Department of Microbiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, PR China.
Mol Immunol. 2013 Jun;54(2):109-14. doi: 10.1016/j.molimm.2012.11.007. Epub 2012 Dec 25.
Public health is still seriously threatened by dengue virus (DENV) and no vaccine against DENV is yet available for clinical use till now. In this study, DNA vaccine candidates encoding DENV serotype 2 (DENV-2) prM/E (premembrane and envelope proteins) and NS1 (non-structural 1 protein) with or without a gene adjuvant, granulocyte-macrophage colony-stimulating factor (GM-CSF), were evaluated in the aspects of immunity and protective efficacy in mice. We constructed three plasmids, pCAG-prM/E (which only expressed DENV2 prM/E), pCAG-prM/E/NS1 (which only expressed DENV2 prM/E/NS1) and pCAG-DG (which co-expressed DENV2 prM/E/NS1 and GM-CSF). The expressions of the recombined plasmids were analyzed by immuno-staining in Vero cells. Antibody responses and neutralization activity of the sera from the mice were assayed by ELISA and plaque reduction neutralization test after immunization with the plasmids. Immunized BALB/c mice were intracerebrally challenged with DENV2 to evaluate protective efficacy of the plasmids. The recombinant plasmids could be efficiently expressed in Vero cells and induced different levels of specific anti-DENV2 immune responses. The immunized mice were partially protected. The highest survival rate was observed in the pCAG-DG group although the anti-DENV2 titer and neutralization antibody titer were not the highest among the three groups. Our data suggested that pCAG-DG offered better protection against DENV2 infection.
公共卫生仍然受到登革病毒(DENV)的严重威胁,到目前为止,还没有针对 DENV 的疫苗可供临床使用。在这项研究中,我们评估了含有或不含有基因佐剂粒细胞-巨噬细胞集落刺激因子(GM-CSF)的 DENV 血清型 2(DENV-2)prM/E(前膜和包膜蛋白)和 NS1(非结构蛋白 1)的 DNA 疫苗候选物在小鼠中的免疫原性和保护效果。我们构建了三个质粒,pCAG-prM/E(仅表达 DENV2 prM/E)、pCAG-prM/E/NS1(仅表达 DENV2 prM/E/NS1)和 pCAG-DG(共表达 DENV2 prM/E/NS1 和 GM-CSF)。通过免疫染色在 Vero 细胞中分析重组质粒的表达。通过 ELISA 和噬斑减少中和试验测定免疫后小鼠血清的抗体反应和中和活性。用质粒免疫 BALB/c 小鼠后,通过脑内攻毒评估质粒的保护效果。重组质粒可在 Vero 细胞中高效表达,并诱导不同水平的特异性抗 DENV2 免疫反应。免疫小鼠得到部分保护。虽然 pCAG-DG 组的抗 DENV2 滴度和中和抗体滴度不是三组中最高的,但观察到最高的存活率。我们的数据表明,pCAG-DG 提供了针对 DENV2 感染的更好保护。