Lentiviral vector encoding ubiquitinated hepatitis B core antigen induces potent cellular immune responses and therapeutic immunity in HBV transgenic mice

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

“…To achieve the targeting lentiviral particles (LVDC-UbHBcAg-LIGHT), we transduced 80% confluent 293T cells with a combination of three elements: the appropriate H4725 (pLOV-UBC-UB-HBcAg-EGFP-P2A-Tnfsf14-3FLAG) backbone plasmid (10μg), the engineered envelope plasmid (5 μg), along with 5μg of the psPAX2 packaging plasmid using Lipofectamine 2000 (Invitrogen). Plasmid H4725 was assembled by cloning the EGFP-P2A-Tnfsf14-3FLAG gene into the BamHI/XbaI sites of the CN1043 (pLOV-UBC-Ub-HBcAg-EGFP-3FLAG) which was maintained in our lab [10,11]. We generated the targeting envelope plasmid (SVGmu) by alterations including an insertion of 10-amino acid sequence (MYPYDVPDYA) between amino acids 71 and 74 of the SVG E2, mutations of 157KE158 into 157AA158 of the SVG E2.…”

“…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

“…H-2K d HBV-transgenic BALB/c mice (half male and half female) were obtained from the Key Liver Army Laboratory of No.458 Hospital (Guangzhou, China). They were 6-8 weeks old (20-23 g) and their characteristics were as described previously (17,18). All mice were housed under specific pathogen-free conditions (22-24˚C; humidity 50-55%; 12 h light/dark cycle), with free access to food and water.…”

Peer Review #3 of "Application of built-in adjuvants for epitope-based vaccines (v0.1)"

“…In addition, the nanoscale structure of HBcAg can be more effectively identified and processed by APCs (Lee et al, 2009;Ong et al, 2017). Therefore, HBcAg has been used as an vaccine carrier for several exogenous pathogens (e.g., hepatitis B, C, and E virus, influenza virus, foot-and-mouth disease virus, Human enterovirus 71, coxsackievirus A16, and Chlamydia trachomatis), and the immunogenicity of recombinant HBc-based virus like particle (VLP) vaccines against pathogens has also been verified in animal models (Dai et al, 2016;Su et al, 2013;Zheng et al, 2016;Chu et al, 2016;Zhu et al, 2016;Wu et al, 2017;Jiang et al, 2017). VLPs are the self-assembled structural proteins of most viruses and can stimulate the immune response in the absence of an adjuvant by exposing pathogen epitopes and simulating the structure of natural viruses (Plummer & Manchester, 2011;Yang et al, 2016).…”

“…Manuscript to be reviewed immunogenicity of such epitopes (Dai et al, 2016;Reynolds et al, 2015). Recently, Wu et al developed a novel vaccine against chickenpox and hand-foot-mouth disease (HFMD) by constructing three VLPs with HBcAg used as a carrier protein, and epitopes derived from varicellazoster virus (VZV)-gE, EV71 (enterovirus71)-VP1, and EV71-VP2 were displayed by HBcAg .…”