The virus-like particle(VLPs) vaccine is an ideal HIV-1 vaccine, which can simultaneously induce a neutralizing antibody reaction and cell-mediated immunity effectively. In this study, two kinds of plasmids have been used, one can express the HIV-1 main structure proteins, Gagpol and Env, and the other contains an antibiotic gene. The two kinds of plasmids have been cotransfected into 293 cells. A stable cell line that can express Gagpol and Env proteins efficiently and lastingly has been screened. It has been confirmed that Gagpol and Env proteins in the cell culture supernatant can be self-assembled into virus-like particles. The authors have detected the secretion of VLPs in the cell medium, defined the peak of the secretion, and followed and monitored the stability of expression.
The induction of human immunodeficiency virus(HIV)-specific T-cell response is generally considered as critical to the development of effective immunity to HIV type 1(HIV-1). Recombinant Avipoxvirus vectors are used widely for vaccination against HIV-1, where the induction of a cytotoxic CD8+T-cell(CTL) response seems to be an important component of protective immunity. A recombinant fowlpox virus(rFPV/Gag-pol) expressing the Gag-pol protein of HIV was constructed and characterized. The specific expression protein in CEF cells infected by recombinant fowlpox and the specific antibody in the sera of mice immunized with rFPV were analyzed via Western-blot.
TENG Hong-gang LUE Shuai-ran LIU Da-wei JIANG Chun-lai ZHANG Xi-zhen YU Xiang-hui KONG Wei
Highly attenuated modified vaccinia Ankara(MVA) is sensitive to repeat freeze-thaw cycle and easy to lose activity. In order to make the activity of MVA vaccine remain stable during its manufacturing, storage, and administration, the lyophilization as a good option could be resorted to; through screening, the right stabilizer composition and its production procedure were obtained. The final moisture content of freezing-dried recombinant MVA-HIV vaccine was lower than 3%. It can be reconstituted quickly and shows regular physical appearance and stable potency. In vivo functional experiment, mice were divided randomly into the liquid vaccination group, the lyophilized vaccination group, and the control group. Having been DNA vaccine priming, the mice were boosted with a dose of 107 pfu MVA-HIV vaccine, which produced indistinguishable antibody titer and cytotoxic T-lymphocyte(CTL) level compared with those of liquid vaccination group(P>0.05). These results demonstrate that lyophilized MVA vaccine can induce high immunogenicity in mice.
ZHANG Yi-zhe JIANG Chun-lai YU Xiang-hui LOU Chao-ping ZHAO Dong-hai WU Yong-ge JIN Ying-hua LIU Cheng-shan KONG Wei
The rapid growth of the global HIV/AIDS epidemic makes it a high priority to develop an effective vaccine.Since a live attenuated or inactivated HIV vaccine is not likely to be approved for clinical application due to safety concerns,HIV virus like particles(VLPs) offer an attractive alternative because they are considered safer since they lack viral genome.We got a stable eukaryotic cell line by G418 resistance selection,engineered to express the HIV-1 structure protein Gag and Env efficiently and stably.We confirmed the presence of Gag and Env proteins in the cell culture supernatant and that they could self-assemble into VLPs.These VLPs were found to be able to elicit specific humoral and cellular immune response after immunization without any adjuvant.
