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Plant-produced Ebola Immune Complex as an Ebola Vaccine Candidate.

机译：植物产生的埃博拉免疫复合物，作为埃博拉疫苗的候选者。

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摘要

Ebola hemorrhagic fever (EHF) is a severe and often fatal disease in human and nonhuman primates, caused by the Ebola virus. Approximately 30 years after the first epidemic, there is no vaccine or therapeutic medication approved to counter the Ebola virus.;In this dissertation, a geminiviral replicon system was used to produce Ebola immune complex (EIC) in plant leaves and tested it as an Ebola vaccine. The EIC was produced in Nicotiana benthamiana leaves by fusing Ebola virus glycoprotein (GP1) to the C-terminus of heavy chain of 6D8 monoclonal antibody (mAb), which is specific to the 6D8 epitope of GP1, and co-expressing the fusion with the light chain of 6D8 mAb. EIC was purified by ammonium sulfate precipitation and protein A or protein G affinity chromatography. EIC was shown to be immunogenic in mice, but the level of antibody against Ebola virus was not sufficient to protect the mice from lethal the Ebola challenge.;Hence, different adjuvants were tested in order to improve the immunogenicity of the EIC. Among several adjuvants that we used, Poly(I:C), which is a synthetic analog of double-stranded ribonucleic acid that can interact with a Toll-like receptor 3, strongly increased the efficacy of our Ebola vaccine. The mice immunized with EIC co-administered with Poly(I:C) produced high levels of neutralizing anti-Ebola IgG, and 80% of the mice were protected from the lethal Ebola virus challenge. Moreover, the EIC induced a predominant T-helper type 1 (Th1) response, whereas Poly(I:C) co-delivered with the EIC stimulated a mixed Th1/Th2 response. This result suggests that the protection against lethal Ebola challenge requires both Th1 and Th2 responses.;In conclusion, this study demonstrated that the plant-produced EIC co-delivered with Poly(I:C) induced strong and protective immune responses to the Ebola virus in mice. These results support plantproduced EIC as a good vaccine candidate against the Ebola virus. It should be pursued further in primate studies, and eventually in clinical trials.
机译：埃博拉出血热（EHF）是人类和非人类灵长类动物中的一种严重且通常是致命的疾病，由埃博拉病毒引起。首次流行后约30年，尚无批准用于抵抗埃博拉病毒的疫苗或治疗药物。;本文使用双生病毒复制系统在植物叶片中产生埃博拉免疫复合物（EIC）并将其作为埃博拉病毒进行了测试疫苗。通过将埃博拉病毒糖蛋白（GP1）融合到6D8单克隆抗体（mAb）的重链C末端（对GP1的6D8表​​位具有特异性），并与融合蛋白共表达融合蛋白，从而在本氏烟草叶片中产生EIC。 6D8 mAb轻链。 EIC通过硫酸铵沉淀和蛋白A或蛋白G亲和色谱纯化。 EIC在小鼠中被证明具有免疫原性，但是抗埃博拉病毒的抗体水平不足以保护小鼠免受致命的埃博拉病毒攻击。因此，为了提高EIC的免疫原性，测试了各种佐剂。在我们使用的几种佐剂中，聚（I：C）是可以与Toll样受体3相互作用的双链核糖核酸的合成类似物，大大提高了埃博拉疫苗的功效。用EIC免疫与Poly（I：C）共同免疫的小鼠产生了高水平的中和性抗埃博拉IgG，并且80％的小鼠免受致命的埃博拉病毒攻击。此外，EIC诱导了主要的1型T辅助反应（Th1），而与EIC共递送的Poly（I：C）则刺激了混合的Th1 / Th2反应。该结果表明，对致命埃博拉病毒的攻击需要Th1和Th2响应。总之，这项研究表明，与Poly（I：C）共同提供的植物产生的EIC诱导了对埃博拉病毒的强力和保护性免疫反应。在小鼠中。这些结果支持植物产生的EIC作为抗埃博拉病毒的良好疫苗候选者。在灵长类动物研究中，甚至在临床试验中，都应进一步进行研究。

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  • 作者

    Phoolcharoen, Waranyoo.;

  • 作者单位

    Arizona State University.;

  • 授予单位 Arizona State University.;
  • 学科 Biology Botany.;Health Sciences Immunology.;Biology Molecular.
  • 学位 Ph.D.
  • 年度 2010
  • 页码 228 p.
  • 总页数 228
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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