Department of Animal Science, Faculty of Agriculture, Ilam University, Ilam, Iran.
Department of Laboratory and Clinical Sciences, Faculty of Para-Veterinary, Ilam University, Ilam, Iran.
J Biomol Struct Dyn. 2023;41(24):15460-15484. doi: 10.1080/07391102.2023.2188962. Epub 2023 Mar 16.
Brucellosis is a zoonotic caused by the which is a well-known infectious disease agent in domestic animals and if transmitted, it can cause infection in humans. Because brucellosis is contagious, its control depends on the eradication of the animal disease in farms. There are two vaccines based on the killed and/or weakened bacteria against and , but no recombinant vaccine is available for preventing the disease. The present study was designed to develop a multi-epitope vaccine against of and using virB10, Omp31 and Omp16 antigens by the prediction of T lymphocytes, T cell cytotoxicity and IFN-γ epitopes. 50S L7/L12 Ribosomal protein from Mycobacterium tuberculosis was used as a bovine TLR4 and TLR9 agonist. GPGPG, AAY and KK linkers were used as a linker. construct was well-integrated in the pET-32a Shuttle vector with I and III restriction enzymes. The final construct contained 769 amino acids, that it was soluble protein of about ∼82 kDa after expression in the SHuffle host. Modeled protein analysis based on the tertiary structure validation, molecular docking studies, molecular dynamics simulations results like RMSD, Gyration and RMSF as well as MM/PBSA analysis showed that this protein has a stable construct and is capable being in interaction with bovine TLR4 and TLR9. Analysis of the data obtained suggests that the proposed vaccine can induce the immune response by stimulating T- and B-cells, and may be used for prevention and remedial purposes, against and Communicated by Ramaswamy H. Sarma.
布鲁氏菌病是一种由 引起的人畜共患病，已知其是家畜中的一种传染性病原体，如果传播，它会导致人类感染。由于布鲁氏菌病具有传染性，其控制取决于农场动物疾病的根除。有两种基于灭活和/或减毒细菌的疫苗针对 和 ，但目前尚无预防该病的重组疫苗。本研究旨在通过预测 T 淋巴细胞、T 细胞细胞毒性和 IFN-γ 表位，使用 virB10、Omp31 和 Omp16 抗原开发针对 和 的多表位疫苗。结核分枝杆菌 50S L7/L12 核糖体蛋白被用作牛 TLR4 和 TLR9 激动剂。GPGPG、AAY 和 KK 接头用作接头。 构建体与 I 和 III 限制酶一起很好地整合到 pET-32a 穿梭载体中。最终构建体包含 769 个氨基酸，在 SHuffle 宿主中表达后，它是约 ∼82 kDa 的可溶性蛋白。基于三级结构验证、分子对接研究、分子动力学模拟结果（如 RMSD、Gyration 和 RMSF 以及 MM/PBSA 分析）的建模蛋白分析表明，该蛋白具有稳定的结构，能够与牛 TLR4 和 TLR9 相互作用。对获得的数据的分析表明，所提出的疫苗可以通过刺激 T 细胞和 B 细胞来诱导免疫反应，并可能用于预防和治疗目的，针对 和 。通讯作者为 Ramaswamy H. Sarma。
