Pre-clinical evaluation of a Zika virus vaccine

摘要 自2015年5月从巴西迅速传播后首次在美洲发现以来，蚊媒寨卡病毒 病毒(ZIKV)已引起全球关注。巴西的ZIKV疫情也与 出生时患有小头畸形症和吉兰等神经系统疾病的婴儿数量显著增加- 巴雷综合征已被世界卫生组织宣布为“全球紧急状态”。寨卡病毒是 与登革热、黄热病、日本脑炎和西尼罗河病毒有关，所有这些病毒都是由节肢动物传播的 黄病毒。在文献中报道的成功的黄病毒疫苗方法的基础上进行初步尝试 为了开发寨卡病毒疫苗，我们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。这个 黄病毒囊膜基因的EDIII环是受体结合域，已被证明是最多的 有望成为抗病毒中和免疫的靶点。这两种抗原(E和EDIII)都是作为 单体(ZIKV-Re和ZIKV-rEDIII)作为二聚体与人IgG1的Fc部分融合(ZIKV-rEhIg 和ZIKV-rEDIIIhIg)，以及与T4纤维蛋白折叠三聚结构域融合的三聚体(ZIKV-refl和 ZIKV-rEDIIIfl)。在这里介绍的初步研究中，我们测试了三聚体zikv-refl的免疫原性。 和ZIKV-rEDIIIf1使用可溶性微针阵列(MNA)递送系统经皮递送到 C57BL6小鼠。选择基于MNA的抗原递送系统是合理的，因为它的有效性和 简单，易于适应在卫生条件差和纯社会经济环境中使用。我们比较了他们的 免疫原性与腺病毒5型疫苗所传递的相同抗原的免疫原性 向量。我们现在建议将测试扩大到所有六种ZIKV重组亚单位疫苗。 检测了ZIKV-Re、ZIKV-rEDIII、ZIKV-rEhIg、ZIKV-rEDIIIhIg六种亚单位疫苗的免疫原性。 通过MNA传递的ZIKV-ReFl和ZIKV-rEDIIIfl将在C57B6小鼠身上进行比较。曾经的最佳表演 选择ZIKV亚单位疫苗，对佐剂的有益效果进行评价。最终，我们将测试 所选亚单位疫苗在ZIKV代理动物挑战模型中的效力。我们假设 在基础/增强方案中接种ZIKV亚单位疫苗将诱导ZIKV特异性免疫，这将导致 通过对新生小鼠的被动免疫来有效免疫孕鼠，这将是 受到ZIKV挑战赛的保护。

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.