A Pool of Bacterium-like Particles Displaying African Swine Fever Virus Antigens Induces Both Humoral and Cellular Immune Responses in Pigs

Background/Objectives: African swine fever (ASF), caused by African swine fever virus (ASFV), poses a significant threat to the global swine industry. This underscores the urgent need for safe and effective ASF vaccines. Methods: Here, we constructed five bacterium-like particles (BLPs) that each display one of the five ASFV antigens (F317L, H171R, D117L, B602L, and p54) based on the Gram-positive enhancer matrix-protein anchor (GEM-PA) system. GEM is a bacterial particle that contains only peptidoglycan, while PA is composed of three lysin motifs (Lysm) derived from the C-terminus of the AcmA protein, capable of non-covalently binding to GEM. By fusing the ASFV antigens with PA, the ASFV antigens can be firmly attached to the surface of GEM. Subsequently, the piglets were immunized via intramuscular injection with a mixture of BLPs-F317L, BLPs-H171R, BLPs-D117L, BLPs-B602L, and BLPs-p54. Results: The results showed that the piglets developed detectable serum IgG antibodies 2 weeks after the first immunization, and these high antibody levels were maintained 4 weeks after the booster immunization. Moreover, these piglets produced more IFN-gamma-producing lymphocytes than the control piglets. Conclusions: The data indicate that the generated BLPs mixture can stimulate both humoral and cellular immune responses in piglets, these five ASFV proteins are promising antigens, and the BLPs generated represent candidate ASF vaccines.