Faculty of Biotechnology and Genetic Engineering, Sylhet Agricultural University, Sylhet 3100, Bangladesh; Department of Microbial Biotechnology, Sylhet Agricultural University, Sylhet 3100, Bangladesh.
Faculty of Biotechnology and Genetic Engineering, Sylhet Agricultural University, Sylhet 3100, Bangladesh; Department of Pharmaceuticals and Industrial Biotechnology, Sylhet Agricultural University, Sylhet 3100, Bangladesh.
Infect Genet Evol. 2019 Oct;74:103936. doi: 10.1016/j.meegid.2019.103936. Epub 2019 Jun 21.
Norovirus is known as a major cause of several acute gastroenteritis (AGE) outbreaks each year. A study was conducted to develop a unique multi epitope subunit vaccine against human norovirus by adopting reverse vaccinology approach. The entire viral proteome of Norwalk virus was retrieved and allowed for further in silico study to predict highly antigenic epitopes through antigenicity, transmembrane topology screening, allergenicity assessment, toxicity analysis, population coverage analysis and molecular docking approach. Capsid protein VP1 and protein VP2 were identified as most antigenic viral proteins which generated a plethora of antigenic epitopes. Physicochemical properties and secondary structure of the designed vaccine were assessed to ensure its thermostability, hydrophilicity, theoretical PI and structural behavior. Molecular docking analysis of the refined vaccine with different MHCs and human immune TLR8 receptor demonstrated higher binding interaction as well. Complexed structure of the modeled vaccine and TLR8 showed minimal deformability at molecular level. The designed construct was reverse transcribed and adapted for E. coli strain K12 prior to insertion within pET28a(+) vector for its heterologous cloning and expression, and sequence of vaccine constructs showed no similarity with human proteins. However, the study could initiate in vitro and in vivo studies regarding effective vaccine development against human norovirus.
诺如病毒是导致每年数次急性肠胃炎（AGE）爆发的主要原因之一。本研究采用反向疫苗学方法，旨在开发针对人类诺如病毒的独特多表位亚单位疫苗。从诺如病毒中获取了整个病毒蛋白组，并进行了进一步的计算机研究，通过抗原性、跨膜拓扑筛选、变应原性评估、毒性分析、人群覆盖率分析和分子对接方法预测高抗原性表位。衣壳蛋白 VP1 和蛋白 VP2 被鉴定为最具抗原性的病毒蛋白，产生了大量的抗原表位。对设计疫苗的物理化学性质和二级结构进行了评估，以确保其热稳定性、亲水性、理论 PI 和结构行为。对不同 MHC 和人类免疫 TLR8 受体的精制疫苗进行分子对接分析表明，其结合相互作用更高。建模疫苗和 TLR8 的复合物结构在分子水平上显示出最小的可变形性。设计的构建体被反转录，并适应于大肠杆菌菌株 K12，然后插入 pET28a(+)载体中进行异源克隆和表达，疫苗构建体的序列与人蛋白没有相似性。然而，该研究可以启动针对人类诺如病毒的有效疫苗开发的体外和体内研究。
