Z Huang1, S Lu, N Liu. 1. First Affiliated Hospital of Nanjing Medical University.
Abstract
OBJECTIVE: To observe the specific immune responses in H-2d mice after DNA immunization of HBV core gene. METHODS: The DNA vaccine (pJW4303/HBc) was constructed by inserting HBV core gene fragment into the reading frame of plasmid pJW4303. Both methods of gene gun and intramuscular immunization were used in this experiment. Anti-HBc (IgG) and its isotypes (IgG1, IgG2a) in mice sera were detected by ELISA. HBcAg specific cytotoxic T lymphocyte (CTL) activity was measured by 51Cr release assay. RESULTS: The expression of HBcAg was confirmed by ELISA and Western-blot in 293T cells transfected with pJW4303/HBc. The end-point titers of serum anti-HBc in immunized mice were 1: 328,050 (gene gun method) and 1:109,350 (intramuscular method). In both groups of gene gun and intramuscular immunization IgG2a level was slightly higher than IgG1. HBcAg specific CTL activities were 51. 1% in gene gun group and 55.2% in intramuscular group. CONCLUSION: In this experiment the DNA vaccine of pJW4303/HBc shows strong antigenecity in both cellular and humoral immunity.
OBJECTIVE: To observe the specific immune responses in H-2d mice after DNA immunization of HBV core gene. METHODS: The DNA vaccine (pJW4303/HBc) was constructed by inserting HBV core gene fragment into the reading frame of plasmid pJW4303. Both methods of gene gun and intramuscular immunization were used in this experiment. Anti-HBc (IgG) and its isotypes (IgG1, IgG2a) in mice sera were detected by ELISA. HBcAg specific cytotoxic T lymphocyte (CTL) activity was measured by 51Cr release assay. RESULTS: The expression of HBcAg was confirmed by ELISA and Western-blot in 293T cells transfected with pJW4303/HBc. The end-point titers of serum anti-HBc in immunized mice were 1: 328,050 (gene gun method) and 1:109,350 (intramuscular method). In both groups of gene gun and intramuscular immunization IgG2a level was slightly higher than IgG1. HBcAg specific CTL activities were 51. 1% in gene gun group and 55.2% in intramuscular group. CONCLUSION: In this experiment the DNA vaccine of pJW4303/HBc shows strong antigenecity in both cellular and humoral immunity.