Eliciting cross-neutralizing antibodies in mice challenged with a dengue virus envelope domain III expressed in Escherichia coli.
Eliciting cross-neutralizing antibodies in mice challenged with a dengue virus envelope domain III expressed in Escherichia coli.
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DOI：
10.1139/w11-137
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发表时间：
2012-03
影响因子：
2.8
通讯作者：
Jie Yang;Jun-lei Zhang;Wei Chen;Zhen Hu;Junmin Zhu;X. Fang;Wenchang Yuan;Ming Li;Xiaomei Hu-Xiaomei
中科院分区：
文献类型：
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作者：
Jie Yang;Jun-lei Zhang;Wei Chen;Zhen Hu;Junmin Zhu;X. Fang;Wenchang Yuan;Ming Li;Xiaomei Hu-Xiaomei
登革病毒（DENV）是蚊媒传染病病原体，对全球公共卫生构成严重威胁，目前尚无有效的治疗方法或疫苗。选择合适的单位作为候选物是登革热亚单位疫苗开发的基础。本研究根据DENV-2 E蛋白的功能，特别是E蛋白的中和表位，选择DENV-2 E蛋白的结构域III（EDIII），通过N端与细菌前导序列（pelB）融合，C端与6×His标签融合，在大肠杆菌中表达。在使用Ni-NTA亲和和阳离子交换层析的两步纯化后，His标记的EDIII被纯化至高达98%的均一性。该重组EDIII能够在BALB/c和C57 BL/6小鼠中触发高水平的中和抗体。重组EDIII及其鼠抗体均保护Vero细胞免受DENV-2感染。有趣的是，重组EDIII提供至少部分针对DENV-1感染的交叉保护。此外，EDIII抗体能够保护乳鼠免受体内病毒攻击。这些数据表明，具有在所有4种DENV血清型中保守的中和表位的基于小EDIII蛋白的候选分子具有重要意义。
Dengue viruses (DENVs) are mosquito-borne infectious pathogens that pose a serious global public health threat, and at present, no therapy or effective vaccines are available. Choosing suitable units as candidates is fundamental for the development of a dengue subunit vaccine. Domain III of the DENV-2 E protein (EDIII) was chosen in the present study and expressed in Escherichia coli by N-terminal fusion to a bacterial leader (pelB), and C-terminal fusion with a 6×His tag based on the functions of DENV structure proteins, especially the neutralizing epitopes on the envelope E protein. After two-step purification using Ni-NTA affinity and cation-exchange chromatography, the His-tagged EDIII was purified up to 98% homogenicity. This recombinant EDIII was able to trigger high levels of neutralizing antibodies in both BALB/c and C57BL/6 mice. Both the recombinant EDIII and its murine antibodies protected Vero cells from DENV-2 infection. Interestingly, the recombinant EDIII provides at least partial cross-protection against DENV-1 infection. In addition, the EDIII antibodies were able to protect suckling mice from virus challenge in vivo. These data suggest that a candidate molecule based on the small EDIII protein, which has neutralizing epitopes conserved among all 4 DENV serotypes, has important implications.