布鲁氏菌病,由布鲁氏菌引起的人畜共患疾病,对动物和人类健康都构成重大威胁。在动物中,这种疾病会导致不孕,流产,高烧,而在人类中,症状可能包括反复发烧,疲劳,出汗,肝脾肿大,以及感染后的关节和肌肉疼痛。治疗通常包括长期抗生素治疗,给患者带来巨大的心理和经济负担。虽然接种疫苗对预防至关重要,目前的动物疫苗存在残留毒力等缺点,缺乏安全有效的人类疫苗。因此,布鲁氏菌病疫苗的开发势在必行。在这项研究中,我们利用生物信息学方法设计了一种针对布鲁氏菌的多表位疫苗。靶向血红素转运蛋白BhuA和多糖出口蛋白,我们鉴定了抗原表位,包括六个细胞毒性T淋巴细胞(CTL)优势表位,六个辅助性T淋巴细胞(HTL)优势表位,一个构象B细胞优势表位,和三个线性B细胞优势表位。通过将这些表位与适当的接头连接并掺入Toll样受体(TLR)激动剂(人β-防御素-2)和辅助肽(泛HLA-DR表位),我们构建了多表位疫苗(MEV)。MEV表现出高抗原性,无毒性,非过敏性,非人同源性,稳定性,和溶解度。分子对接分析和分子动力学模拟证实了MEV与受体的相互作用和稳定性(MHCI,MHCII,TLR4)。密码子优化和计算机克隆验证了MEV在大肠杆菌中的翻译效率和成功表达。免疫模拟进一步证明了MEV在诱导稳健免疫应答中的功效。总之,我们的研究结果表明,工程MEV有可能刺激体液和细胞免疫反应,为未来开发安全高效的布鲁氏菌疫苗提供有价值的见解。
Brucellosis, a zoonotic disease caused by Brucella, presents a significant threat to both animal and human health. In animals, the disease can lead to infertility, miscarriage, and high fever, while in humans, symptoms may include recurrent fever, fatigue, sweating, hepatosplenomegaly, and joint and muscle pain following infection. Treatment often involves long-term antibiotic therapy, placing a substantial psychological and financial burden on patients. While vaccination is crucial for prevention, current animal vaccines have drawbacks such as residual virulence, and a safe and effective human vaccine is lacking. Hence, the development of a vaccine for brucellosis is imperative. In this study, we utilized bioinformatics methods to design a multi-epitope vaccine targeting Brucella. Targeting Heme transporter BhuA and polysaccharide export protein, we identified antigenic epitopes, including six cytotoxic T lymphocyte (CTL) dominant epitopes, six helper T lymphocyte (HTL) dominant epitopes, one conformation B cell dominant epitope, and three linear B cell dominant epitopes. By linking these epitopes with appropriate linkers and incorporating a Toll-like receptor (TLR) agonist (human beta-defensin-2) and an auxiliary peptide (Pan HLA-DR epitopes), we constructed the multi-epitope vaccine (MEV). The MEV demonstrated high antigenicity, non-toxicity, non-allergenicity, non-human homology, stability, and solubility. Molecular docking analysis and molecular dynamics simulations confirmed the interaction and stability of the MEV with receptors (MHCI, MHCII, TLR4). Codon optimization and in silico cloning validated the translation efficiency and successful expression of MEV in Escherichia coli. Immunological simulations further demonstrated the efficacy of MEV in inducing robust immune responses. In conclusion, our findings suggest that the engineered MEVs have the potential to stimulate both humoral and cellular immune responses, offering valuable insights for the future development of safe and efficient Brucella vaccines.