In recent years, Hepatitis B virus (HBV) persistent infection mouse model with recombinant adeno-associated virus 8 carrying 1.3 copies of HBV genome (rAAV8-1.3HBV) is concerned. We studied and compared the efficacy among HBV persistent infection mice models by other serotypes except AAV8. First, we prepared and purified five viruses: rAAV1-1.3HBV, rAAV2-1.3HBV, rAAV5-1.3HBV, rAAV8-1.3HBV and rAAV9-1.3HBV. Then we injected each virus into 3 C57BL/6J mice with the dose of lx 1011 vg (Viral genome, vg) per mouse. We detected HBsAg and HBeAg in sera by enzyme-linked immunosorbent assay (ELISA) at different time points post injection. We killed mice 8 weeks post injection and took blood and livers for assay. We detected copies of HBV DNA by real-time quantitative PCR in sera and livers. Meantime, we detected HBcAg in the livers of mice by immunohistochemistry and further performed pathology analysis of these livers. The five groups of mice, HBeAg and HBsAg expression sustained 8 weeks in serological detection and HBV DNA was both detected in sera and livers at the time of 8 weeks post injection. HBeAg, HBsAg, HBV DNA copies expression levels in descending order were AAV8>AAV9>AAV1>AAV5>AAV2. HBcAg expression was detected in livers as well. Varied degrees of liver damage were shown in five groups of mice. This study provides more alternative AAV vector species to establish a persistent infection with hepatitis B model.
Animals ; Dependovirus ; classification ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Genome, Viral ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Mice ; Mice, Inbred C57BL ; Serogroup ; Virus Replication

<b>OBJECTIVEb>To study the quasispecies and mutation features of hepatitis B virus polymerase (HBV P) gene in chronic hepatitis B patients before and after lamivudine treatment.

<b>METHODSb>The HBV P gene was amplificated with PCR and cloned, then single strand conformation polymorphism / heteroduplex analysis (SSCP/HDA) was applied to analyze the quasispecies complexity and mutation characters of HBV P gene.

<b>RESULTSb>The quasispecies number of HBV P gene before treatment was larger than that after treatment (7 to 14 vs. 4 to 8, t = 3.98, P < 0.05). Six patients had one or two predominant quasispecies before therapy, but the percentages of predominant quasispecies were lower than those after therapy (33.3% to 81.8% vs. 78.8% to 90.9%, t = 3.42, P < 0.05). By sequencing the predominant clones, there were 2 patients with M550V/L528M mutation, 3 patients with M550I mutation and 1 patient with wild type after lamivudine therapy. Additionally, there were individualization mutations which had no obvious tender.

<b>CONCLUSIONb>Quasispecies of hepatitis B virus are changed under the selection of lamivudine, meanwhile, the mutation at YMDD motif emerges.

Antiviral Agents ; therapeutic use ; DNA, Viral ; metabolism ; DNA-Directed DNA Polymerase ; genetics ; Female ; Hepatitis B virus ; classification ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Male ; Point Mutation ; genetics

Vertical transmission from mother to child, the main route of chronic hepatitis B virus (HBV) infection in the East Asia, is considered one of the most important predictors for the response to antiviral therapies as well as its complications such as cirrhosis and hepatocellular carcinoma. Therefore, it is critical in both etiologic and prognostic aspects to confirm whether or not chronic HBV infection is acquired vertically. This study investigated whether mother-to-child infection could be proved by the phylogenetic analyses of HBV pre-S/S genes ever since several decades have elapsed in mother-child pairs with presumed vertical transmission. The pre-S and S regions of HBVs were compared and analyzed phylogenetically in a total of 36 adults (18 mother-child pairs) with chronic HBV infection. All of the isolates of HBV were genotype C and serotype adr. The divergence between mothers and offsprings was 0 to 1.5%. Phylogenetic trees revealed that 17 of 18 pairs (94%) with presumed vertical transmission were grouped into the same cluster. Vertical transmission from mother to child could be strongly suggested even in adults with a history of several decades of HBV infection using the phylogenetic analyses of pre-S and S genes.
Adult ; Aged ; DNA, Viral/analysis ; Female ; Genotype ; Hepatitis B Surface Antigens/classification/*genetics ; Hepatitis B virus/classification/*genetics/metabolism ; Hepatitis B, Chronic/diagnosis/*virology ; Humans ; Infectious Disease Transmission, Vertical ; Male ; Middle Aged ; Mothers ; Phylogeny ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Serotyping ; Young Adult