Hepacivirus ; Hepatitis C ; genetics ; Humans ; MicroRNAs ; genetics

Genes, Viral ; genetics ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Recombination, Genetic

Genetic Heterogeneity ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; Replicon ; genetics

Enzyme-Linked Immunosorbent Assay ; Hepacivirus ; genetics ; Hepatitis C ; diagnosis ; drug therapy ; Hepatitis C Antibodies ; blood ; Humans

To evaluate the clinical feasibility of the antibody titer against a chimeric polypeptide (named Core 518), in which a domain of Core and NS3 of hepatitis C virus (HCV) was fused, ELISA was performed in a total of 76 serum samples. Each serum was serially diluted using two-fold dilution method with distilled water into 10 concentrations. They were all positive for second generation anti-HCV assay (HCV EIA II; Abbott Laboratories). Genotyping RT-PCR, quantitative competitive RT-PCR, and RIBA (Lucky Confirm; LG Biotech) were also assayed. Anti-Core 518 antibody was detected in x 12800 or higher dilutions of sera from 35 of 43 chronic hepatitis C (81.4%) and nine of 16 hepatocellular carcinoma sera (56.3%), one of four cirrhosis (25%), 0 of four acute hepatitis C, and one of nine healthy isolated anti-HCV-positive subjects (p=0.0000). The anti-Core 518 antibody titers were well correlated with the presence of HCV RNA in serum (p=0.002). The anti-Core 518 antibody titers decreased significantly in nine of ten responders to IFN-alpha treatment. Monitoring anti-Core 518 titers may be helpful not only for differentiating the status of HCV infection among patients with various type C viral liver diseases, but also for predicting responses to IFN-alpha treatment.
Adult ; Aged ; Female ; Genotype ; Hepatitis C/immunology* ; Hepatitis C/drug therapy* ; Hepatitis C/diagnosis ; Hepatitis C/blood ; Hepatitis C Antibodies/immunology* ; Hepatitis C Antibodies/blood ; Hepatitis C Antigens/immunology* ; Hepatitis C-Like Viruses/immunology* ; Hepatitis C-Like Viruses/genetics ; Human ; Immunoblotting ; Interferon Alfa-2a/therapeutic use* ; Male ; Middle Age ; RNA, Viral/blood ; Recombinant Fusion Proteins/immunology ; Viral Core Proteins/immunology* ; Viral Nonstructural Proteins/immunology*

Hepatitis C ; genetics ; prevention & control ; therapy ; Humans ; MicroRNAs

Animals ; Hepacivirus ; genetics ; Hepatitis C ; immunology ; prevention & control ; therapy ; virology ; Humans ; Mutation ; Viral Hepatitis Vaccines ; immunology

BACKGROUNDTo study preparation of polyvalent DNA vaccine and the control of multiple gene expression.

METHODSA bicistronic vector pcDNA3.0BA was constructed from pcDNA3.0. HCV PC154 gene and HBV preS2S gene were inserted into this vector to form bicistronic expression construct pcDNA3.0BAPC154S2S and monocistronic expression construct pcDNA3.0BAPC154 or pcDNA3.0BAS2S. These plasmids were transiently expressed in COS-7 cells and injected into muscles of BALB/c mice.

RESULTSpcDNA3.0BA contains two cistronic units, which can co-express two kinds of genes, with the first immunogen gene and the second gene serving as additional immunogen or as modulator for the immune responses. HBV surface Ag and HCV core Ag were coexpressed in vitro. The antibody responses and lymphoproliferation to antigens were similar between bicistronic and monocistronic expression construct in mice.

CONCLUSIONpcDNA3.0BA is a novel vector, which can coexpress two proteins and elicit polyvalent immune responses.

Animals ; COS Cells ; Cercopithecus aethiops ; DNA, Recombinant ; immunology ; Gene Expression ; Hepatitis B ; blood ; immunology ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis C ; blood ; immunology ; Hepatitis C Antibodies ; blood ; Hepatitis C Antigens ; genetics ; immunology ; Immunization ; methods ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Vaccines, DNA ; genetics ; immunology ; Viral Hepatitis Vaccines ; genetics ; immunology

Hepacivirus ; genetics ; Hepatitis C ; diagnosis ; genetics ; therapy ; Humans ; Interleukins ; genetics ; Polymorphism, Single Nucleotide ; Prognosis

Hepatitis C ; genetics ; Humans ; RNA, Catalytic ; genetics ; RNA, Small Nuclear ; genetics