NR ATHY
AU Hosokawa,T.; Tuchiya,K.; Ueda,S.; Tagawa,Y.; Kimura,K.M.; Zanusso,G.; Casalone,C.; Caramelli,M.; Toniolo,A.; Onodera,T.
TI Comparative analysis of abnormal prion protein in bovine spongiform encephalopathy and Creutzfeldt-Jakob disease by novel monoclonal antibodies
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Cell Biology of PrPc and PrPsc CELL-30
PT Konferenz-Poster
AB
This study was initiated to characterize novel monoclonal antibodies (mAbs) that recognize different epitopes of bovine PrP. Their reactivity was compared by immunohistochemistry and Western blot methods using tissue samples from scrapie-infected mice (Sc-mouse), scrapie-infected sheep (Sc-sheep), CJD, and BSE cases. mAbs directed to conformational epitopes reacted strongly with PrPres forms from all the above samples. In addition, interesting differences were found in the reactivity of two mAbs with the type-1 and the type-2 forms of CJD.
By immunizing PrP knockout mice (Prnp-/-) with synthetic polypeptides, a panel of mAbs directed to bovine PrPc has been developed. The ability of these mAbs to detect abnormal prion proteins in BSE and CJD was studied by immunoblotting and immunohistochemistry methods. All mAbs recognized both normal (PrPc) and abnormal (PrPres) isoforms of PrP in bovine, human and mouse brain tissue. Two mAbs recognizing a conformational PrP epitope were reactive with brain tissue from a broad range of mammals, including mice, humans, cows, and sheep. Of particular interest, was the finding that there were two types of Kuru plaques which was distinguished with the staining pattern by the two new mAbs, 6C4 and T2.
Kuru-type spherical plaques present in the cerebellum of a CJD case with codon 129 Met/Val genotype were stained after formic acid pre-treatment using mAbs T2 (1:2,000) and 6C4 (1:5,000). The staining patterns of plaques were divided into peripheral or central by either mAbS. Positive plaques were revealed predominantly in the cerebellar granular layer with occasional plaques in the molecular layer of type-2 CJD. Synaptic type (type-1) PrPcJD was revealed by both mAbs (T2 and 6C4). Diffuse fine deposition around residual neurons in the affected cortex was seen. In BSE-affected brain, diffuse particulate PrP staining of the neurophile was seen with T2 (1:2,500 dilution) and with 6C4 (1:10,000 dilution). No reaction was detected in tissues of uninfected humans or cows. The distinct staining properties of different anti-PrP mAbs suggest that conversion from PrPc to PrPres does involve multiple steps.
AD Tomoko Hosokawa, Kotaro Tuchiya, Susumu Ueda, Nippon Institute for Builogical Sciences, Tokyo, Japan; Yoichi Tagawa, Kumiko M. Kimura, National Institute of Animal Health, Tsukuba, Japan; Takashi Onodera, Department of Molecular Immunology, Sch. Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan; Gianluigi Zanusso, Department of Neurological Sciences, University of Verona, Verona, Italy; Cristina Casalone, Maria Caramelli, Istituto Zooprofilattico Sperimentale, Torino, Italy; Antonio Toniolo, Department of Clinical and Biological Sciences, University of Insubria, Varese, Italy
SP englisch
PO Deutschland