Eukaryotic expression of GP5 and M protein of porcine reproductive and respiratory syndrome virus and immunogenicity evaluation.

Huicong Lou; Runshan Lin; Yabo LI; Yuna Zhao; Pengtao Jiao; Tingrong Luo; Wenjun Liu

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Eukaryotic expression of GP5 and M protein of porcine reproductive and respiratory syndrome virus and immunogenicity evaluation.

Author: Huicong LOU ¹ ; Runshan LIN ² ; Yabo LI ¹ ; Yuna ZHAO ¹ ; Pengtao JIAO ² ; Tingrong LUO ¹ ; Wenjun LIU ¹
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1. Animal Science and Technology College, Guangxi University, Nanning 530004, Guangxi, China.
2. Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Publication Type:Journal Article
Keywords: GP5 recombinant protein; M recombinant protein; genetic evolution analysis; immunogenicity evaluation; porcine reproductive and respiratory syndrome virus (PRRS)
MeSH: Humans; Animals; Swine; Porcine respiratory and reproductive syndrome virus/genetics*; Porcine Reproductive and Respiratory Syndrome/prevention & control*; HEK293 Cells; Viral Envelope Proteins/genetics*; Antibodies, Viral; Viral Vaccines/genetics*; Recombinant Proteins; Vaccines, Subunit
From: Chinese Journal of Biotechnology 2023;39(12):4809-4823
CountryChina
Language:Chinese
Abstract: In order to understand the prevalence and evolution of porcine reproductive and respiratory syndrome virus (PRRSV) in China and to develop subunit vaccine against the epidemic lineage, the genetic evolution analysis of PRRSV strains isolated in China from 2001 to 2021 was performed. The representative strains of the dominant epidemic lineage were selected to optimize the membrane protein GP5 and M nucleotide sequences, which were used, with the interferon and the Fc region of immunoglobulin, to construct the eukaryotic expression plasmids pCDNA3.4-IFNα-GP5-Fc and pCDNA3.4-IFNα-M-Fc. Subsequently, the recombinant proteins IFNα-GP5-Fc and IFNα-M-Fc were expressed by HEK293T eukaryotic expression system. The two recombinant proteins were mixed with ISA206VG adjuvant to immunize weaned piglets. The humoral immunity level was evaluated by ELISA and neutralization test, and the cellular immunity level was detected by ELISPOT test. The results showed that the NADC30-like lineage was the main epidemic lineage in China in recent years, and the combination of IFNα-GP5-Fc and IFNα-M-Fc could induce high levels of antibody and cellular immunity in piglets. This study may facilitate the preparation of a safer and more effective new PRRSV subunit vaccine.