■大肠杆菌是引起动物领域各种感染的严重病原体之一,比如新生儿小牛腹泻,这是在生命的第一天与腹泻相关的死亡率的原因。
■当前的工作旨在基于肠产毒性大肠杆菌(ETEC)的菌毛蛋白K99和免疫信息学设计针对小牛大肠杆菌感染的有效且安全的多表位疫苗候选物。
■产生了K99蛋白的保守序列,然后是高度抗原性的,不过敏,和重叠表位用于构建多表位疫苗。五THL,六个MHCII,和四个β细胞表位被靶向以创建候选物。候选疫苗是利用15个表位和三种类型的接头产生的,两种类型的非翻译区(UTR)人血红蛋白亚基β(HBB),UTRβ-珠蛋白(Rabb),和RpfE蛋白作为免疫调节佐剂。
■对构建的蛋白进行免疫信息学分析表明,该蛋白具有抗原性(抗原性评分为0.8841),稳定,非过敏原,和可溶性。此外,构建的蛋白质的免疫信息学和理化分析显示,不过敏,可溶性,亲水性,和酸性PI(等电点)。9.34。进行候选疫苗与toll样受体TLR3的对接,结果显示免疫受体和疫苗之间有很强的相互作用。最后,构建体在大肠杆菌中的表达效率是通过将疫苗序列计算克隆到Pet28a中来估计的。
■免疫信息学和计算机模拟方法的结果表明,设计的疫苗是抗原性的,稳定,能够与免疫细胞受体结合。我们的结果解释了所提出的多表位mRNA疫苗是针对小牛大肠杆菌感染的良好预防选择。
UNASSIGNED: Escherichia coli is one of the serious pathogens causing various infections in the animal field, such as neonatal calf diarrhea, which is responsible for mortality associated with diarrhea during the first days of life.
UNASSIGNED: Current work is aimed at designing an effective and safe multiepitope vaccine candidate against E. coli infection in calves based on the fimbrial protein K99 of Enterotoxigenic E. coli (ETEC) and Immuno-informatics.
UNASSIGNED: A conserved sequence of K99 protein was generated, and then highly antigenic, nonallergic, and overlapped epitopes were used to construct a multiepitope vaccine. Five THL, six MHC II, and four beta cell epitopes were targeted to create the candidate. The candidate vaccine was produced utilizing 15 epitopes and three types of linkers, two types of untranslated region (UTR) human hemoglobin subunit beta (HBB), UTR beta-globin (Rabb), and RpfE protein as an immunomodulation adjuvant.
UNASSIGNED: Immuno-informatics analysis of the constructed protein showed that the protein was antigenic (antigenic score of 0.8841), stable, nonallergen, and soluble. Furthermore, the Immuno-informatics and physiochemical analysis of the constructed protein showed a stable, nonallergic, soluble, hydrophilic, and acidic PI (isoelectric point). of 9.34. Docking of the candidate vaccine with the toll-like receptor TLR3 was performed, and results showed a strong interaction between the immune receptor and the vaccine. Finally, the expression efficiency of the construct in E. coli was estimated via computational cloning of the vaccine sequence into Pet28a.
UNASSIGNED: Results of immunoinformatics and in silico approaches reveal that the designed vaccine is antigenic, stable, and able to bind to the immune cell receptors. Our results interpret the proposed multiepitope mRNA vaccine as a good preventive option against E. coli infection in calves.