Suzuki Y, Nakao T, Ito T, Watanabe N, Toda Y, Xu G, Suzuki T, Kobayashi T, Kimura Y, Yamada A
Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Science, Japan.
Virology. 1992 Jul;189(1):121-31. doi: 10.1016/0042-6822(92)90687-k.
An improved binding assay for detection of ganglioside receptors for influenza A, B, and C viruses was developed. In this system, the virions bound to gangliosides that were developed on a silica gel thin-layer plate were detected by mouse monoclonal antibody against viral hemagglutinin and peroxidase-conjugated anti-mouse immunoglobin. No hydrolysis of the gangliosides by viral receptor-destroying enzyme was detected in the present condition. The reactivity of the viruses to gangliosides depended on the amount of developed gangliosides (10 pmols-10 nmols), the molecular species of sialic acid, and their sugar sequences. Human influenza A (PR/8/34), B (Lee/40), and C (Ann Arbor/1/50) viruses bound different receptor epitopes of sialo-sugar chains of gangliosides. The A/PR/8 virus bound most effectively to Neu5Ac-containing lacto-series gangliosides carrying type I and type II sugar chains, followed by ganglio-series and hematoside-series gangliosides. The A/PR/8 virus weakly bound to Neu5Ac alpha 2,6lactotetraosylceramide [IV6(Neu5Ac)Lc4Cer] and Neu5Ac alpha 2,6paragloboside [IV6(Neu5Ac)nLc4Cer] carrying Neu5Ac alpha 2,6Gal sequence, although their Neu5Ac alpha 2,3Gal derivatives were the most potent gangliosides tested. B/Lee/40 bound restrictively to IV6(Neu5Ac)Lc4Cer and IV6(Neu5Ac)nLc4Cer, which carry Neu5Ac alpha 2,6Gal sequence, and type I and type II lacto-series sugar chain, respectively. C/Ann Arbor/1/50 reacted only with 9-O-Ac-Neu5Ac-carrying sugar chains in all the gangliosides tested. This method also allowed the microanalysis of receptor gangliosides of unknown samples. ESK cells, sensitive to the influenza A viruses infection, expressed several kinds of receptor active gangliosides, while those from ESK-R cells, resistant to the virus infection, were undetectable.
开发了一种改进的结合试验，用于检测甲型、乙型和丙型流感病毒的神经节苷脂受体。在该系统中，通过抗病毒血凝素的小鼠单克隆抗体和过氧化物酶偶联的抗小鼠免疫球蛋白检测结合在硅胶薄层板上展开的神经节苷脂上的病毒粒子。在当前条件下未检测到病毒受体破坏酶对神经节苷脂的水解作用。病毒对神经节苷脂的反应性取决于展开的神经节苷脂的量（10皮摩尔 - 10纳摩尔）、唾液酸的分子种类及其糖序列。人类甲型流感病毒（PR/8/34）、乙型流感病毒（Lee/40）和丙型流感病毒（Ann Arbor/1/50）结合神经节苷脂唾液酸糖链的不同受体表位。A/PR/8病毒最有效地结合携带I型和II型糖链的含唾液酸神经氨酸（Neu5Ac）的乳糖系列神经节苷脂，其次是神经节系列和血苷系列神经节苷脂。A/PR/8病毒与携带Neu5Acα2,6Gal序列的Neu5Acα2,6乳糖四糖神经酰胺[IV6(Neu5Ac)Lc4Cer]和Neu5Acα2,6副球蛋白[IV6(Neu5Ac)nLc4Cer]弱结合，尽管它们的Neu5Acα2,3Gal衍生物是测试的最有效的神经节苷脂。B/Lee/40分别限制性地结合携带Neu5Acα2,6Gal序列的IV6(Neu5Ac)Lc4Cer和IV6(Neu5Ac)nLc4Cer以及I型和II型乳糖系列糖链。C/Ann Arbor/1/50仅与所有测试神经节苷脂中携带9 - O - 乙酰基 - 唾液酸（9 - O - Ac - Neu5Ac）的糖链发生反应。该方法还允许对未知样品的受体神经节苷脂进行微量分析。对甲型流感病毒感染敏感的ESK细胞表达几种受体活性神经节苷脂，而对病毒感染具有抗性的ESK - R细胞的受体活性神经节苷脂则无法检测到。
