Pre-clinical evaluation of a Zika virus vaccine

摘要 自2015年5月首次在美洲发现后，蚊子传播的寨卡病毒从巴西迅速传播， ZIKV病毒引起了全球的关注。巴西的ZIKV疫情也与一种 婴儿出生时患有小头畸形和神经系统疾病（如格林-巴利综合征）的人数显著增加， Barré综合征已被世界卫生组织宣布为“全球紧急情况”。寨卡病毒 与登革热、黄热病、日本脑炎和西尼罗河病毒有关，所有这些病毒都是节肢动物传播的 黄病毒基于文献中报道的成功的黄病毒疫苗方法作为初步尝试 为了开发寨卡病毒疫苗，我们已经产生了一组重组寨卡病毒亚单位疫苗， 在ZIKV包膜基因的胞外部分或ZIKV包膜基因的EDIII结构域上。的 黄病毒包膜基因的EDIII环是受体结合结构域，并且已被证明是最重要的受体结合结构域。 有希望成为抗病毒中和免疫的靶点。这两种抗原（E和EDIII）均产生为 单体（ZIKV-rE和ZIKV-rEDIII），作为与人IgG 1的fc部分融合的二聚体（ZIKV-rEhIg 和ZIKV-rEDIIIhIg），以及作为与T4纤维蛋白折叠子三聚化结构域融合的三聚体（ZIKV-rEfl和ZIKV-rEDIIIhIg）。 ZIKV-rEDIII fl）。在本文呈现的初步研究中，我们测试了三聚体ZIKV-rEfl的免疫原性。 和使用可溶性微针阵列（MNA）递送系统经皮递送的ZIKV-rEDIII fl， C57 BL 6小鼠。选择基于MNA的抗原递送系统是合理的，因为它的功效和它是 简单且易于适用于低卫生和纯社会经济环境。我们比较了 免疫原性与用腺病毒5基疫苗递送的相同抗原的免疫原性 vector.我们现在提议将测试扩展到产生的所有六种ZIKV重组亚单位疫苗。 这六种亚单位疫苗，命名为ZIKV-rE、ZIKV-rEDIII、ZIKV-rEhIg、ZIKV-rEDIII hIg、 将在C57 B6小鼠中比较经由MNA递送的ZIKV-rEfl和ZIKV-rEDIII fl。曾经表现最好的 选择ZIKV亚单位疫苗，评价佐剂的有益效果。最终，我们将测试 图10显示了所选亚单位疫苗在ZIKV替代动物攻击模型中的功效。我们假设 在初免/加强方案中施用ZIKV亚单位疫苗将引发ZIKV特异性免疫，其将导致ZIKV免疫应答。 有效地免疫孕鼠，并将被动免疫传递给新生儿，这将是 受ZIKV挑战的保护。

Abstract Since first detected in the Americas in May 2015 after rapidly spreading from Brazil, the mosquito-borne Zika virus (ZIKV) has attracted global attention. The ZIKV outbreak in Brazil has also been associated with a significant rise in the number of babies born with microcephaly and neurological disorders such as Guillain- Barré syndrome and has been declared a “global emergency” by the World Health Organization. Zika virus is related to dengue, yellow fever, Japanese encephalitis, and West Nile viruses, all of which are arthropod-borne flaviviruses. Building on successful flavivirus vaccine approaches reported in the literature as an initial attempt to develop a Zika virus vaccine, we have generated a panel of recombinant Zika virus subunit vaccines based on the extracellular portion of the ZIKV envelope gene or the EDIII domain of the ZIKV envelope gene. The EDIII loop of the flavivirus envelope gene is the receptor-binding domain and has been shown to be the most promising target for an antiviral neutralizing immunity. Both these antigens (E and EDIII) were generated as monomers (ZIKV-rE and ZIKV-rEDIII), as dimers in fusion with the fc portion of the human IgG1 (ZIKV-rEhIg and ZIKV-rEDIIIhIg), and as trimers in fusion with the T4 fibritin foldon trimerization domain (ZIKV-rEfl and ZIKV-rEDIIIfl). In the preliminary studies presented here, we tested the immunogenicity of trimeric ZIKV-rEfl and ZIKV-rEDIIIfl delivered transcutaneously using dissoluble microneedle array (MNA) delivery system into C57BL6 mice. The choice of the MNA-based antigen delivery system is justified because its efficacy and it is simple and easily adaptable to use in low hygiene and pure socio-economic environment. We compared their immunogenicity with the immunogenicity of the same antigens delivered with adenovirus 5-based vaccine vector. We now are proposing extending the testing to all six ZIKV recombinant subunit vaccines generated. The immunogenicity of these six subunit vaccines, named ZIKV-rE, ZIKV-rEDIII, ZIKV-rEhIg, ZIKV-rEDIIIhIg, ZIKV-rEfl, and ZIKV-rEDIIIfl, delivered via MNA, will be compared in C57B6 mice. Once the best performing ZIKV subunit vaccine is selected, the beneficial effect of adjuvant will be evaluated. Ultimately, we will test the efficacy of the selected subunit vaccine in a ZIKV surrogate animal challenge model. We hypothesize that administration of ZIKV subunit vaccines in a prime/boost regimen will elicit ZIKV-specific immunity that will lead to the effective immunization of pregnant mice with passage of passive immunity to newborns, which will be protected by ZIKV challenge.