School of Biological Sciences, University of the Punjab, Lahore, Pakistan.
King Abdulaziz University, Jeddah, Saudi Arabia.
Sci Rep. 2024 Aug 14;14(1):18883. doi: 10.1038/s41598-024-65593-1.
NS1 is an elusive dengue protein, involved in viral replication, assembly, pathogenesis, and immune evasion. Its levels in blood plasm are positively related to disease severity like thrombocytopenia, hemorrhage, and vascular leakage. Despite its pathogenic roles, NS1 is being used in various vaccine formulations due to its sequence conservancy, ability to produce protective antibodies and low risk for inducing antibody-dependent enhancement. In this study, we have used bioinformatics tools and reported literature to develop an NS1 variant (dNS1). Molecular docking studies were performed to evaluate the receptor-binding ability of the NS1 and dNS1 with TLR4. NS1 and dNS1 (153 to 312 amino acid region) genes were cloned, expressed and protein was purified followed by refolding. Docking studies showed the binding of NS1 and dNS1 with the TLR4 receptor which suggests that N and C-terminal sequences of NS1 are not critical for receptor binding. Antibodies against NS1 and dNS1 were raised in rabbits and binding affinity of anti-dNS1 anti-NS1 sera was evaluated against both NS1 and dNS1. Similar results were observed through western blotting which highlight that N and C-terminal deletion of NS1 does not compromise the immunogenic potential of dNS1 hence, supports its use in future vaccine formulations as a substitute for NS1.
NS1 是一种难以捉摸的登革热蛋白，参与病毒复制、组装、发病机制和免疫逃逸。其在血浆中的水平与血小板减少症、出血和血管渗漏等疾病严重程度呈正相关。尽管 NS1 具有致病性作用，但由于其序列保守性、产生保护性抗体的能力以及低诱导抗体依赖性增强的风险，它被用于各种疫苗制剂中。在这项研究中，我们使用生物信息学工具和报告文献开发了一种 NS1 变体（dNS1）。进行了分子对接研究，以评估 NS1 和 dNS1 与 TLR4 的受体结合能力。克隆、表达并纯化了 NS1 和 dNS1（153 至 312 个氨基酸区域）基因，并进行了复性。对接研究表明 NS1 和 dNS1 与 TLR4 受体结合，这表明 NS1 的 N 和 C 末端序列对于受体结合不是关键的。用 NS1 和 dNS1 抗体制备了兔抗血清，并评估了抗-dNS1 抗 NS1 血清对 NS1 和 dNS1 的结合亲和力。通过 Western blot 观察到了类似的结果，这表明 NS1 的 N 和 C 末端缺失不会影响 dNS1 的免疫原性潜力，因此支持将其用于未来的疫苗制剂中作为 NS1 的替代品。
