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Construction of pseudorabies virus variant attenuated vaccine: codon deoptimization of US3 and UL56 genes based on PRV gE/TK deletion strain

文献类型： 外文期刊

作者： Xu, Mengwei 1 ; Zhu, Laixu 1 ; Ge, Aimin 5 ; Liu, Yamei 1 ; Chen, Saisai 1 ; Wei, Ziwen 1 ; Zheng, Yating 1 ; Tong, Ling 1 ; Wang, Zhisheng 1 ; Fei, Rongmei 4 ; Wang, Jichun 1 ; Zhang, Chuanjian 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Immunol & Engn, Natl Res Ctr Engn & Technol Vet Biol, Jiangsu Key Lab Food Qual & Safety,State Key Lab C, Nanjing, Peoples R China

2.Guotai Taizhou Ctr Technol Innovat Vet Biol, Taizhou, Peoples R China

3.Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou, Peoples R China

4.Nanjing Agr Univ, Coll Vet Med, Nanjing, Peoples R China

5.Shandong Vocat Anim Sci & Vet Coll, Weifang, Peoples R China

关键词： pseudorabies virus; attenuation; immunogenicity; codon deoptimization; US3-S; UL56

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.2； 五年影响因子：6.2 ）

ISSN：

年卷期： 2023 年 14 卷

页码：

收录情况： SCI

摘要： Since 2011, pseudorabies based on the pseudorabies virus (PRV) variant has emerged as a serious health issue in pig farms in China. The PRV gE/TK or gE/gI/TK deletion strains protect against emerging PRV variants. However, these variants may cause lethal infections in newborn piglets without PRV antibodies. Previous studies have shown that codon deoptimization of a virulence gene causes virus attenuation. Accordingly, we deoptimized US3-S (US3 gene encoding a short isoform that represents approximately 95% of the total US3 transcription) and UL56 genes (first 10 or all codons) of PRV gE/TK deletion strain (PRV Delta TK&gE-AH02) to generate six recombinant PRVs through bacterial artificial chromosome technology. In swine testicular cells, recombinant PRVs with all codon deoptimization of US3-S or UL56 genes were grown to lower titers than the parental virus. Notably, US3-S or UL56 with all codon deoptimization reduced mRNA and protein expressions. Subsequently, the safety and immunogenicity of recombinant PRVs with codon deoptimization of US3-S or UL56 are evaluated as vaccine candidates in mice and piglets. The mice inoculated with recombinant PRVs with codon deoptimization of US3-S or UL56 showed exceptional survival ability without severe clinical signs. All codons deoptimized (US3-S and UL56) significantly decreased virus load and attenuated pathological changes in the brains of the mice. Moreover, the protection efficiency offered by recombinant PRVs with codon deoptimization of US3-S or UL56 showed similar effects to PRV Delta TK&gE-AH02. Remarkably, the 1-day-old PRV antibody-negative piglets inoculated with PRV Delta TK&gE-US3-ST-CD (a recombinant PRV with all codon deoptimization of US3-S) presented no abnormal clinical symptoms, including fever. The piglets inoculated with PRV Delta TK&gE-US3-ST-CD showed a high serum neutralization index against the PRV variant. In conclusion, these results suggest using codon deoptimization to generate innovative live attenuated PRV vaccine candidates.

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