■蓝舌(BT)对畜牧业构成重大威胁,影响各种动物物种,造成巨大的经济损失。许多BT病毒(BTV)血清型的存在阻碍了控制工作,强调广谱疫苗的需要。
■在这项研究中,我们评估了BTV关键非结构(NS)蛋白中的保守氨基酸序列,并鉴定了大量高度保守的鼠和牛特异性MHCI类限制性(MHC-I)CD8+和MHC-II限制性CD4+表位.然后我们筛选了这些保守表位的抗原性,变应原性,毒性,和溶解度。利用这些表位,我们开发了以Toll样受体(TLR-4)激动剂为基础的广谱多表位疫苗.使用C-IMMSIM服务器在计算机中评估预测的促炎细胞因子应答。使用Robetta和GalaxyWEB服务器实现了结构建模和细化。最后,我们通过广泛的100纳秒分子动力学模拟评估了对接复合物的稳定性,然后考虑将疫苗用于密码子优化和计算机模拟克隆.
■我们在NS1和NS2蛋白中发现了许多符合这些标准的表位,并开发了硅广谱疫苗。免疫模拟研究表明,这些疫苗在接种组中诱导高水平的IFN-γ和IL-2。蛋白质-蛋白质对接分析证明了对TLR-4具有强结合亲和力的有希望的表位。对接的复合物是稳定的,具有最小的均方根偏差和均方根波动值。最后,模拟克隆质粒的GC含量较高,密码子适应指数>0.8,表明它们适合在原核系统中表达蛋白质疫苗。
■这些下一代疫苗设计很有前景,需要在湿实验室实验中进一步研究以评估其免疫原性。安全,和功效在家畜中的实际应用。我们的发现为开发一个全面的,广谱疫苗,可能彻底改变畜牧业的BT控制和预防策略。
UNASSIGNED: Bluetongue (BT) poses a significant threat to the livestock industry, affecting various animal species and resulting in substantial economic losses. The existence of numerous BT virus (BTV) serotypes has hindered control efforts, highlighting the need for broad-spectrum vaccines.
UNASSIGNED: In this study, we evaluated the conserved amino acid sequences within key non-structural (NS) proteins of BTV and identified numerous highly conserved murine- and bovine-specific MHC class I-restricted (MHC-I) CD8+ and MHC-II-restricted CD4+ epitopes. We then screened these conserved epitopes for antigenicity, allergenicity, toxicity, and solubility. Using these epitopes, we developed in silico-based broad-spectrum multiepitope vaccines with Toll-like receptor (TLR-4) agonists. The predicted proinflammatory cytokine response was assessed in silico using the C-IMMSIM server. Structural modeling and refinement were achieved using Robetta and GalaxyWEB servers. Finally, we assessed the stability of the docking complexes through extensive 100-nanosecond molecular dynamics simulations before considering the vaccines for codon optimization and in silico cloning.
UNASSIGNED: We found many epitopes that meet these criteria within NS1 and NS2 proteins and developed in silico broad-spectrum vaccines. The immune simulation studies revealed that these vaccines induce high levels of IFN-γ and IL-2 in the vaccinated groups. Protein-protein docking analysis demonstrated promising epitopes with strong binding affinities to TLR-4. The docked complexes were stable, with minimal Root Mean Square Deviation and Root Mean Square Fluctuation values. Finally, the in silico-cloned plasmids have high % of GC content with > 0.8 codon adaptation index, suggesting they are suitable for expressing the protein vaccines in prokaryotic system.
UNASSIGNED: These next-generation vaccine designs are promising and warrant further investigation in wet lab experiments to assess their immunogenicity, safety, and efficacy for practical application in livestock. Our findings offer a robust framework for developing a comprehensive, broad-spectrum vaccine, potentially revolutionizing BT control and prevention strategies in the livestock industry.