Bai Yu, Chen Weiye, Tong Tiegang, Zhang Weijun, Xu Shulan, Wang Qun, Sun Qingge, Liu Guangliang, Bu Zhigao, Wu Donglai
National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, CAAS, Harbin 150001, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Jul;24(7):1258-62.
Equine interferon-gamma (eIFN-gamma) expressed both in E. coli and baculovirus were evaluated for antiviral activity against recombinant Vesicular Stomatits Virus expressing green fluorescence protein (rVSV-GFP) in EFK-78 cells. The assays were conducted in 96-well plate. Virus infectivity was measured by quantifying GFP-positive cells, instead of quantifying the CPE reduction. Prior to infection of EFK-78 cells with rVSV-GFP, the cells were incubated with eIFN-gamma. 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 titers of antiviral activity were 1 x 10(3) AU/mL and 1 x 10(5) AU/mL of eIFN-gamma expressed from E. coli and baculovirus, respectively. The antiviral activities of the recombinant eIFN-gamma were highly efficient and specific, as it was blocked by mAbs against eIFN-gamma.
对在大肠杆菌和杆状病毒中表达的马γ干扰素（eIFN-γ）针对在EFK-78细胞中表达绿色荧光蛋白的重组水疱性口炎病毒（rVSV-GFP）的抗病毒活性进行了评估。实验在96孔板中进行。通过对GFP阳性细胞进行定量来测量病毒感染性，而不是通过定量细胞病变效应（CPE）的减少。在用rVSV-GFP感染EFK-78细胞之前，将细胞与eIFN-γ一起孵育。与未处理的细胞相比，用eIFN-γ处理的细胞中EFK-78细胞中的GFP表达以剂量依赖性方式显著降低。从大肠杆菌和杆状病毒表达的eIFN-γ的抗病毒活性滴度分别为1×10³ AU/mL和1×10⁵ AU/mL。重组eIFN-γ的抗病毒活性高效且特异，因为它被抗eIFN-γ的单克隆抗体阻断。
