PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in mice

Abstract Background The porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause widespread infections in the pig industry worldwide. Currently, multiple PRRSV vaccine candidates are in preclinical or clinical trials, and each has different advantages and limitations. Glycoprot...

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Main Authors: Honghuan Li, Wenxiang Zhang, Wenxing Wang, Yanjie Qiao, Mingguo Xu, Ziwei Liu, Xiaoxiao Gu, Aodi Wu, Zhongchen Ma, Chuangfu Chen, Wei Zhang
Format: Article
Language:English
Published: BMC 2025-05-01
Series:BMC Veterinary Research
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Online Access:https://doi.org/10.1186/s12917-025-04842-5
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author Honghuan Li
Wenxiang Zhang
Wenxing Wang
Yanjie Qiao
Mingguo Xu
Ziwei Liu
Xiaoxiao Gu
Aodi Wu
Zhongchen Ma
Chuangfu Chen
Wei Zhang
author_facet Honghuan Li
Wenxiang Zhang
Wenxing Wang
Yanjie Qiao
Mingguo Xu
Ziwei Liu
Xiaoxiao Gu
Aodi Wu
Zhongchen Ma
Chuangfu Chen
Wei Zhang
author_sort Honghuan Li
collection DOAJ
description Abstract Background The porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause widespread infections in the pig industry worldwide. Currently, multiple PRRSV vaccine candidates are in preclinical or clinical trials, and each has different advantages and limitations. Glycoprotein 4 (GP4) is rich in epitopes, which can induce the body to produce neutralizing antibodies, plays a vital role in causing the host immune response, and is a key target for PRRSV vaccine development. In this study, we developed a novel candidate vaccine immunization strategy combining a subunit vaccine with an adenovirus vector vaccine through prokaryotic and eukaryotic systems expressing GP4. Results In this study, predictive analysis of PRRSV GP4 antigen structures in two expressed modes, and the results showed good antigenicity. The PRRSV GP4 subunit vaccine, as well as the adenovirus vector-based vaccine, were successfully constructed. In the immunization experiment of mouse models, a heterologous primary-boost immunization strategy was implemented: primary immunization with the GP4 subunit vaccine, and boost immunization was followed by an adenovirus vector vaccine. The safety assessment revealed that all candidate vaccine groups demonstrated good safety profiles. With an indirect enzyme-linked immunosorbent assay (ELISA) and neutralizing antibodies, mice in the combined immunization group developed higher levels of PRRSV-specific antibodies with significantly higher neutralizing antibody titers than mice alone. IgG subtype analysis indicated that the proteome favors the Th2-type immune response, while the adenoviral group favors the Th1-type immune response. The secretion levels of cytokines IL-4, IFN-γ, and TNF-α were significantly higher in the serum of the combined immunization group than in the immune group alone. Moreover, the cellular immune response test results showed that the combined immune group significantly enhanced the splenic lymphocyte proliferation capacity, IFN-γ secretion level, and cytokine transcript level. These findings suggest that the heterologous primary-boost immunization strategy of the PRRSV GP4 subunit vaccine developed here, in combination with the adenovirus vaccine, successfully induced strong humoral and cellular immune responses in mice. Conclusions In this study, the PRRSV GP4 subunit and adenovirus vector vaccine were successfully constructed and induced high levels of PRRSV-specific neutralizing antibody and cellular immune responses in mouse models by a heterologous primary-boost immunization strategy. These results support the clinical development of the PRRSV vaccine and bring new hope for PRRSV prevention and control strategies in the swine industry.
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spelling doaj-art-60759c2c07ab428aba6b6db1f65c72492025-08-20T01:59:56ZengBMCBMC Veterinary Research1746-61482025-05-0121111410.1186/s12917-025-04842-5PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in miceHonghuan Li0Wenxiang Zhang1Wenxing Wang2Yanjie Qiao3Mingguo Xu4Ziwei Liu5Xiaoxiao Gu6Aodi Wu7Zhongchen Ma8Chuangfu Chen9Wei Zhang10College of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Animal Science and Technology, Shihezi UniversityCollege of Veterinary Medicine, Xinjiang Agricultural UniversityAbstract Background The porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause widespread infections in the pig industry worldwide. Currently, multiple PRRSV vaccine candidates are in preclinical or clinical trials, and each has different advantages and limitations. Glycoprotein 4 (GP4) is rich in epitopes, which can induce the body to produce neutralizing antibodies, plays a vital role in causing the host immune response, and is a key target for PRRSV vaccine development. In this study, we developed a novel candidate vaccine immunization strategy combining a subunit vaccine with an adenovirus vector vaccine through prokaryotic and eukaryotic systems expressing GP4. Results In this study, predictive analysis of PRRSV GP4 antigen structures in two expressed modes, and the results showed good antigenicity. The PRRSV GP4 subunit vaccine, as well as the adenovirus vector-based vaccine, were successfully constructed. In the immunization experiment of mouse models, a heterologous primary-boost immunization strategy was implemented: primary immunization with the GP4 subunit vaccine, and boost immunization was followed by an adenovirus vector vaccine. The safety assessment revealed that all candidate vaccine groups demonstrated good safety profiles. With an indirect enzyme-linked immunosorbent assay (ELISA) and neutralizing antibodies, mice in the combined immunization group developed higher levels of PRRSV-specific antibodies with significantly higher neutralizing antibody titers than mice alone. IgG subtype analysis indicated that the proteome favors the Th2-type immune response, while the adenoviral group favors the Th1-type immune response. The secretion levels of cytokines IL-4, IFN-γ, and TNF-α were significantly higher in the serum of the combined immunization group than in the immune group alone. Moreover, the cellular immune response test results showed that the combined immune group significantly enhanced the splenic lymphocyte proliferation capacity, IFN-γ secretion level, and cytokine transcript level. These findings suggest that the heterologous primary-boost immunization strategy of the PRRSV GP4 subunit vaccine developed here, in combination with the adenovirus vaccine, successfully induced strong humoral and cellular immune responses in mice. Conclusions In this study, the PRRSV GP4 subunit and adenovirus vector vaccine were successfully constructed and induced high levels of PRRSV-specific neutralizing antibody and cellular immune responses in mouse models by a heterologous primary-boost immunization strategy. These results support the clinical development of the PRRSV vaccine and bring new hope for PRRSV prevention and control strategies in the swine industry.https://doi.org/10.1186/s12917-025-04842-5PRRSVGlycoprotein 4Subunit vaccineAdenovirusHeterologous prime-boost immunization strategyVaccine
spellingShingle Honghuan Li
Wenxiang Zhang
Wenxing Wang
Yanjie Qiao
Mingguo Xu
Ziwei Liu
Xiaoxiao Gu
Aodi Wu
Zhongchen Ma
Chuangfu Chen
Wei Zhang
PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in mice
BMC Veterinary Research
PRRSV
Glycoprotein 4
Subunit vaccine
Adenovirus
Heterologous prime-boost immunization strategy
Vaccine
title PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in mice
title_full PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in mice
title_fullStr PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in mice
title_full_unstemmed PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in mice
title_short PRRSV GP4 subunit vaccine combined with adenovirus heterologous prime-boost immunization strategy induced a significant immune response in mice
title_sort prrsv gp4 subunit vaccine combined with adenovirus heterologous prime boost immunization strategy induced a significant immune response in mice
topic PRRSV
Glycoprotein 4
Subunit vaccine
Adenovirus
Heterologous prime-boost immunization strategy
Vaccine
url https://doi.org/10.1186/s12917-025-04842-5
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