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Enhanced productivity and stability of PRV in recombinant ST-Tret1 cells

文献类型： 外文期刊

作者： Xu, Yue 1 ; Bao, Xi 1 ; Chen, Li 1 ; Zhuang, Tenghan 1 ; Xu, Yang 5 ; Feng, Lei 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Vet Immunol & Engn, Natl Res Ctr Engn & Technol Vet Biol, Nanjing, Jiangsu, Peoples R China

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

3.Minist Sci & Technol, Lab Food Qual & Safety, State Key Lab Cultivat Base, Nanjing, Jiangsu, Peoples R China

4.Jiangsu Univ, Sch Pharm, Zhenjiang, Jiangsu, Peoples R China

5.Shanghai Pharmaceut Sch, Shanghai, Peoples R China

关键词： PRV; Intracellular trehalose; Freezing resistance; Freeze -thaw resistance; Cell -substrate

期刊名称：BIOLOGICALS （ 影响因子：1.7； 五年影响因子：1.6 ）

ISSN： 1045-1056

年卷期： 2023 年 83 卷

页码：

收录情况： SCI

摘要： Productivity and stability of Pseudorabies virus (PRV) are critical for the manufacture and storage of live attenuated pseudorabies vaccine. Trehalose is commonly used as a cryoprotectant to stabilize organisms during freezing and lyophilization. Trehalose transporter 1 (Tret1), derived from Polypedilum vanderplanki, can deliver trehalose with a reversible transporting direction. In this study, we demonstrated that productivity and stability of PRV proliferated in recombinant ST cells with stable expression of Tret1 were enhanced. As a result, a five-fold increase of intracellular trehalose amount was observed, and the significant increase of progeny viral titer was achieved in recombinant cells with the addition of 20 mM trehalose. Particularly, after storage for 8 weeks at 20 degrees C, the loss of viral titer was 0.8 and 1.7 lgTCID50/mL lower than the control group with or without the addition of trehalose. Additionally, the freeze-thaw resistance at -20 degrees C and -70 degrees C of PRV was significantly enhanced. Furthermore, according to standard international protocols, a series of tests, including karyotype analysis, tumorigenicity, and the ability of proliferation PRV, were conducted. Our results demonstrated that the recombinant ST cell with Tret1 is a promising cell substrate and has a high potential for producing more stable PRV for the live attenuated vaccine.

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