Distinct immunohistochemical localization in Kuru plaques using novel anti-prion protein antibodies.
By immunizing Prnp-knockout mice with synthetic polypeptides, a panel of mAbs directed to bovine PrP(C) was obtained. The mAb panel was characterized by the ELISA method, where synthetic polypeptides were used for epitope mapping. Different reactivity patterns were identified. The ability of these mAbs to detect abnormal PrP(Sc) in CJD cases was studied by immunohistochemistry. All mAbs were tested for PrP(Sc) in murine, bovine, monkey and human brain tissues. Three mAbs recognized the fragmented PrP epitope in our ELISA. Antibody 1D12 was strongly reactive to ovine and squirrel monkey tissues infected with a scrapie agent, although non-reactive to scrapie-infected mouse tissues. Antibody 2D8 was clearly reactive to type-2 but not type-1 CJD human tissues. Of particular interest was the reactivity of mAb 6C4 with the inner structure of Kuru plaques (peripheral pattern) in a type-2 CJD case and mAb T2, 1D12, 2B11, 2D8, 4B5 and 6G3-2 with the central area (central pattern). The fact that different anti-PrP mAbs possess distinct staining properties suggests that the PrP(c) to PrP(Sc) conversion might involve a multiple-step process.
Hosokawa
Tomoko
T
Nippon Institute for Biological Science, University of Tokyo, Tokyo, Japan.
Ono
Fumiko
F
Tsuchiya
Kotaro
K
Sato
Ichiro
I
Takeyama
Natsumi
N
Ueda
Susumu
S
Zanusso
Gianluigi
G
Takahashi
Hidehiro
H
Sata
Tetsutaro
T
Sakudo
Akikazu
A
Suguira
Katsuaki
K
Baj
Andreina
A
Toniolo
Antonio
A
Yoshikawa
Yasuhiro
Y
Onodera
Takashi
T
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D001921:Brain / Q000473:pathology; D002417:Cattle; D018604:Epitope Mapping; D006801:Humans; D007729:Kuru / Q000473:pathology*; D051379:Mice; D018345:Mice, Knockout; D011328:Prions / Q000032:analysis* / Q000276:immunology; D012453:Saimiri
D000906:Antibodies; D000911:Antibodies, Monoclonal; D011328:Prions