Feline panleukopenia, caused by feline parvovirus (FPV), is a highly infectious disease characterized by leucopenia and hemorrhagic gastroenteritis that severely affects the health of large wild Felidae. In this study, tiger FPV virus-like particles (VLPs) were developed using the baculovirus expression system. The VP2 gene from an infected Siberian tiger (Panthera tigris altaica) was used as the target gene. The key amino acids of this gene were the same as those of FPV, whereas the 101st amino acid was the same as that of canine parvovirus. Indirect immunofluorescence assay (IFA) results demonstrated that the VP2 protein was successfully expressed. SDS-PAGE and Western blotting (WB) results showed that the target protein band was present at approximately 65 kDa. Electron micrograph analyses indicated that the tiger FPV VLPs were successfully assembled and were morphologically similar to natural parvovirus particles. The hemagglutination (HA) titer of the tiger FPV VLPs was as high as 1:2¹⁸. The necropsy and tissue sections at the cat injection site suggested that the tiger FPV VLPs vaccine was safe. Antibody production was induced in cats after subcutaneous immunization, with a >1:2¹⁰ hemagglutination inhibition (HI) titer that persisted for at least 12 months. These results demonstrate that tiger FPV VLPs might provide a vaccine to prevent FPV-associated disease in the tiger.

Sudan virus (SUDV) causes severe lethal hemorrhagic fever in humans and nonhuman primates. The most effective and economical way to protect against Sudan ebolavirus disease is prophylactic vaccination. However, there are no licensed vaccines to prevent SUDV infections. In this study, a bacterium-like particle (BLP)-based vaccine displaying the extracellular domain of the SUDV glycoprotein (eGP) was developed based on a gram-positive enhancer matrix-protein anchor (GEM-PA) surface display system. Expression of the recombinant GEM-displayed eGP (eGP-PA-GEM) was verified by Western blotting and immunofluorescence assays. The SUDV BLPs (SBLPs), which were mixed with Montanide ISA 201VG plus Poly (I:C) combined adjuvant, could induce high SUDV GP-specific IgG titers of up to 1:40,960 and robust virus-neutralizing antibody titers reached 1:460. The SBLP also elicited T-helper 1 (Th1) and T-helper 2 (Th2) cell-mediated immunity. These data indicate that the SBLP subunit vaccine has the potential to be developed into a promising candidate vaccine against SUDV infections.

The Crimean Congo Hemorrhagic Fever Virus (CCHFV) is a tick-borne bunyavirus of the Narovirus genus, which is the causative agent of Crimean Congo Hemorrhagic Fever (CCHF). CCHF is endemic in Africa, the Middle East, Eastern Europe and Asia, with a high case-fatality rate of up to 50% in humans. Currently, there are no approved vaccines or effective therapies available for CCHF. The GEM-PA is a safe, versatile and effective carrier system, which offers a cost-efficient, high-throughput platform for recovery and purification of subunit proteins for vaccines. In the present study, based on a GEM-PA surface display system, a GEM-PA based vaccine expressing three subunit vaccine candidates (G-GP, including G-eG_N, G-eG_C and G-NAb) of CCHFV was developed, displaying the ectodomains of the structural glycoproteins eG_N, eG_C and NAb, respectively. According to the immunological assays including indirect-ELISA, a micro-neutralization test of pseudo-virus and ELISpot, 5 μg GPBLP₃ combined with Montanide ISA 201VG plus Poly (I:C) adjuvant (A-G-GP-5 μg) elicited GP-specific humoral and cellular immunity in BALB/c mice after three vaccinations via subcutaneous injection (s.c.). The consistent data between IgG subtype and cytokine detection, ELISpot and cytokine detection indicated balanced Th1 and Th2 responses, of which G-eG_N vaccines could elicit a stronger T-cell response post-vaccination, respectively. Moreover, all three vaccine candidates elicited high TNF-α, IL-6, and IL-10 cytokine levels in the supernatant of stimulated splenocytes in vitro. However, the neutralizing antibody (nAb) was only detected in A-G-eG_C and A-G-eG_C vaccination groups with the highest neutralizing titer of 128, suggesting that G-eG_C could elicit a stronger humoral immune response. In conclusion, the GEM-PA surface display system could provide an efficient and convenient purification method for CCHFV subunit antigens, and the G-GP subunit vaccine candidates will be promising against CCHFV infections with excellent immunogenicity.

Introduction: Porcine reproductive and respiratory syndrome virus (PRRSV) was first reported in pigs in China in 1996. At present, multiple PRRSV lineages and sub-lineages are circulating in Chinese swine herds. Methods: In this study, PRRSV ORF5 sequences collected from 2019 to 2024 were analyzed to investigate their spatiotemporal distribution, molecular evolution, and major amino acid mutations. Results: A total of 674 sequences were obtained from 25 provinces between 2019 and 2024, and their geographical distribution was generally consistent with the major pig-producing regions in China. Phylogenetic analysis revealed that these PRRSV ORF5 sequences mainly belonged to lineage 1 (L1A, L1C), L3, L5 (L5A), and L8 (L8E). From 2019 to 2024, the proportion of L1C initially increased, followed by a decrease and a subsequent rebound, while L1C remained the predominant sub lineage. The proportion of L1A showed a relative increase, whereas L3 was detected at very low levels. Meanwhile, L5A and L8E exhibited a downward trend. Amino acid mutations were observed in the signal peptide, neutralizing epitopes, and T-cell epitopes among different sub-lineages. Some mutations were variable, while others were sub-lineage-specific. Notably, partial L1C strains exhibited an amino acid deletion at position 34, which may affect the viral immune evasion mechanisms. Nucleotide and amino acid identities initially decreased and then increased, with the most pronounced divergence observed in 2021. Discussion: Continuous monitoring of the molecular epidemiology of PRRSV is essential and will provide a scientific basis for the further prevention and control of PRRSV in China.