Establishment of cellular immunity of enhanced hepatitis B vaccine / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish the method to detect the cellular immune response of enhanced hepatitis B vaccine and make verification preliminary.</p><p><b>METHODS</b>Immunized BALB/c mice with enhanced hepatitis B vaccine and detected the IFN-gamma spots forming cells (SFC) of mouse spleen cell by Elispot. Optimized the conditions of the experiment. Cellular immune response between enhanced hepatitis B vaccine and normal hepatitis B vaccine by Elispot were compared.</p><p><b>RESULTS</b>IFN-gamma SFC was higher in 5microg dose than in 2microg dose after immunization with enhanced hepatitis B vaccine and IFN-gamma SFC was declined after immunization 3 weeks ago. IFN-gamma SFC was higher in stimulus by peptide than by protein. Compared to normal hepatitis B vaccine, IFN-gamma SFC was higher in enhanced hepatitis B vaccine.</p><p><b>CONCLUSION</b>Established the detection method to evaluate the cellular immunity of enhanced hepatitis B vaccine and tested the repeatability.</p>

Establishment and optimization of detection of the hepatitis B immune complexes / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish and optimize methods to detect the immune complexes (IC) of hepatitis B virus directly.</p><p><b>METHODS</b>A C1q solid phase ELISA, mouse anti-HBs MAb solid phase ELISA and the complement consumption assay were established to detect the IC and these methods were optimized.</p><p><b>RESULTS</b>All the three methods were highly sensitive, specific and reproducible. The C1q used for coating tended to lose its activity easily at room temperature. Although strict requirements are needed for the raw and processed materials for complement consumption assay and the process of manipulation is complex, it can quantitatively detect IC. Comparing to the C1q solid ELISA and complement consumption assay, the mouse anti-HBs MAb solid phase ELISA has its own merits: convenience and stability.</p><p><b>CONCLUSION</b>Mouse anti-HBs MAb solid phase ELISA is the best way to detect IC directly.</p>

Immune response for phase I clinical trial of a hepatitis B immunogenic complex therapeutic vaccine, YIC / 中华肝脏病杂志

<p><b>OBJECTIVE</b>A hepatitis B immunogenic complex therapeutic vaccine, yeast-derived recombinant HBsAg combined with human anti-HBs immunoglobulin (YIC), was evaluated for safety and immune response in phase I clinical trial.</p><p><b>METHODS</b>The subtypes IgG1, IgG2, IgG3 and IgG4 of serum anti-HBs collected from 20 immunized subjects were analyzed by ELISA. The lymphocyte proliferation assay was carried out in five subjects and was analyzed by 3H-thymidine incorporation. The assays for IFNgamma, IL-2, IL-4, IL-6, IL-10 and TNFalpha were measured using Human Cytometric Bead Array Kit with FACSCalibur.</p><p><b>RESULTS</b>The results showed that the subtypes of anti-HBs antibodies induced by 30, 60 and 90 microg YIC-immunized groups among all of the adult volunteers (20/20) were IgG1 and IgG3. The level of IgG1 was higher than that of IgG3 in each volunteer but the strength was different from each other. The rHBsAg-stimulated lymphocyte proliferation induced by three injections of 90 microg of YIC showed that the stimulation index was more than 2.0 in four out of the five individuals (4/5), ranging from 2.70 to 4.75. PHA-stimulated lymphocyte proliferation was not related to rHBsAg-stimulated lymphocyte proliferation. In the 60 microg YIC-immunized group there was no significant difference between the levels of IFNgamma, IL-2, IL-4, IL-6 and IL-10 at day 0 and day 42. At day 71, in comparison to day 0, the level of IFNgamma was higher in all eight subjects studied (P = 0.015) and the level of IL-2 was also increased in seven out of eight subjects (P = 0.002). In contrast, the levels of IL-4, IL-6, IL-10 and TNFalpha showed no significant difference in all the subjects (P-values: 0.298, 0.976, 0.202 and 0.996).</p><p><b>CONCLUSION</b>Our results indicate that this hepatitis B immunogenic complex therapeutic vaccine (YIC) can induce a potent anti-HBs response.</p>