The immune efficiency of a recombinant adenovirus type 5 with type 35 fiber containing HIV-1 gag gene （rAd5/F35-mod. gag） was investigated in BALB/c mice, in which the rAd5/F35-mod, gag was firstly identified with P...The immune efficiency of a recombinant adenovirus type 5 with type 35 fiber containing HIV-1 gag gene （rAd5/F35-mod. gag） was investigated in BALB/c mice, in which the rAd5/F35-mod, gag was firstly identified with PCR, then transfected to 293 cells and the in vitro expression level of Gag protein was determined by Western blotting and indirect immuno-fluorescent assay. Mice were immunized with intramuscular injections of rAd5/F35-mod, gag, rAd5-mod, gag or DNA and were boosted after 3 weeks. To test the effect of pre-existing anti-viral immunity on immunization, mice were also injected with Ad5- GFP vector and then immunized 4 and 7 weeks later with Ad5/F35-mod. gag vector. The P24-specific IgG antibody in sera of immunized mice was determined by ELISA and the specific cytotoxic T lymphocyte （CTL） response was assayed by intracellular cytokine staining. It was demonstrated that the rAd5/F35- rood. gag vector could express efficiently the HIV Gag protein in 293 cells in vitro and induce strong HIV- specific immune responses in vivo. The strongest CTL and serum IgG response occurred when mice were immunized twice with injection of rAd5/F35 alone, but the anti-Ad5 antibody after primary infection with adenovirus could inhibit the specific immune responses induced by rAd5/F35 vector. It is concluded that single immunization with recombinant adenovirus rAd5/F35-mod, gag can induce specific CTL and serum IgG antibody responses in mice, but the immunogenicity of rAd5/F35 is comparably weaker than that of rAd5.展开更多
文摘The immune efficiency of a recombinant adenovirus type 5 with type 35 fiber containing HIV-1 gag gene （rAd5/F35-mod. gag） was investigated in BALB/c mice, in which the rAd5/F35-mod, gag was firstly identified with PCR, then transfected to 293 cells and the in vitro expression level of Gag protein was determined by Western blotting and indirect immuno-fluorescent assay. Mice were immunized with intramuscular injections of rAd5/F35-mod, gag, rAd5-mod, gag or DNA and were boosted after 3 weeks. To test the effect of pre-existing anti-viral immunity on immunization, mice were also injected with Ad5- GFP vector and then immunized 4 and 7 weeks later with Ad5/F35-mod. gag vector. The P24-specific IgG antibody in sera of immunized mice was determined by ELISA and the specific cytotoxic T lymphocyte （CTL） response was assayed by intracellular cytokine staining. It was demonstrated that the rAd5/F35- rood. gag vector could express efficiently the HIV Gag protein in 293 cells in vitro and induce strong HIV- specific immune responses in vivo. The strongest CTL and serum IgG response occurred when mice were immunized twice with injection of rAd5/F35 alone, but the anti-Ad5 antibody after primary infection with adenovirus could inhibit the specific immune responses induced by rAd5/F35 vector. It is concluded that single immunization with recombinant adenovirus rAd5/F35-mod, gag can induce specific CTL and serum IgG antibody responses in mice, but the immunogenicity of rAd5/F35 is comparably weaker than that of rAd5.
