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Hepatitis C virus (HCV) infection is still a serious global health burden. Despite improved therapeutic options, a preventative vaccine would be desirable especially in undeveloped countries. Traditionally, highly conserved epitopes are targets for antibody-based prophylactic vaccines. In HCV-infected patients, however, neutralizing antibodies are primarily directed against hypervariable region I (HVRI) in the envelope protein E2. HVRI is the most variable region of HCV, and this heterogeneity contributes to viral persistence and has thus far prevented the development of an effective HVRI-based vaccine. The primary goal of an antibody-based HCV vaccine should therefore be the induction of cross-reactive HVRI antibodies. In this study we approached this problem by presenting selected cross-reactive HVRI variants in a highly symmetric repeated array on capsid-like particles (CLPs). SplitCore CLPs, a novel particulate antigen presentation system derived from the HBV core protein, were used to deliberately manipulate the orientation of HVRI and therefore enable the presentation of conserved parts of HVRI. These HVRI-CLPs induced high titers of cross-reactive antibodies, including neutralizing antibodies. The combination of only four HVRI CLPs was sufficient to induce antibodies cross-reactive with 81 of 326 (24.8%) naturally occurring HVRI peptides. Most importantly, HVRI CLPs with AS03 as an adjuvant induced antibodies with a 10-fold increase in neutralizing capability. These antibodies were able to neutralize infectious HCVcc isolates and 4 of 19 (21%) patient-derived HCVpp isolates. Taken together, these results demonstrate that the induction of at least partially cross-neutralizing antibodies is possible. This approach might be useful for the development of a prophylactic HCV vaccine and should also be adaptable to other highly variable viruses.

Induction of hepatitis B virus (HBV)-specific cytotoxic T cells by therapeutic immunization may be a strategy to treat chronic hepatitis B. In the HBV animal model, woodchucks, the application of DNA vaccine expressing woodchuck hepatitis virus (WHV) core antigen (WHcAg) in combination with antivirals led to the prolonged control of viral replication. However, it became clear that the use of more potent vaccines is required to overcome WHV persistence. Therefore, we asked whether stronger and more functional T-cell responses could be achieved using the modified vaccines and an optimized prime-boost vaccination regimen. We developed a new DNA plasmid (pCGWHc) and recombinant adenoviruses (AdVs) showing high expression levels of WHcAg. Mice vaccinated with the improved plasmid pCGWHc elicited a stronger WHcAg-specific CD8(+) T-cell response than with the previously used vaccines. Using multicolor flow cytometry and an in vivo cytotoxicity assay, we showed that immunization in a DNA prime-AdV boost regimen resulted in an even more vigorous and functional T-cell response than immunization with the new plasmid alone. Immunization of naïve woodchucks with pCGWHc plasmid or AdVs induced a significant WHcAg-specific degranulation response prior to the challenge, this response had not been previously detected. Consistently, this response led to a rapid control of infection after the challenge. Our results demonstrate that high antigen expression levels and the DNA prime-AdV boost immunization improved the T-cell response in mice and induced significant T-cell responses in woodchucks. Therefore, this new vaccination strategy may be a candidate for a therapeutic vaccine against chronic HBV infection.

Background: Liver transplantation is often the ultimate option of therapy for chronically hepatitis B virus (HBV) infected patients. Prevention of reinfection is therapy intensive and cost-effective. Adoptive transfer of HBV-specific immunity with the liver from an immune living liver donor (LLD) could be a new approach to prevent reinfection., Methods: Forty-six potential LLDs were vaccinated against HBV. Humoral (antibodies to hepatitis B virus surface antigen [anti-HBs]-titer) and cellular (IFN-gamma-ELISpot and proliferation-assay) immune responses were examined in donors after immunization and in recipients before and after transplantation., Results: Anti-HBs-titers of up to 50,000 IU/L were detected in LLDs. Fourteen recipients received livers from these donors. We detected humoral immunity in one HBV-naïve recipient and in one chronically HBV-infected recipient after transplantation. A transfer of cellular immunity (SI>3) was seen in three recipients. These patients received livers from donors with high anti-HBs-titers of more than 9000 IU/L. Cellular immunity was also detected in the corresponding donors (SI >3 and spots >22)., Conclusions: Our study demonstrates that HBV-specific humoral and cellular immunity can be transferred by liver transplantation after vaccination of the donors. The transfer of B-cell and T-cell immunity correlates with the magnitude of immune responses in the donor.

Non-responsiveness to conventional hepatitis B vaccines in individuals at high risk of exposure to hepatitis B virus (HBV) is an important public health problem and of particular relevance in health care providers. Yeast-derived conventional HBsAg vaccines fail to induce protective antibody titers in up to 10% of immune competent vaccinees. Therefore, a third generation HBV vaccine, Sci-B-Vac™, was developed which contains in addition to the small S antigen the PreS1 and PreS2 antigens. This vaccine proved to induce a highly potent cellular and humoral immune response in healthy individuals as well as protective antibody levels in non- and low-responders to conventional HBV vaccines. The aim of the study was to examine whether Sci-B-Vac™ triggers cellular and humoral immunity in individuals who failed immunization with conventional vaccines. We immunized 21 volunteers (15 non- and 6 low-responders) according to the standard vaccination schedule (0, 4 and 24 weeks), determined the cellular immunity by proliferation assay and interferon (IFN)-γ ELISpot and measured the anti-HBs antibody titers prior to each vaccination and four weeks after the third vaccine dose. Following three vaccinations, PreS/S-specific T-cell proliferation was detected in 8 out of 15 non-responders and 5 out of 6 low-responders. Specific IFN-γ responses were measured in 2 out of 15 non-responders and 4 out of 6 low-responders. All but one (20/21) study participants developed anti-HBs titers ≥10 IU/l after three vaccinations. Anti-HBs ≥100 IU/L were detected in 12 out of 15 non-responders and in 6 out of 6 low-responders. Anti-HBs ≥10 IU/l and <100 IU/l were found in 2 non-responders. These results indicate that Sci-B-Vac™ induces cellular immunity as well as protective anti-HBs antibody titers in non- and low-responders. In conclusion, these results confirm that Sci-B-Vac™ should be administered to non-responders to conventional HBV vaccines and patients with impaired immune function. [ABSTRACT FROM AUTHOR]

Abstract: Woodchuck hepatitis virus (WHV) is a member of family Hepadnaviridae and closely related to hepatitis B virus (HBV). The WHV core protein (WHcAg) is a strongly immunogenic protein and forms virus-like particles. WHcAg may represent a suitable carrier system for B- and T-cell epitopes. However, the lack of a high expression system for WHcAg and defined antibodies to detect WHcAg prevents the use of this carrier system. In the present study, vectors expressing WHcAg with carboxyl-terminal truncations were constructed to determine the region of WHcAg required for assembly. The first 144 or 149 amino acid residues of WHcAg were able to efficiently assemble into particulate structures. Both truncated forms of WHcAg were accumulated in E. coli as uniform particles with a diameter of 34nm in large quantities and could be purified in milligram scale. As expected, the particles of truncated WHcAg retained the antigenicity of the full length WHcAg. However, denatured WHcAg remained to be reactive with specific antisera, suggesting that WHcAg may possess additional linear B-cell epitopes. Monoclonal antibodies against denatured WHcAg were generated and tested for their specificity. Five antibodies were found to direct the N-terminal region of WHcAg. Due to the conservation of the amino acid sequence in this region of WHcAg and HBcAg, these antibodies recognized recombinant HBcAg as well. Thus, this linear B-cell epitope is conserved on the core proteins of hepadnaviruses. [Copyright &y& Elsevier]