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Abstract：
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.
摘要：
NS1是一种难以捉摸的登革热蛋白，参与病毒复制，装配,发病机制，和免疫逃避。其在血浆中的水平与疾病严重程度呈正相关，如血小板减少症，出血,和血管渗漏。尽管其致病作用，NS1由于其序列稳定性而被用于各种疫苗制剂中，产生保护性抗体的能力和诱导抗体依赖性增强的低风险。在这项研究中,我们使用生物信息学工具和报道的文献来开发NS1变异体(dNS1).进行分子对接研究以评估NS1和dNS1与TLR4的受体结合能力。克隆了NS1和dNS1（153至312个氨基酸区域）基因，表达并纯化蛋白质，然后重折叠。对接研究显示NS1和dNS1与TLR4受体的结合,这表明NS1的N和C末端序列对于受体结合不是关键的。在兔中产生抗NS1和dNS1的抗体,并评价抗dNS1抗NS1血清对NS1和dNS1的结合亲和力。通过蛋白质印迹观察到类似的结果，强调NS1的N和C末端缺失不会损害dNS1的免疫原性潜力，因此，支持其在未来的疫苗制剂中用作NS1的替代品。
