title: A monoclonal antibody (1D12) defines novel distribution patterns of prion protein (PrP) as granules in nucleus.
abstract: A monoclonal antibody (mAb) panel to bovine prion protein (PrP) was studied by immunoblotting and immunohistochemistry for scrapie and bovine spongiform encephalopathy. A mAb panel recognized both normal (PrP(C)) and abnormal (PrP(Sc)) isoforms of PrP in murine, ovine and bovine brain tissues. Interestingly, an anti-bovine PrP mAb, 1D12, prepared by immunizing PrP gene-knockout mice with a synthetic polypeptides corresponding to codons 153-166 of the bovine PrP gene showed novel patterns of reactivity for prion-uninfected neuronal cells. An enzyme-linked immunosorbent assay-mapping of the mAb epitopes resulted in a reaction of monoclonal 1D12 to YEDRY and M corresponding to amino acids 156-160 and 165 of bovine PrP. Several patterns of bovine PrP(C) distribution in PrP-deficient neuronal cells (HpL3-4) transfected with bovine PrP were observed after different fixation methods. Stained cell surface was observed after formalin fixation by immunofluorescent assay of 1D12 with confocal microscopy, whereas granules in nucleus were stained after acetone fixation. No reactivity in the nucleus was observed to HpL3-4, or HpL3-4mPrP cells expressing mouse PrP. This is the first paper that has reported the detection of the PrP(C) at both cell surface and nuclei of prion-uninfected cell line.
authors: Nippon Institute for Biological Science, Ome 198-0024, Tokyo, Japan.
authors: Tomoko
authors: T
authors: Hosokawa
authors: Kotaro
authors: K
authors: Tsuchiya
authors: Ichiro
authors: I
authors: Sato
authors: Natsumi
authors: N
authors: Takeyama
authors: Susumu
authors: S
authors: Ueda
authors: Yuichi
authors: Y
authors: Tagawa
authors: Kumiko M
authors: KM
authors: Kimura
authors: Izuru
authors: I
authors: Nakamura
authors: Guoying
authors: G
authors: Wu
authors: Akikazu
authors: A
authors: Sakudo
authors: Cristina
authors: C
authors: Casalone
authors: Maria
authors: M
authors: Mazza
authors: Maria
authors: M
authors: Caramelli
authors: Hidehiro
authors: H
authors: Takahashi
authors: Tetsutaro
authors: T
authors: Sata
authors: Katsuaki
authors: K
authors: Sugiura
authors: Andreina
authors: A
authors: Baj
authors: Antonio
authors: A
authors: Toniolo
authors: Takashi
authors: T
authors: Onodera
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002467:Cell Nucleus / Q000378:metabolism*; D007150:Immunohistochemistry; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D011328:Prions / Q000378:metabolism*; D022142:Secretory Vesicles / Q000378:metabolism*; D012756:Sheep; D014018:Tissue Distribution
chemical_list: D000911:Antibodies, Monoclonal; D011328:Prions