Yin Hua, Li Shaowei, Chai Chunxia, Zhang Fan, Ma Yanhua, Wu Youzhi, Fu Cun, Diao Yun, Zhou Yanyan, Zhang Jinlong, Niu Rui, Wang Wei
State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, 010000, China.
State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, 010000, China; Veterinary Research Institute, Inner Mongolia Academy of Agricultural & Animal Husbandry Sciences, Hohhot, China.
Microb Pathog. 2023 Aug;181:106155. doi: 10.1016/j.micpath.2023.106155. Epub 2023 Jun 8.
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.
I型干扰素具有强大的广谱抗病毒能力和免疫调节功能，其受体几乎在所有类型的细胞中都有表达。牛病毒性腹泻病毒（BVDV）是一种在牛群中造成重大经济损失的重要病原体。在本研究中，构建了携带牛α干扰素（BoIFN-α）基因的重组表达质粒，并将其转化到大肠杆菌BL21（DE3）感受态细胞中。SDS-PAGE和Westernblotting分析表明，重组BoIFN-α蛋白（rBoIFN-α）成功表达。其大小约为36KD，以包涵体形式存在。将变性、纯化和复性后的rBoIFN-α蛋白刺激MDBK细胞后，ISG15、OAS1、IFIT1、Mx1和IFITM1等干扰素刺激基因（ISGs）的表达显著上调，并在12小时达到峰值（P<0.001）。分别以感染复数（moi）为0.1和1.0的BVDV感染MDBK细胞。在用rBoIFN-α蛋白预处理和感染后处理后观察病毒增殖情况。结果表明，变性、纯化和复性后的BoIFN-α蛋白具有良好的生物学活性，能够在体外抑制BVDV在MDBK细胞中的复制，这为BoIFN-α作为抗病毒药物、免疫增强剂以及在BVDV临床应用方面提供了依据。
