Identification of novel hepatitis B virus therapeutic vaccine candidates derived from polymerase protein.

Hepatitis B virus (HBV) infection is a worldwide health problem with high morbidity and mortality rates. The therapeutic vaccine is a promising method of treatment, and HBV polymerase plays a vital role in viral replication. Therefore, a therapeutic vaccine that binds to HBV DNA polymerase may control HBV infection. We predicted and selected epitopes of polymerase using online databases and analysis software. We then performed molecular docking and peptide binding assays to evaluate the binding energies and affinities between polymerase epitopes and the HLA-A0201 molecule. Finally, we induced T cells from the peripheral blood mononuclear cells (PBMCs) of healthy donors using each epitope and quantified the functions of epitope-specific T cells by IFN-γELISPOT assay, T2 cell cytotoxicity assay, HepG2.2.15 cell cytotoxicity assay and HBV gene expression assays. Four epitopes (RVTGGVFLV, GLLGFAAPF, LLDDEAGPL and YMDDVVLGA) had low binding energy and two epitopes (RVTGGVFLV and GLLGFAAPF) had a high binding affinity. The T cells stimulated by two epitopes (GLLGFAAPF and HLYSHPIIL) had a greater ability to induce immune response and suppress HBV. The HBV DNA polymerase epitopes identified in this study are promising targets for designing an epitope-based therapeutic vaccine against HBV.

Hepatitis B virus-specific effector CD8+ T cells are an important determinant of disease prognosis: A meta-analysis.

BACKGROUND: Hepatitis B virus (HBV)-specific effector CD8+ T cells are critical for viral clearance. To determine the effects of HBV-specific effector CD8+ T cells on HBV infection, we performed a meta-analysis of the available literature. METHODS: Electronic database searches identified appropriately designed studies that detected specific CD8+ T cells in HBV-infected patients. Our main endpoints were the course of infection, seroconversion of HBV "e" antigen (HBeAg), the level of HBVDNA, and alanine aminotransferase (ALT) activity. We used a fixed/random model for analysis, according to the results of a heterogeneity test (P value of Q-squared, I2). RESULTS: Our searches found five eligible articles. Pooled estimation of the reported results showed that levels of specific CD8+ T cells were significantly higher in patients with acute hepatitis B than in patients with chronic hepatitis B (odds ratio [OR] = 76.30, 95% confidence interval [CI]: 15.37-378.70). With respect to chronic hepatitis B, patients with <107 copies/ml HBVDNA had higher levels of specific CD8+ T cells relative to patients with >107 copies/ml HBVDNA, but the difference had no statistics significance (OR: 3.89, 95% CI: 0.71-21.33). Patients with negative HBeAg or positive anti-HBeAg antibody (anti-HBe) results had significantly higher levels of specific CD8+ T cells versus patients with positive HBeAg results (OR: 5.82, 95% CI: 1.41-24.13). There were no significant associations between the levels of specific CD8+ T cells and serum ALT activity (OR = 0.86, 95% CI: 0.01-74.15). CONCLUSION: HBV-specific effector CD8+ T cells influence the disease activity in HBV-infected patients in various ways and determine prognosis by eliminating the virus. Therefore, efforts of studying HBV-specific effector CD8+ T cells focused vaccine are potentially needed.

Analysis of epitope-based vaccine candidates against the E antigen of the hepatitis B virus based on the B genotype sequence: An in silico and in vitro approach.

Chronic hepatitis B virus infection is a worldwide health problem with no current effective strategy to achieve a cure. The Hepatitis B virus (HBV) E antigen (HBeAg) has a negative effect on the immune system and a therapeutic vaccine is a promising strategy in order to treat chronic virus infection. In this study, we analyzed and identified the MHC-I, MHC-II and B cell epitopes of the HBeAg based on a B genotype sequence of HBV using a bioinformatic approach and in vitro experiments. The computational approach provided us with four epitopes (LLWFHISCL, YLVSFGVWI, MQLFHLCLI, TVLEYLVSF) of the specific MHC-I allele HLA-A0201 that conformed to all criteria. Molecular docking and a peptide binding assay showed that epitope TVLEYLVSF had the lowest binding energy and epitope LLWFHISCL had the highest binding affinity to the HLA-A0201 molecule. An interferonγenzyme-linked immunospot assay and cytotoxicity assay revealed that epitope LLWFHISCL had the highest ability to induce and stimulate T cells. Furthermore, we determined four core peptides of MHC-II epitopes and a region of the B cell epitope. The epitopes and region identified in this research may be helpful in designing epitope-based vaccines and boosting the mechanism research of HBeAg and its effect on the immune system.

In Silico Analysis of Epitope-Based Vaccine Candidates against Hepatitis B Virus Polymerase Protein.

Hepatitis B virus (HBV) infection has persisted as a major public health problem due to the lack of an effective treatment for those chronically infected. Therapeutic vaccination holds promise, and targeting HBV polymerase is pivotal for viral eradication. In this research, a computational approach was employed to predict suitable HBV polymerase targeting multi-peptides for vaccine candidate selection. We then performed in-depth computational analysis to evaluate the predicted epitopes' immunogenicity, conservation, population coverage, and toxicity. Lastly, molecular docking and MHC-peptide complex stabilization assay were utilized to determine the binding energy and affinity of epitopes to the HLA-A0201 molecule. Criteria-based analysis provided four predicted epitopes, RVTGGVFLV, VSIPWTHKV, YMDDVVLGA and HLYSHPIIL. Assay results indicated the lowest binding energy and high affinity to the HLA-A0201 molecule for epitopes VSIPWTHKV and YMDDVVLGA and epitopes RVTGGVFLV and VSIPWTHKV, respectively. Regions 307 to 320 and 377 to 387 were considered to have the highest probability to be involved in B cell epitopes. The T cell and B cell epitopes identified in this study are promising targets for an epitope-focused, peptide-based HBV vaccine, and provide insight into HBV-induced immune response.