An establishment of cell line with high sensitivity to rotavirus by transfection of receptor gene and the application for diagnosis in rotavirus infection

为了建立对轮状病毒高敏感性的细胞系，我们测定了禽轮状病毒所有基因组片段的完整序列，并在MA-104细胞上鉴定了禽轮状病毒的受体。此外，我们还试图制备针对轮状病毒受体的单克隆抗体。结果表明：(1)本实验室分离的禽轮状病毒P0-13株11个基因组片段的全核苷酸序列可引出轮状病毒与细胞受体的结合蛋白。然后，克隆编码轮状病毒若干蛋白的基因在猿猴COS7细胞中高水平表达，并检测了所表达蛋白的受体结合功能。, 1995年。病毒学,1995年。Vet.Microbiol。, 1996年。Arch.Virol。, 1996年。病毒基因，1997。(2)通过几种酶处理MA-104细胞单分子，以及轮状病毒与神经节苷类的结合活性，我们认为唾液酸可以作为病毒结合和进入宿主细胞的表面分子。(3)结果表明，鸡蛋清中的卵泡蛋白和牛奶中的粘蛋白能与禽轮状病毒结合，抑制病毒对MA-104细胞的感染。虽然我们制备了许多针对这些粘蛋白的单克隆抗体，但到目前为止，所有的抗体都不能抑制轮状病毒对MA-104细胞的感染。未来，我们将尝试绘制轮状病毒VP8的受体结合区，并合成针对该氨基酸区对应肽的单克隆抗体

In this study, to establish cell lines with high sensitivity to rotavirus, we determined complete sequence of all genomic segments of avian rotavirus, and characterization of a receptor for avian rotavirus on MA-104 cells. Moreover, we attempted to produce monoclonal antibodies directed to receptor for rotavirus. The ressults run as follows :(1) The complete nucleotide sequence of 11 genomic segments of avian rotavirus, P0-13 strain, which was isolated from a pigeon in our laboratory, was determined to elicit binding protein of rotavirus to cellular receptor. Then, the cloning genes encoding several proteins of rotavirus were expressed at a high level in simian COS7 cells and the expressed proteins were examined in the function of receptor binding.(Arch.Virol., 1995. Virology, 1995. Vet.Microbiol., 1996. Arch.Virol., 1996. Virus Genes, 1997. Virus Res., 1997)(2) By the treatment of MA-104 cell monolayrs with several enzymes, and by binding activity of rotaviruses to gangliosides, we suggested that sialic acid served as the surface molecule used by the virus for binding and entry into host cells.(Arch.Virol., in preparation)(3) It was indicated that ovomucin in hen egg white and mucin in cow milk bind to avian rotavirus, and these mucins inhibit infection of the virus to MA-104 cells. Although we produced many of monoclonal antibodies to these mucins, all of the resulting antibodies failed to inhibit infection of rotavirus to MA-104 cells so far.In the future, we will make an attempt to map receptor binding region in VP8 of rotavirus and to produce monoclonal antibodies against the synthetic peptides corresponding to the amino acid region