Abstract

Ebola virus is the primary causative agent of viral hemorrhagic fever that is an epidemic disease and responsible for the massive premature deaths in humans. Despite knowing the molecular mechanism of its pathogenesis, to date, no commercial or FDA approved multiepitope vaccine is available against Ebola infection. The current study focuses on designing a multi-epitope subunit vaccine for Ebola using a novel immunoinformatic approach. The best predicted antigenic epitopes of Cytotoxic-T cell (CTL), Helper-T cells (HTL), and B-cell epitopes (BCL) joined by various linkers were selected for the multi-epitope vaccine designing. For the enhanced immune response, two adjuvants were also added to the construct. Further analysis showed the vaccine to be immunogenic and non-allergenic, forming a stable and energetically favorable structure. The stability of the unbound vaccine construct and vaccine/TLR4 was elucidated via atomistic molecular dynamics simulations. The binding free energy analysis (ΔG_Bind = −194.2 ± 0.5 kcal/mol) via the molecular mechanics Poisson-Boltzmann docking scheme revealed a strong association and thus can initiate the maximal immune response. Next, for the optimal expression of the vaccine construct, its gene construct was cloned in the pET28a + vector system. In summary, the Ebola viral proteome was screened to identify the most potential HTLs, CTLs, and BCL epitopes. Along with various linkers and adjuvants, a multi-epitope vaccine is constructed that showed a high binding affinity with the immune receptor, TLR4. Thus, the current study provides a highly immunogenic multi-epitope subunit vaccine construct that may induce humoral and cellular immune responses against the Ebola infection.

Communicated by Ramaswamy H. Sarma

摘要

埃博拉病毒是病毒性出血热的主要病原体，病毒性出血热是一种流行病，导致人类大量过早死亡。尽管知道其发病机制的分子机制，但迄今为止，还没有商业或 FDA 批准的多表位疫苗可用于对抗埃博拉病毒感染。目前的研究重点是使用一种新的免疫信息学方法为埃博拉病毒设计一种多表位亚单位疫苗。选择通过各种接头连接的细胞毒性T细胞（CTL）、辅助T细胞（HTL）和B细胞表位（BCL）的最佳预测抗原表位用于多表位疫苗设计。为了增强免疫反应，还向构建体中添加了两种佐剂。进一步的分析表明，该疫苗具有免疫原性和非过敏性，形成了一个稳定且充满活力的结构。通过原子分子动力学模拟阐明了未结合的疫苗构建体和疫苗/TLR4 的稳定性。结合自由能分析（ΔG _Bind = -194.2 ± 0.5 kcal/mol）通过分子力学 Poisson-Boltzmann 对接方案显示出强烈的关联，因此可以引发最大的免疫反应。接下来，为了疫苗构建体的最佳表达，将其基因构建体克隆到 pET28a + 载体系统中。总之，对埃博拉病毒蛋白质组进行了筛选，以确定最具潜力的 HTL、CTL 和 BCL 表位。连同各种接头和佐剂，构建了一种多表位疫苗，该疫苗与免疫受体 TLR4 具有高结合亲和力。因此，目前的研究提供了一种高度免疫原性的多表位亚单位疫苗构建体，可以诱导针对埃博拉病毒感染的体液和细胞免疫反应。

由 Ramaswamy H. Sarma 传达