NR AUEC
AU Tayebi,M.; Enever,P.; Sattar,Z.; Collinge,J.; Hawke,S.
TI Disease-associated prion protein elicits immunoglobulin M responses in vivo
QU Molecular Medicine 2004 Jul-Dec; 10(7-12): 104-11
PT journal article
AB Prion diseases such as Creutzfeldt-Jakob disease are believed to result from the misfolding of a widely expressed normal cellular prion protein, PrPc. The resulting disease-associated isoforms, PrPsc, have much higher beta-sheet content, are insoluble in detergents, and acquire relative resistance to proteases. Although known to be highly aggregated and to form amyloid fibrils, the molecular architecture of PrP9Sc) is poorly understood. To date, it has been impossible to elicit antibodies to native PrPsc that are capable of recognizing PrPsc without denaturation, even in Pm-P(o/o) mice that are intolerant of it. Here we demonstrate that antibodies for native PrPc and PrPsc can be produced by immunization of Pm-P(o/o) mice with partially purified PrPc and PrPsc adsorbed to immunomagnetic particles using high-affinity anti-PrP monoclonal antibodies (mAbs). Interestingly, the polyclonal response to PrPsc was predominantly of the immunoglobulin M (IgM) isotype, unlike the immunoglobulin G (IgG) responses elicited by PrPc or by recombinant PrP adsorbed or not to immunomagnetic particles, presumably reflecting the polymeric structure of disease-associated prion protein. Although heat-denatured PrPsc elicited more diverse antibodies with the revelation of C-terminal epitopes, remarkably, these were also predominantly IgM suggesting that the increasing immunogenicity, acquisition of protease sensitivity, and reduction in infectivity induced by heat are not associated with dissociation of the PrP molecules in the diseased-associated protein. Adsorbing native proteins to immunomagnetic particles may have general applicability for raising polyclonal or monoclonal antibodies to any native protein, without attempting laborious purification steps that might affect protein conformation.
MH Adsorption; Amino Acid Sequence; Animals; Antibodies/immunology; Brain/metabolism; Immune Sera; Immunoglobulin Isotypes; Immunoglobulin M/*immunology; Mice; Microspheres; Molecular Sequence Data; PrPsc Proteins/*immunology/*metabolism; Research Support, Non-U.S. Gov't
AD MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London, UK
SP englisch
PO USA