Adenovirus mediated expression of interleukin 12 regulating hepatitis C virus E2 gene immunization-induced immune response / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To observe the regulating effect of hepatitis C virus (HCV) envelop (E) 2 gene immunization-induced immune responses by adenovirus mediated interleukin 12 (IL-12).</p><p><b>METHODS</b>HCV E2 protein was expressed and purified from NIH 3T3 and then used as an antigen to detect antibodies against HCV E2. With 51Cr release, SP2/0 expressing HCV E2 was used as target cell to detect specific cytotoxic T lymphocytes (CTL) response; adenovirus recombined IL-12 was propagated by 293 cell. HCV E2 recombinant and adenovirus recombined IL-12 were injected into the quadriceps femoris muscles and abdominal cavities of 6-8 weeks old BALB/C mice. Sera were collected at 2, 3, and 4 weeks and detected for antibodies for E2. Spleen cells isolated at 4 weeks were analyzed for specific CTL response.</p><p><b>RESULTS</b>It was found that expression of IL-12 at an undetectable level did enhance HCV E2 gene immunization-induced CTL activity and there was no effect on its hormonal immune response.</p><p><b>CONCLUSION</b>Using adenovirus to express interleukin 12 was helpful for regulation of HCV E2 gene immunization-induced immune response. Combined HCV E2 and IL-12 can render a strong anti-HCV CTL activity and may be of use in the development of HCV gene vaccine in the future.</p>

Preparation and application of the monoclonal antibody against hepatoma-specific gamma-glutamyltransferase / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To prepare a monoclonal antibody against hepatoma-specific gamma-glutamyltransferase (GGT-II) and study it's application.</p><p><b>METHODS</b>Two Bal B/C mice were immunized with pure GGT-II, then their spleen cells were separated and fused to SP 2/0 myeloma cells so as to make hybridoma cell strain which could yield monoclonal antibody against GGT-II. And it's effect of binding GGT-II was detected by competitive inhibitory enzyme linked-immunosorbance assay (ELISA).</p><p><b>RESULTS</b>A mouse hybridoma cell strain which could steadily secrete the monoclonal antibody against GGT-II was obtained and named 2G4F6B2. This monoclonal antibody belonged to IgG1 subclass and was specific to GGT-II, without cross-reaction to GGT-II. The result of detecting human serum GGT-II by ELISA with the monoclonal antibody accorded with that by polyacrylamide gradient electrophoresis.</p><p><b>CONCLUSION</b>The monoclonal antibody against GGT-II prepared in this study has high specificity and can be applied in clinic to detect human serum GGT-II.</p>