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| Expression and Immunogenicity Analysis of PRRSV GP5 Antigenic Peptide Recombinant Adenovirus |
| LI Hong-Huan1, LIU Qing-Qing1, QIAO Yan-Jie1, WANG Wen-Xing1, HOU Hui-Lin1, ZHANG Yan-Bing1, WANG Zhen1,2,*, CHEN Chuang-Fu1,2,* |
1 College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2 Ministry of Education Collaborative Innovation Center for Healthy Sheep Breeding and Zoonotic Disease Prevention and Control, Shihezi 832003, China |
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Abstract Porcine reproductive and respiratory syndrome (PRRS) has shown a persistent epidemic trend in our country, with inactivated vaccines and attenuated live vaccines being the primary options domestically. However, these vaccines still have certain limitations. This study aims to develop a safe and efficient PRRS vaccine. By predicting the concentrated region of antigenic epitopes of glycoprotein 5 (GP5) through bioinformatics, a GP5 antigenic peptide recombinant Adenovirus was constructed. After immunizing mice (Mus musculus), its safety, neutralizing antibody titer, lymphocyte interferon-gamma (IFN-γ) secretion level, lymphocyte proliferation, and cytokine transcription level were evaluated to analyze its immunogenic effect. The results showed that PCR and enzyme digestion successfully constructed the recombinant adenoviral plasmid pAdTrack-GP5aa130-170. Western blot identification of the packaged Adenovirus confirmed the construction of a recombinant Adenovirus capable of stably expressing the target protein. After immunization with recombinant Adenovirus, the mice showed good weight gain and mental state. The neutralizing antibody titer, lymphocyte IFN-γ release level, and lymphocyte proliferation rate were significantly higher than the control group (P<0.05), with no significant difference compared to the commercial vaccine. The transcription levels of lymphocyte interleukin 4 (IL-4), IL-6, and IL-10 were extremely significantly higher than that in the control and commercial vaccine groups (P<0.01). At the same time, there was no significant difference in tumor necrosis factor-α (TNF-α) expression level. The secretion levels of IL-4, IFN-γ, and TNF-α in the serum were extremely significantly higher than the control group (P<0.01). These results indicated that this study successfully constructed a recombinant Adenovirus expressing the GP5 antigenic epitope region, which could induce a high level of neutralizing antibodies and cellular immune responses in mice after immunization, and had good immunogenicity. The findings provide a reference for developing a novel vaccine against PRRS.
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Received: 11 November 2024
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Corresponding Authors:
*wzhen2018@shzu.edu.cn; ccf-xb@163.com
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