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INTRODUCTION

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.

METHODOLOGY

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.

RESULTS

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.

DISCUSSION

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.

INTRODUCTION

METHODOLOGY

RESULTS

DISCUSSION

简介

蓝舌病（BT）对畜牧业构成重大威胁，影响多种动物物种，并导致重大经济损失。存在许多 BTV 血清型阻碍了控制工作，突出了对广谱疫苗的需求。

方法

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，并鉴定了许多高度保守的牛和猪 MHC I 限制性（MHC-I）CD8+和 MHC-II 限制性 CD4+表位。然后，我们对这些保守表位进行了抗原性、变应原性、毒性和可溶性筛选。使用这些表位，我们开发了基于计算机的广谱多表位疫苗，其中包含 Toll 样受体（TLR-4）激动剂。使用 C-IMMSIM 服务器在计算机上评估预测的促炎细胞因子反应。使用 Robetta 和 GalaxyWEB 服务器进行结构建模和细化。最后，通过广泛的 100 纳秒分子动力学模拟评估对接复合物的稳定性，然后再考虑对疫苗进行密码子优化和计算机克隆。

结果

我们在 NS1 和 NS2 蛋白中发现了许多符合这些标准的表位，并开发了基于计算机的广谱疫苗。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-蛋白质对接分析显示了具有与 TLR-4 强结合亲和力的有前途的表位。对接复合物稳定，根均方偏差和根均方波动值最小。最后，计算机克隆的质粒具有高 GC 含量（>0.8 密码子适应指数），表明它们适合在原核系统中表达蛋白疫苗。