Keirstead Natalie D, Lee Changhee, Yoo Dongwan, Brooks Andrew S, Hayes M Anthony
Department of Pathobiology, University of Guelph, Guelph, Ontario, Canada.
Antiviral Res. 2008 Jan;77(1):28-38. doi: 10.1016/j.antiviral.2007.08.001. Epub 2007 Aug 27.
Ficolins are collagenous lectins that bind N-acetylated glycans and participate in innate immune responses, including phagocytosis and complement activation. Related collagenous lectins such as mannan binding lectin (MBL) and surfactant proteins A and D possess antiviral activity, but this activity has not been demonstrated for ficolins. In these studies, we used purified porcine plasma ficolin alpha and recombinant ficolin alpha to assess their ability to bind and neutralize porcine reproductive and respiratory virus (PRRSV) in various assays. Recombinant ficolin alpha was designed with a C-terminal 6-histidine tag using a pcDNA3.1 expression vector system in CHO K1 cells. Plasma-purified and recombinant ficolin alpha reduced cytopathic effect of PRRSV-infected Marc-145 cells in neutralization assays and inhibited replication of infectious viral particles in a GlcNAc-dependent manner. In vitro replication determined by plaque assay was inhibited in the presence of plasma-purified ficolin alpha and recombinant ficolin. Immunoreactive plasma ficolin alpha and recombinant ficolin alpha also bound PRRSV-coated wells in a GlcNAc-dependent manner. These studies indicate that porcine ficolin can bind and neutralize a common arterivirus that is a major pathogen of swine.
纤维胶凝蛋白是一种胶原凝集素，可结合N - 乙酰化聚糖并参与先天免疫反应，包括吞噬作用和补体激活。相关的胶原凝集素，如甘露糖结合凝集素（MBL）以及表面活性蛋白A和D具有抗病毒活性，但纤维胶凝蛋白尚未证实有此活性。在这些研究中，我们使用纯化的猪血浆纤维胶凝蛋白α和重组纤维胶凝蛋白α，通过各种试验评估它们结合和中和猪繁殖与呼吸综合征病毒（PRRSV）的能力。使用pcDNA3.1表达载体系统在CHO K1细胞中设计了带有C末端6 - 组氨酸标签的重组纤维胶凝蛋白α。在中和试验中，血浆纯化的和重组的纤维胶凝蛋白α降低了PRRSV感染的Marc - 145细胞的细胞病变效应，并以N - 乙酰葡糖胺依赖的方式抑制了传染性病毒颗粒的复制。在血浆纯化的纤维胶凝蛋白α和重组纤维胶凝蛋白存在的情况下，通过空斑试验测定的体外复制受到抑制。具有免疫反应性的血浆纤维胶凝蛋白α和重组纤维胶凝蛋白α也以N - 乙酰葡糖胺依赖的方式结合PRRSV包被的孔。这些研究表明，猪纤维胶凝蛋白可以结合并中和一种常见的动脉炎病毒，该病毒是猪的主要病原体。
