Dendritic cell-based vaccination with lentiviral vectors encoding ubiquitinated hepatitis B core antigen enhances hepatitis B virus-specific immune responses <italic>in vivo</italic>

Abstract

The activity of hepatitis B virus (HBV)-specific cytotoxic T lymphocytes (CTLs) plays a predominant role in the clearance of HBV. Dendritic cells (DCs) are key antigen-presenting cells and play an important role in the initiation of immune responses. We previously verified that lentiviral vector encoding ubiquitinated hepatitis B core antigen (LV-Ub-HBcAg) effectively transduced DCs to induce maturation, and the mature DCs efficiently induced T cell polarization to Th1 and generated HBcAg-specific CTLs ex vivo. In this study, HBV-specific immune responses of LV-Ub-HBcAg in BALB/c mice (H-2K d ) were evaluated. It was shown that direct injection of LV-Ub-HBcAg increased the production of cytokines IL-2 and IFN-γ, elicited strong antibody responses, and remarkably generated a high percentage of IFN-γ + CD8 + T cells with HBV-specific CTL responses in BALB/c mice. In addition, direct injection of LV-Ub-HBcAg induced potent anti-HBV immune responses, similar to those elicited by in vitro-transduced DCs. In conclusion, the DC-based therapeutic vaccine LV-Ub-HBcAg elicited specific antibody immune responses and induced robust specific CTL activity in vivo.

An engineered novel lentivector specifically transducing dendritic cells and eliciting robust HBV-specific CTL response by upregulating autophagy in T cells

“…Lentivirus vectors (LVs) have been the focus of many studies because of the ability to transduce nondividing cells and permanently integrate into the genome of target cell with high efficiency [7,8]. We have previously confirmed that lentivirus-delivered and ubiquitin-fused Hepatitis B core antigen (LV-Ub-HBcAg) could promote the maturation of dendritic cells (DCs), trigger the preferential activation of HBV-specific CTL immune response and elicit therapeutic immunity in HBV transgenic mice [9][10][11]. Glycoprotein of vesicular stomatitis virus (VSVG) is usually used as the lentiviral vector envelope, but it has a wide spectrum of cell tropism which limits its application in vivo.…”

Immunization with lentiviral vector‑modified dendritic cells encoding ubiquitinated hepatitis B core antigen promotes Th1 differentiation and antiviral immunity by enhancing p38 MAPK and JNK activation in HBV transgenic mice

“…Our previous study demonstrated that LV-Ub-HBcAg can induce DC maturation and improve DC function in vitro (16). Our previous studies have shown that the production of Ub-HBcAg can promote HBcAg degradation into antigenic peptides by the UPS (16,19). These peptides are presented by DCs and are readily recognized by CD8 + T cells, whereas the activation of HBV-specific CD8 + T cells is critical for HBV control (36,37).…”

“…Ub-HBcAg and/or HBcAg were constructed as described previously (19). The lentiviral particles (LV-Ub-HBcAg and LV-HBcAg or LV) were prepared in 293T cells (American Type Culture Collection, Manassas, VA, USA) and tittered as described previously (19).…”

“…Ub-HBcAg and/or HBcAg were constructed as described previously (19). The lentiviral particles (LV-Ub-HBcAg and LV-HBcAg or LV) were prepared in 293T cells (American Type Culture Collection, Manassas, VA, USA) and tittered as described previously (19). Briefly, the recombinant pLOV.UBC.Ub-HBcAg.EGFP.3FLAG vector plasmid was constructed by inserting the Ub-HBcAg fragment into the BamHI and NheI sites of the pLOV.UBC.EGFP.3FLAG plasmid (Obio Technology Corp., Ltd., Shanghai, China).…”

“…Murine DCs were generated according to the protocol described by Chen et al (16). The DCs were transduced by the LVs (LV-Ub-HBcAg, LV-HBcAg, or LV) at a multiplicity of infection of 20 as described in our previous study (19).…”