Sato Yuichiro, Hirayama Makoto, Morimoto Kinjiro, Hori Kanji
Faculty of Pharmacy, Yasuda Women's University, Hiroshima, Japan.
Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Japan.
Methods Mol Biol. 2020;2132:683-693. doi: 10.1007/978-1-0716-0430-4_59.
High mannose (HM)-binding Oscillatoria agardhii agglutinin homologue (OAAH) lectin family is an important class of anti-viral proteins. The OAAH family lectins show potent anti-influenza virus activity with EC of nanomolar levels by binding to HM glycans of the envelope glycoprotein hemagglutinin (HA), thereby inhibiting the viral entry into host cells. No broadly effective neutralizing vaccines for influenza virus are available due to the frequent antigenic drift caused by rapid mutations. Alternatives for vaccines need to be developed to prepare for a possible risk of future emergence of a highly virulent virus. Possible use of antiviral lectins is a simple and useful strategy to prevent viral infection by interfering with the interaction between viral HA and the host sialic acid-containing receptor. High-density glycans of surface HA are primary targets for the lectins to inhibit viral entry. In general, the anti-influenza virus potency of lectins is evaluated by a series of inhibitory assays for infection, such as neutral red dye uptake assay to determine the extent of viral cytopathic effect, and immunofluorescence microscopy to detect the expression of viral proteins in infected cells. Direct interaction between lectins and HA could be evaluated by enzyme-linked immunosorbent assay or surface plasmon resonance analysis.
高甘露糖(HM)结合类颤藻凝集素同源物(OAAH)凝集素家族是一类重要的抗病毒蛋白。OAAH家族凝集素通过与包膜糖蛋白血凝素(HA)的HM聚糖结合,表现出强大的抗流感病毒活性,其半数有效浓度(EC)处于纳摩尔水平,从而抑制病毒进入宿主细胞。由于快速突变导致频繁的抗原漂移,目前尚无针对流感病毒的广泛有效的中和疫苗。需要开发疫苗的替代物,以应对未来可能出现高毒力病毒的风险。使用抗病毒凝集素是一种简单且有用的策略,可通过干扰病毒HA与宿主含唾液酸受体之间的相互作用来预防病毒感染。表面HA的高密度聚糖是凝集素抑制病毒进入的主要靶点。一般来说,凝集素的抗流感病毒效力通过一系列感染抑制试验来评估,如用于确定病毒细胞病变效应程度的中性红染料摄取试验,以及用于检测感染细胞中病毒蛋白表达的免疫荧光显微镜检查。凝集素与HA之间的直接相互作用可通过酶联免疫吸附测定或表面等离子体共振分析来评估。