重组马g-干扰素抗病毒活性测定及单克隆抗体抑制活性鉴定

Antiviral Activity Determination of Recombinant Equine Interferon-gamma and Identification Inhibited Antiviral Activity of Monoclonal Antibodies

摘要:

为了测定大肠杆菌和杆状病毒表达的重组马g-干扰素是否具有抗病毒活性, 利用这两种干扰素处理马胎肾细胞(EFK-78), 然后接种表达绿色荧光蛋白(GFP)的重组水泡性口炎病毒(VSV*GFP), 观察干扰素对病毒表达GFP的抑制, 测出其抗病毒活性单位分别为1×103 AU/mL、1×105 AU/mL。评价了制备的九株抗重组马g-干扰素单克隆抗体是否可抑制重组马g-干扰素抗病毒活性, 证实其中一株可中和重组马g-干扰素的抗病毒活性。结果表明: 杆状病毒表达的马g-干扰素具有较高的抗病毒活性, 其活性可被一株制备的抗重组马g-干扰素单克隆抗体抑制; 首次获得原核表达的具有抗病毒活性的马g-干扰素。

Abstract:

Equine interferon-gamma (eIFN-g) 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-g. The GFP expression in the EFK-78 cells dramatically decreased in the cells treated with eIFN-g in a dose-dependent manner, comparing with the mock-treated cells. The titers of antiviral activity were 1×103AU/mL and 1×105AU/mL of eIFN-g expressed from E. coli and baculovirus, respectively. The antiviral activities of the recombinant eIFN-g were highly efficient and specific, as it was blocked by mAbs against eIFN-g.