Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China.
Comp Immunol Microbiol Infect Dis. 2010 Jul;33(4):333-42. doi: 10.1016/j.cimid.2008.12.004. Epub 2009 Mar 10.
Interferon gamma (IFN-gamma) is a pleiotropic cytokine that is recognized as an important modulator of the immune response. To date, there is no report that prokaryocyte-derived recombinant equine IFN-gamma has antiviral activity. In this report, the gene coding equine IFN-gamma (EIFN-gamma) mature protein was cloned into pET-28a (+) and the recombinant EIFN-gamma was expressed in Escherichia coli (E. coli). The antiviral activity of expressed recombinant EIFN-gamma was evaluated by using a recombinant Vesicular Stomatitis Virus expressing green fluorescence protein (rVSV-GFP) system in the equine fetal kidney-78 cell line (EFK-78). The GFP expression in the EFK-78 cells dramatically decreased in the cells treated with EIFN-gamma in a dose-dependent manner, comparing with the mock-treated cells. The titer of antiviral activity was 1 x 10(3)AU/ml. These results demonstrated that the EIFN-gamma expressed in this study had good biological activity. Pure forms and sufficient quantities of biologically active IFN-gamma could facilitate the study of its activities in modulating immune responses both in vivo and in vitro.
干扰素 γ（IFN-γ）是一种多功能细胞因子，被认为是免疫反应的重要调节剂。迄今为止，尚无报道表明原核生物来源的重组马干扰素 γ（EIFN-γ）具有抗病毒活性。在本报告中，将编码马 IFN-γ（EIFN-γ）成熟蛋白的基因克隆到 pET-28a（+）中，并在大肠杆菌（E. coli）中表达重组 EIFN-γ。通过在马胎儿肾细胞系（EFK-78）中使用表达绿色荧光蛋白（rVSV-GFP）的重组水疱性口炎病毒（rVSV-GFP）系统评估表达的重组 EIFN-γ的抗病毒活性。与mock 处理的细胞相比，用 EIFN-γ处理的细胞中 GFP 的表达呈剂量依赖性显著降低。抗病毒活性的效价为 1×103 AU/ml。这些结果表明，本研究中表达的 EIFN-γ具有良好的生物学活性。纯形式和足够数量的生物活性 IFN-γ可促进其在体内和体外调节免疫反应的活性研究。
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