Establishment of an immunoscreening system using recombinant VP1 protein for the isolation of a monoclonal antibody that blocks JC virus infection.
Polyomavirus JC (JCV) infection causes the fatal human demyelinating disease, progressive multifocal leukoencephalopathy. Although the initial interaction of JCV with host cells occurs through direct binding of the major viral capsid protein (VP1) with cell-surface molecules possessing sialic acid, these molecules have not yet been identified. In order to isolate monoclonal antibodies which inhibit attachment of JCV, we established an immunoscreening system using virus-like particles consisting of the VP1. Using this system, among monoclonal antibodies against the cell membrane fraction from JCV-permissive human neuroblastoma IMR-32 cells, we isolated a monoclonal antibody designated as 24D2 that specifically inhibited attachment and infection of JCV to IMR-32 cells. The antibody 24D2 recognized a single molecule of around 60 kDa in molecular weight in the IMR-32 membrane fraction. Immunohistochemical staining with 24D2 demonstrated immunoreactivity in the cell membrane of JCV-permissive cell lines and glial cells of the human brain. These results suggested that the molecule recognized by 24D2 plays a role in JCV infection, and that it might participate as a receptor or a co-receptor in JCV attachment and entry into the cells.
Henmi
Chizuka
C
Laboratory of Molecular and Cellular Pathology, Hokkaido University School of Medicine, CREST, JST, Sapporo 060-8638, Japan.
Sawa
Hirofumi
H
Iwata
Hiroshi
H
Orba
Yasuko
Y
Tanaka
Shinya
S
Nagashima
Kazuo
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification* / Q000494:pharmacology; D001921:Brain / Q000276:immunology / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D007577:JC Virus / Q000187:drug effects / Q000502:physiology*; D009457:Neuroglia / Q000276:immunology / Q000378:metabolism; D027601:Polyomavirus Infections / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014412:Tumor Virus Infections / Q000276:immunology / Q000517:prevention & control / Q000821:virology; D014771:Virion / Q000187:drug effects / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D011994:Recombinant Proteins; C062481:VP1 protein, polyomavirus; D016650:Fluorescein-5-isothiocyanate