.0015).Clinical measurements detected a better response (analyzed as CR, PR, SD and PD) in patients (N=19) who received multiple cycles (≧ 3) infusion of "Smart T", as compared to patients (N=19) who received less than 2 cycles infusion of "Smart T" (P =0.0259).No toxicity was observed.Conclusions: Our study, for the first time, demonstrates that "Smart T" strategy of ACT is a safe treatment which may improve the immunologic function and clinical responses of the HCC patients.