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

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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