Objective:To study the effect of the Cytokine-induced killer cells cultured with engineered cells for costimulatory enhancement(ECCE-CIK) on inhibiting the replication of HBV produced by the human hepatoma cell lines HepG2.2.15. Methods:To generate ECCE-CIKs, ECCE were co-cultured with the peripheral blood mononuclear cells, IFN-γ, anti-CD3 antibody,and IL-2 were added into the culture system later. Then effector cells(ECCE-CIK) were co-cultured with target cells(HepG2.2.15) at effector to target (E:T) ratios of 1:1, 5:1, 20:1. After that the method of CFSE/PI was used to evaluate the death of target cells by flow cytometry. Then direct culturing system and indirect culturing system were established for the co-culturing of effector cells and target cells in vitro. At last the supernatants were collected from the two systems at 3, 24 and 48h respectively for detecting the amount of HBV DNA and HBsAg. Results:Accompanied by the increasement of E:T ratios and the duration of co-culturing the percentage of CFSE+ PI+ cells were increased progressively, while the levels of HBV DNA and HBsAg were decreased(P<0.05). Conclusion:These results suggestes that HBV replication in HepG2.2.15 can be suppressed by ECCE-CIK in both direct system and indirect system through cytolytic and noncytolytic mechanisms which may have a good clinical prospect.
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