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Safety and immunogenicity of an attenuated Chinese pseudorabies variant by dual deletion of TK&gE genes
- Wang, Jichun, Song, Zengcai, Ge, Aimin, Guo, Rongli, Qiao, Yongfeng, Xu, Mengwei, Wang, Zhisheng, Liu, Yamei, Zheng, Yating, Fan, Hongjie, Hou, Jibo
- BMC veterinary research 2018 v.14 no.1 pp. 287
- Aujeszky disease, Suid herpesvirus 1, antibodies, body temperature, death, gene deletion, genes, growth models, herds, homologous recombination, immunogenicity, intramuscular injection, live vaccines, mice, mutagenesis, mutants, nose, piglets, polymerase chain reaction, restriction fragment length polymorphism, signs and symptoms (animals and humans), vaccination, veterinary medicine, viral shedding, virulence, viruses
- BACKGROUND: Since the outbreak of a new emerging virulent pseudorabies virus mutant in Chinese pig herds, intensive research has been focused on the construction of novel gene deletion vaccine based on the variant virulent viruses. An ideal vaccine candidate is expected to have a balanced safety and immunogenicity. RESULTS: From the infectious clone of PRV AH02LA strain, a TK deletion mutant was generated through two-step Red mutagenesis. After homologous recombination with a transfer vector, a TK&gE dual deficient mutant PRV (PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰²) was generated, and its structure verified by PCR, RFLP and sequencing. Growth kinetics test showed that PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰² reached a titer of 10⁷.⁵ TCID₅₀ /mL on ST cells. The PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰² at a dose of 10⁶.⁰ TCID₅₀ /animal was not virulent in mice or 1-day-old piglets with maternal PRV antibodies. No clinical signs or virus shedding were detected in 28~ 35-day-old piglets without maternal PRV antibodies after nasal or intramuscular administration with a dose of 10⁶.⁰ TCID₅₀, although it caused one death of four 1-day-old piglets without maternal PRV antibodies. In the efficiency test of PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰², all four 28~ 35-day-old piglets without PRV antibody in the challenge control showed typical clinical symptoms and virus shedding, and two died at 4~ 5 days post challenge. All piglets in 10⁵.⁰, 10⁴.⁰ and 10³.⁰ TCID₅₀/dose PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰² groups provided complete protection against challenge at only 7 days post intramuscular vaccination. More importantly, PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰² stopped virus shedding in these piglets. In contrast, all four piglets in PRV Bartha K61 vaccine group developed high body temperature (≥40.5 °C) and viral shedding, despite they had mild or even no clinical symptoms. CONCLUSIONS: The constructed TK&gE dual deletion mutant PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰² can reach high titers on ST cells. The live vaccine of PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰² is highly safe, and can not only provide clinical protection but also stops virus shedding. This study suggests that PRVΔᵀᴷ&ᵍᴱ⁻ᴬᴴ⁰² might work as a promising vaccine candidate to combat the PRV variant emerging in Chinese herds since 2011.