AIM:To investigate whether hepatitis B virus (HBV) could induce a hepatitis B virus core antigen (HBcAg)specific cytotoxic T lymphocyte (CTL) response in vitro by dendritic cells (DCs) transduced with lentiviral vector-encoding ubiquitinated hepatitis B virus core antigen (LV-Ub-HBcAg).METHODS:Recombinant LV-Ub-HBcAg were transfected into highly susceptible 293 T cells to obtain high virus titres.Bone marrow-derived DCs isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleukin (IL)-4.LV-Ub-HBcAg,lentiviral vector-encoding hepatitis B virus core antigen (LV-HBcAg),lentiviral vector (LV) or lipopolysaccharide were added to induce DC maturation,and the DC phenotypes were analyzed by flow cytometry.The level of IL-12 in the supernatant was detected by enzyme-linked immunosorbent assay (ELISA).T lymphocytes were proliferated using Cell Counting Kit-8.DCs were cultured and induced to mature using different LVs,and co-cultured with allogeneic T cells to detect the secretion levels of IL-2,IL-4,IL-10 and interferon-γ in the supernatants of T cells by ELISA.Intracellular cytokines of proliferative T cells were analyzed by flow cytometry,and specific CTL activity was measured by a lactate dehydrogenase release assay.RESULTS:LV-Ub-HBcAg-induced DCs secreted more IL-12 and upregulated the expression of CD80,CD86 and major histocompatibility class Ⅱ.DCs sensitised by different LVs effectively promoted cytokine secretion;the levels of IL-2 and interferon-γ induced by LV-UbHBcAg were higher than those induced by LV-HBcAg.Compared with LV-HBcAg-transduced DCs,LV-UbHBcAg-transduced DCs more efficiently stimulated the proliferation of T lymphocytes and generated HBcAgspecific cytotoxic T lymphocytes.CONCLUSION:LV-Ub-HBcAg effectively induced DC maturation.The mature DCs efficiently induced T cell polarisation to Th1 and generated HBcAg-specific CTLs.
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