Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China.
Viral Immunol. 2013 Feb;26(1):68-74. doi: 10.1089/vim.2012.0073. Epub 2013 Jan 18.
DNA vaccine can induce the prolonged immune responses against the encoded antigen with the appropriate adjuvant. To study the immunogenicity of the HIV-1 vif DNA vaccine in inducing the humoral and cellular immune responses and the immunoadjuvant effect of LIGHT, which is a member of TNF superfamily and can stimulate the proliferation of naïve T cells as a co-stimulatory molecule, DNA vaccine plasmid pcDNA-Vif was constructed by inserting HIV-1 vif gene into the downstream of CMV promoter in eukaryotic expression vector pcDNA3.1(+). In vitro expression of HIV-1 Vif in pcDNA-Vif-transfected HeLa cells was confirmed in transcriptional and protein level by RT-PCR and Western blot, respectively. After BALB/c mice were injected muscularly with DNA vaccines for three times, the specific immune responses were analyzed. The data showed that anti-Vif antibody response, Vif-specific T cell proliferation, and CTL activities were induced in the mice that were inoculated with HIV-1 vif DNA vaccine plasmid. Interestingly, stronger humoral and cellular immune responses were detected in mice that were immunized with plasmid pcDNA-Vif and pcDNA-LIGHT together compared to the single immunization with plasmid pcDNA-Vif alone. Together, the results of the study suggest that candidate HIV-1 DNA vaccine can elicit HIV-1 Vif-specific immune responses in mice and that LIGHT plays the role of immunoadjuvant in co-immunization with DNA vaccine.
DNA 疫苗可以与适当的佐剂一起诱导针对编码抗原的长期免疫反应。为了研究 HIV-1 vif DNA 疫苗在诱导体液和细胞免疫反应中的免疫原性以及 LIGHT 的免疫佐剂作用,LIGHT 是 TNF 超家族的成员,可以作为共刺激分子刺激幼稚 T 细胞的增殖,构建了 HIV-1 vif 基因插入真核表达载体 pcDNA3.1(+)中 CMV 启动子下游的 pcDNA-Vif DNA 疫苗质粒。通过 RT-PCR 和 Western blot 分别在转录和蛋白水平上证实了 pcDNA-Vif 转染的 HeLa 细胞中 HIV-1 Vif 的体外表达。在 BALB/c 小鼠肌肉注射 DNA 疫苗 3 次后,分析了特异性免疫反应。结果表明,接种 HIV-1 vif DNA 疫苗质粒的小鼠诱导了抗 Vif 抗体反应、Vif 特异性 T 细胞增殖和 CTL 活性。有趣的是,与单独接种 pcDNA-Vif 质粒相比,同时接种 pcDNA-Vif 和 pcDNA-LIGHT 质粒的小鼠中检测到更强的体液和细胞免疫反应。总之,该研究结果表明候选 HIV-1 DNA 疫苗可以在小鼠中引发 HIV-1 Vif 特异性免疫反应,并且 LIGHT 在与 DNA 疫苗共同免疫中发挥免疫佐剂作用。