Recombinant porcine interferon-gamma expressed in CHO cells and its antiviral activity.

Lingyun Wang; Rongzeng Hao; Yang Yang; Yajun LI; Bingzhou LU; Yuhan Mao; Yue Zhang; Zhenli Gong; Yanhong Liu; Meng QI; Yi RU; Haixue Zheng

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Recombinant porcine interferon-gamma expressed in CHO cells and its antiviral activity.

Author: Lingyun WANG ¹ ; Rongzeng HAO ¹ ; Yang YANG ¹ ; Yajun LI ¹ ; Bingzhou LU ¹ ; Yuhan MAO ¹ ; Yue ZHANG ¹ ; Zhenli GONG ¹ ; Yanhong LIU ¹ ; Meng QI ² ; Yi RU ¹ ; Haixue ZHENG ¹
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1. State Key Laboratory for Animal Disease Control and Prevention, National Foot-and-mouth Disease Reference Laboratory, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China.
2. College of Animal Science and Technology, Tarim University, Alra 843300, Xingjiang, China.
Publication Type:Journal Article
Keywords: Chinese hamster ovarian (CHO) cells; Seneca virus A; antiviral activity; expression and purification of protein; porcine interferon γ
MeSH: Swine; Animals; Cricetinae; Interferon-gamma/pharmacology*; Cricetulus; CHO Cells; Sincalide; Recombinant Proteins/pharmacology*; Antiviral Agents/pharmacology*
From: Chinese Journal of Biotechnology 2023;39(12):4784-4795
CountryChina
Language:Chinese
Abstract: The aim of this study was to produce recombinant porcine interferon gamma (rPoIFN-γ) by Chinese hamster ovarian (CHO) cells expression system and to analyze its antiviral activity. Firstly, we constructed the recombinant eukaryotic expression plasmid pcDNA3.1-PoIFN-γ and transfected into suspension cultured CHO cells for secretory expression of rPoIFN-γ. The rPoIFN-γ was purified by affinity chromatography and identified with SDS-PAGE and Western blotting. Subsequently, the cytotoxicity of rPoIFN-γ was analyzed by CCK-8 test, and the antiviral activity of rPoIFN-γ was evaluated using standard procedures in VSV/PK-15 (virus/cell) test system. Finally the anti-Seneca virus A (SVA) of rPoIFN-γ activity and the induction of interferon-stimulated genes (ISGs) and cytokines were also analyzed. The results showed that rPoIFN-γ could successfully expressed in the supernatant of CHO cells. CCK-8 assays indicated that rPoIFN-γ did not show cytotoxicity on IBRS-2 cells. The biological activity of rPoIFN-γ was 5.59×10⁷ U/mg in VSV/PK-15 system. Moreover, rPoIFN-γ could induced the expression of ISGs and cytokines, and significantly inhibited the replication of SVA. In conclusion, the high activity of rPoIFN-γ was successfully prepared by CHO cells expression system, which showed strong antiviral activity on SVA. This study may facilitate the investigation of rPoIFN-γ function and the development of novel genetically engineered antiviral drugs.