Immunogenic evaluation of pseudorabies virus gB protein expressed in the baculovirus-insect cell system.

Jin WANG; Kai WANG; Ying ZHANG; Shuzhen TAN; Shiqi SUN; Huichen GUO; Shuanghui YIN; Jiaqiang NIU

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Immunogenic evaluation of pseudorabies virus gB protein expressed in the baculovirus-insect cell system.

Author: Jin WANG ¹ ; Kai WANG ² ; Ying ZHANG ² ; Shuzhen TAN ² ; Shiqi SUN ² ; Huichen GUO ² ; Shuanghui YIN ² ; Jiaqiang NIU ¹
Author Information

1. Key Laboratory of Hydatid Disease Control in Tibet, Ministry of Agriculture and Rural Affairs/Key Laboratory of Universities of Tibet Plateau Animal Disease Research, College of Animal Science, Xizang Agricultural and Animal Husbandry University, Nyingchi 860000, Xizang, China.
2. State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine and Biosafety, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, Gansu, China.
Publication Type:Journal Article
Keywords: gB protein; immunogenic evaluation; insect cell expression; mice; pseudorabies virus
MeSH: Animals; Mice; Baculoviridae/metabolism*; Viral Envelope Proteins/biosynthesis*; Herpesvirus 1, Suid/genetics*; Pseudorabies/immunology*; Swine; Pseudorabies Vaccines/genetics*; Antibodies, Viral/blood*; Insecta/cytology*; Mice, Inbred BALB C; Female; Viral Vaccines/immunology*
From: Chinese Journal of Biotechnology 2025;41(7):2694-2706
CountryChina
Language:Chinese
Abstract: Pseudorabies (PR) is an infectious disease caused by the pseudorabies virus (PRV), affecting various domesticated and wild animals. Since pigs are the only natural hosts of PRV, PR poses a serious threat to the pig farming industry. Currently, PR is primarily prevented through vaccination with inactivated vaccines or genetically modified attenuated live vaccines. Developing safe and effective genetically engineered vaccines would facilitate the eradication and control of PR. In this study, the PRV vaccine strain Bartha-K61 was used as the reference strain. The gB protein was expressed via the baculovirus-insect cell expression system. Non-denaturing gel electrophoresis confirmed that the gB protein could form a trimeric structure. The purified gB protein was used to immunize mice, and the immune effect was evaluated by a challenge test. The results showed that the gB antigen induced a strong immune response in mice, with the serum-neutralizing antibody titer above 1:70. The lymphocyte stimulation index reached more than 1.29, and the level of (interferon gamma, IFN-γ) release was higher than 100 pg/mL. After immunization, mice were challenged with the virus at a dose of 10⁴ TCID₅₀/mL, 200 μL per mouse, and the clinical protection rate was 100%. Immunohistochemistry, histopathological section, and tissue viral load results showed that the pathological damage and viral load in the gB-immunized group were significantly lower than those in the PBS group. In summary, the gB protein obtained in this study induced strong humoral and cellular immune responses in mice, laying a foundation for developing a recombinant gB protein subunit vaccine.