Institute of Biotechnology and Genetic Engineering, The University of Agriculture, Peshawar, Pakistan.
Department of Botany and Microbiology, College of Science, King Saud University, Riyadh, Saudi Arabia.
Front Immunol. 2022 Oct 13;13:1004804. doi: 10.3389/fimmu.2022.1004804. eCollection 2022.
The current global outbreak of monkeypox (MPX) disease, caused by Monkeypox virus (MPXV), has resulted in 16 thousand infection cases, five deaths, and has been declared a global health emergency of international concern by the World Health Organization. Given current challenges in the safety of existing vaccines, a vaccine to prevent MPX infection and/or onset of symptoms would significantly advance disease management. In this context, a multi-epitope-based vaccine could be a well-suited approach. Herein, we searched a publicly accessible database (Virus Pathogen Database and Analysis Resource) for MPXV immune epitopes from various antigens. We prioritized a group of epitopes (10 CD8+ T cells and four B-cell epitopes) using a computer-aided technique based on desirable immunological and physicochemical properties, sequence conservation criteria, and non-human homology. Three multi-epitope vaccines were constructed (MPXV-1-3) by fusing finalized epitopes with the aid of appropriate linkers and adjuvant (beta-defensin 3, 50S ribosomal protein L7/L12, and Heparin-binding hemagglutinin). Codon optimization and cloning in the pET28a (+) expression vector ensure the optimal expression of each construct in the system. Two and three-dimensional structures of the constructed vaccines were predicted and refined. The optimal binding mode of the construct with immune receptors [Toll-like receptors (TLR2, TLR3, and TLR4)] was explored by molecular docking, which revealed high docking energies of MPXV-1-TLR3 (-99.09 kcal/mol), MPXV-2-TLR3 (-98.68 kcal/mol), and MPXV-3-TLR2 (-85.22 kcal/mol). Conformational stability and energetically favourable binding of the vaccine-TLR2/3 complexes were assessed by performing molecular dynamics simulations and free energy calculations (Molecular Mechanics/Generalized Born Surface Area method). immune simulation suggested that innate, adaptive, and humoral responses will be elicited upon administration of such potent multi-epitope vaccine constructs. The vaccine constructs are antigenic, non-allergen, non-toxic, soluble, topographically exposed, and possess favourable physicochemical characteristics. These results may help experimental vaccinologists design a potent MPX vaccine.
当前由猴痘病毒（MPXV）引起的全球猴痘（MPX）疾病爆发已导致 1.6 万例感染病例，5 例死亡，并被世界卫生组织宣布为国际关注的全球卫生紧急事件。鉴于现有疫苗安全性方面的当前挑战，预防 MPX 感染和/或症状发作的疫苗将极大地促进疾病管理。在这种情况下，基于多表位的疫苗可能是一种合适的方法。在此，我们从各种抗原中搜索了公共可访问数据库（病毒病原体数据库和分析资源）中的 MPXV 免疫表位。我们使用基于理想的免疫学和物理化学特性、序列保守性标准和非人类同源性的计算机辅助技术，对一组表位（10 个 CD8+T 细胞和 4 个 B 细胞表位）进行了优先级排序。通过使用适当的接头和佐剂（β-防御素 3、50S 核糖体蛋白 L7/L12 和肝素结合血凝素）融合最终确定的表位，构建了三种多表位疫苗（MPXV-1-3）。密码子优化和在 pET28a（+）表达载体中的克隆确保了每个构建体在该系统中的最佳表达。构建疫苗的二维和三维结构进行了预测和优化。通过分子对接探索了构建物与免疫受体[Toll 样受体（TLR2、TLR3 和 TLR4）]的最佳结合模式，结果表明 MPXV-1-TLR3（-99.09 kcal/mol）、MPXV-2-TLR3（-98.68 kcal/mol）和 MPXV-3-TLR2（-85.22 kcal/mol）的对接能高。通过进行分子动力学模拟和自由能计算（分子力学/广义 Born 表面积方法）评估了疫苗-TLR2/3 复合物的构象稳定性和能量有利结合。免疫模拟表明，给予这种有效的多表位疫苗构建物后，将引发先天、适应性和体液免疫反应。疫苗构建物具有抗原性、非变应原性、无毒、可溶性、拓扑暴露和具有良好的物理化学特性。这些结果可能有助于实验疫苗学家设计有效的 MPX 疫苗。
