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Immunogenicity evaluation of a recombinant pseudorabies virus co-expressing PCV2 and PCV3 capsid proteins in mice and piglets
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Immunogenicity evaluation of a recombinant pseudorabies virus co-expressing PCV2 and PCV3 capsid proteins in mice and piglets
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Immunogenicity evaluation of a recombinant pseudorabies virus co-expressing PCV2 and PCV3 capsid proteins in mice and piglets
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Journal Article
Immunogenicity evaluation of a recombinant pseudorabies virus co-expressing PCV2 and PCV3 capsid proteins in mice and piglets
2025
Overview
Porcine circovirus type 2 (PCV2), porcine circovirus type 3 (PCV3), and pseudorabies virus (PRV) are major pathogens posing significant threats to the swine industry. Viral evolution and mutations have limited the efficacy of current commercial vaccines, necessitating the development of more effective prophylactic strategies. In this study, a novel recombinant virus strain, designated as rPRV-ΔTK-PCV3(Cap)/ΔgIgE-PCV2(Cap), was generated using PRV SX-10 variant as the backbone. CRISPR/Cas9-mediated deletion of TK and gE/gI genes was performed, followed by insertion of PCV3 and PCV2 capsid protein genes into the respective loci. The engineered recombinant strain demonstrated stable proliferation in BHK-21 cells, efficiently expressed heterologous PCV3 and PCV2 capsid proteins, while maintaining biological properties comparable to its parental strain. The rPRV-ΔTK-PCV3(Cap)/ΔgIgE-PCV2(Cap) demonstrated favorable safety and immunogenicity profiles in mice and piglets, eliciting robust immune responses characterized by high titers of specific antibodies against PRV, PCV3, and PCV2, along with significantly elevated levels of cytokines (IFN-γ, IL-2, and IL-4). Histopathological analysis and viral load quantification demonstrated that rPRV-ΔTK-PCV3(Cap)/ΔgIgE-PCV2(Cap) immunization significantly attenuated tissue lesions and decreased viral copies of PRV and PCV in mice and piglets. Collectively, these findings suggest that rPRV-ΔTK-PCV3(Cap)/ΔgIgE-PCV2(Cap) serves as a promising candidate vaccine against PRV and PCV infections.
•Generated novel recombinant virus rPRV-ΔTK-PCV3(Cap)/ΔgIgE-PCV2(Cap) via CRISPR/Cas9.•Demonstrated stable virus proliferation and efficient PCV3/PCV2 capsid protein expression.•Induced robust immune responses against PRV, PCV3, and PCV2.•Provided valuable insights for the development of trivalent vaccine candidates.
Publisher
Elsevier Ltd,Elsevier Limited
Subject
/ Animals
/ Cap
/ Capsid Proteins - immunology
/ Circoviridae Infections - immunology
/ Circoviridae Infections - prevention & control
/ Circoviridae Infections - veterinary
/ CRISPR
/ Enzymes
/ Female
/ Genes
/ Genomes
/ Herpesvirus 1, Suid - genetics
/ Herpesvirus 1, Suid - immunology
/ Hogs
/ Kinases
/ Mice
/ PCV2
/ PCV3
/ Plasmids
/ Proteins
/ Pseudorabies - prevention & control
/ Pseudorabies Vaccines - administration & dosage
/ Pseudorabies Vaccines - immunology
/ Swine
/ Swine Diseases - prevention & control
/ Vaccines
/ Vaccines, Synthetic - administration & dosage
/ Vaccines, Synthetic - genetics
/ Vaccines, Synthetic - immunology
/ Viral Vaccines - administration & dosage
/ Viruses
