Design and preparation of multi-epitope mRMA vaccine for Brucella VirB8, OMO25 and other proteins

Brucellosis is a common zoonotic systemic infectious disease that can cause risks such as miscarriage in animals and can damage various organs in humans to varying degrees. Currently, there are vaccines for brucellosis in animals, but certain risks still exist. Research on vaccines for brucellosis in humans remains somewhat lacking; therefore, we intend to create a safe and effective multi-epitope mRNA vaccine to prevent and protect against the damage caused by brucellosis to humans. By screening the antigenic epitopes of VirB8 from the Brucella IV secretion system, Copper/Zinc Superoxide Dismutase (Cu/Zn-SOD), and Outer Membrane Protein 25 (OMP25), we ultimately identified 4 cytotoxic T lymphocyte (CTL) epitopes, 5 helper T lymphocyte (HTL) epitopes, 3 linear B cell epitopes, and 2 conformational B cell epitopes. All antigenic epitopes exhibit strong antigenicity and pose no potential harm to humans. Subsequently, we linked these epitopes with adjuvants such as cholera toxin B subunit through specific linkers to form the vaccine construct. We then evaluated its physicochemical properties and analyzed its interaction with the TLR4 receptor, dynamically simulating the immune response of the vaccine with the organism, confirming that this structure has the potential to induce a strong humoral and cellular immune response. The results indicate that our designed mRNA construct could become an effective and promising vaccine, providing certain experimental basis for the development of brucellosis vaccines.