Biological activity of recombinant bovine IFN-α and inhibitory effect on BVDV in vitro.
Biological activity of recombinant bovine IFN-α and inhibitory effect on BVDV in vitro.
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DOI：
10.2139/ssrn.4352878
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发表时间：
2023-06
影响因子：
3.8
通讯作者：
Hua Yin;Shaowei Li;Chunxia Chai;Fan Zhang;Yanhua Ma;Youzhi Wu;Cun Fu;Yun Diao;Yanyan Zhou;Jinlong Zhang;Niu Rui;Wen Wang
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文献类型：
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作者：
Hua Yin;Shaowei Li;Chunxia Chai;Fan Zhang;Yanhua Ma;Youzhi Wu;Cun Fu;Yun Diao;Yanyan Zhou;Jinlong Zhang;Niu Rui;Wen Wang
I型干扰素具有很强的广谱抗病毒能力和免疫调节功能，其受体在几乎所有类型的细胞中都有表达。牛病毒性腹泻病毒（BVDV）是一种重要的病原体，给牛的生产造成了巨大的经济损失。本研究构建了牛α-干扰素（BoIFN-α）基因的重组表达质粒，并转化E. coliBL 21（DE 3）感受态细胞。SDS-PAGE和Westernblotting分析表明，重组BoIFN-α蛋白（rBoIFN-α）获得了表达。其分子量约为36 KD，以包涵体形式存在。经变性、纯化和复性的rBoIFN-α蛋白刺激MDBK细胞后，ISG 15、OAS 1、IFIT 1、Mx 1和IFITM 1等干扰素刺激基因的表达均显著上调，并在12 h达到高峰（P< 0.001）。MDBK细胞分别以0.1和1.0的moi感染BVDV。经rBoIFN-α蛋白预处理及感染后处理，观察病毒增殖情况。结果表明，经变性、纯化和复性的BoIFN-α蛋白具有良好的生物学活性，在体外能抑制BVDV在MDBK细胞中的复制，为BoIFN-α作为BVDV的抗病毒药物、免疫增强剂和临床应用提供了依据。
Type I interferon has great broad-spectrum antiviral ability and immunomodulatory function, and its receptors are expressed in almost all types of cells. Bovine viral diarrhea virus (BVDV) is an important pathogen causing significant economic losses in cattle. In this study, a recombinant expression plasmid carrying bovine interferon-α(BoIFN-α)gene was constructed and transformed into E. coli BL21 (DE3) competent cells. SDS-PAGE and Westernblotting analysis showed that the recombinant BoIFN-α protein (rBoIFN-α) was successfully expressed. It is about 36KD and exists in the form of inclusion body. When denatured, purified and renatured rBoIFN-α protein stimulated MDBK cells, the expression of interferon stimulating genes (ISGs) such as ISG15, OAS1, IFIT1, Mx1 and IFITM1 were significantly up-regulated, and reached the peak at 12 h (P< 0.001). MDBK cells were infected with BVDV with moi of 0.1 and 1.0, respectively. The virus proliferation was observed after pretreatment with rBoIFN-α protein and post-infection treatment. The results showed that the denatured, purified and renatured BoIFN-α protein had good biological activity and could inhibit the replication of BVDV in MDBK cells in vitro, which provided a basis for BoIFN-α as an antiviral drug, immune enhancer and clinical application of BVDV.