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Construction of Two Recombinant Pseudorabies Viruses with Deletion of Virulence Genes and Evaluation of Their Immune Protection in Mice and Piglets

文献类型： 外文期刊

作者： Wang, Shanghui 1 ; Han, Longfei 1 ; Yu, Jimin 1 ; Ye, Guangqiang 1 ; Liu, Hongyang 1 ; Liu, Yunfei 1 ; Zhou, Qiongqiong 1 ; Zhang, Zhaoxia 1 ; Weng, Changjiang 1 ;

作者机构： 1.Chinese Acad Agr Sci CAAS, Harbin Vet Res Inst, Div Fundamental Immunol, State Key Lab Anim Dis Control & Prevent, Harbin 150069, Peoples R China

2.Zhejiang A&F Univ, Coll Anim Sci & Technol, Hangzhou 311300, Peoples R China

3.Zhejiang A&F Univ, Coll Vet Med, Hangzhou 311300, Peoples R China

4.Heilongjiang Prov Key Lab Vet Immunol, Harbin 150069, Peoples R China

关键词： pseudorabies virus (PRV); live attenuated vaccine; CRISPR-Cas9

期刊名称：VACCINES （ 影响因子：3.4； 五年影响因子：3.7 ）

ISSN：

年卷期： 2025 年 13 卷 4 期

页码：

收录情况： SCI

摘要： Background: Since 2011, re-emerging pseudorabies virus (PRV) variant strains have been widespread in swine herds immunized with the classical PRV vaccine in China, suggesting that it is necessary to develop a new vaccine against these PRV variant strains. Methods: Here, based on a PRV mutant strain isolated in Jinmen (JM), two recombinant strains were constructed using CRISPR/Cas9 technology, including PRV-JM-Delta EK with the deletion of the gE and TK genes and PRV-JM-Delta EI92K with the deletion of the gE, gI, US2, US9, and TK genes. Results: A one-step growth curve and plaque assay revealed that the cell-to-cell transmission ability of PRV-JM-Delta EI92K was lower than that of PRV-JM-Delta EK. However, the replication ability of PRV-JM-Delta EI92K was approximately 10 times higher than that of PRV-JM-Delta EK, similar to wild-type PRV-JM. The intramuscular injection of 106 TCID50 of PRV-JM-Delta EK or PRV-JM-Delta EI92K could not cause death in mice, and both could produce specific antibodies against gB and gD. The survival rate of mice immunized with both recombinant viruses was 100% when the mice were challenged by the PRV-JM strain. Histopathological sections from the PRV-JM-Delta EK group showed milder pathological changes compared to the PRV-JM-Delta EI92K group, proving that PRV-JM-Delta EK provided more effective protection. In pigs injected with 106 TCID50 of PRV-JM-Delta EK or PRV-JM-Delta EI92K, their body temperature did not rise, and their weight gain was not affected. Both recombinant viruses could induce the production of gB- and gD-specific antibodies and neutralizing antibodies. After the challenge of the PRV-JM virus, neutralizing antibody production was rapidly induced and lasted for at least 3 weeks. Pigs immunized with both PRV-JM-Delta EI92K and PRV-JM-Delta EK had a 100% survival rate, demonstrating that both recombinant viruses could provide effective protection. Conclusions: Compared with PRV-JM-Delta EK, PRV-JM-Delta EI92K had better safety. In conclusion, we constructed two PRV recombinant viruses, which have the potential to be used as a live carrier vaccine.

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