Selection, Design and Immunogenicity Studies of ASFV Antigens for Subunit mRNA Cocktail Vaccines with Specific Immune Response Profiles

Development of safe and effective subunit vaccines for controlling African Swine Fever Virus (ASFV) infection has been hampered by a lack of protective viral antigens, complex virion structures, and multiple mechanisms of infection. Here, we selected ASFV antigens based on their localization on the virion, known functions, and homologies to the subunits of the protective vaccinia virus vaccine. We also engineered viral capsid proteins for inducing optimal antibody responses and designed T cell-directed antigen for inducing broad and robust cellular immunity. The selected antigens in lipid nanoparticle-mRNA formulations were evaluated for immunogenicity in both mice and pigs with concordant results. Different antigens induced divergent immune response profiles, including the levels of IgG and T cell responses and effector functions of anti-sera. We further developed a computational approach to combine antigens into cocktails for inducing specific immune response profiles and validated candidate cocktail vaccines in mice. Our results provide a basis for further evaluating candidate subunit mRNA vaccines in challenge studies. Teaser Novel strategies to develop subunit vaccines for ASFV and other complex large DNA viruses.