title: Ultrasensitive detection of PrP(Sc) in the cerebrospinal fluid and blood of macaques infected with bovine spongiform encephalopathy prion.
abstract: Prion diseases are characterized by the prominent accumulation of the misfolded form of a normal cellular protein (PrP(Sc)) in the central nervous system. The pathological features and biochemical properties of PrP(Sc) in macaque monkeys infected with the bovine spongiform encephalopathy (BSE) prion have been found to be similar to those of human subjects with variant Creutzfeldt-Jakob disease (vCJD). Non-human primate models are thus ideally suited for performing valid diagnostic tests and determining the efficacy of potential therapeutic agents. In the current study, we developed a highly efficient method for in vitro amplification of cynomolgus macaque BSE PrP(Sc). This method involves amplifying PrP(Sc) by protein misfolding cyclic amplification (PMCA) using mouse brain homogenate as a PrP(C) substrate in the presence of sulfated dextran compounds. This method is capable of amplifying very small amounts of PrP(Sc) contained in the cerebrospinal fluid (CSF) and white blood cells (WBCs), as well as in the peripheral tissues of macaques that have been intracerebrally inoculated with the BSE prion. After clinical signs of the disease appeared in three macaques, we detected PrP(Sc) in the CSF by serial PMCA, and the CSF levels of PrP(Sc) tended to increase with disease progression. In addition, PrP(Sc) was detectable in WBCs at the clinical phases of the disease in two of the three macaques. Thus, our highly sensitive, novel method may be useful for furthering the understanding of the tissue distribution of PrP(Sc) in non-human primate models of CJD.
authors.affiliation: Influenza and Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Murayama
authors.affiliation: Influenza and Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Masujin
authors.affiliation: Influenza and Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
authors.forename: Morikazu
authors.initials: M
authors.lastname: Imamura
authors.affiliation: Chiba Institute of Science Faculty of Risk and Crisis Management, Choshi, Chiba, Japan.
authors.forename: Fumiko
authors.initials: F
authors.lastname: Ono
authors.affiliation: Tsukuba Primate Research Center, National Institute of Biomedical Innovation, Tsukuba, Ibaraki, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shibata
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Tobiume
authors.affiliation: Influenza and Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Yamamura
authors.affiliation: Influenza and Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Shimozaki
authors.affiliation: Tsukuba Primate Research Center, National Institute of Biomedical Innovation, Tsukuba, Ibaraki, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Terao
authors.affiliation: Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Yamakawa
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
mesh_terms: D000818:Animals; D002417:Cattle; D007562:Creutzfeldt-Jakob Syndrome / Q000097:blood / Q000134:cerebrospinal fluid; D004195:Disease Models, Animal; D016643:Encephalopathy, Bovine Spongiform / Q000097:blood* / Q000134:cerebrospinal fluid*; D006801:Humans; D008252:Macaca fascicularis / Q000097:blood* / Q000134:cerebrospinal fluid*; D008297:Male; D051379:Mice; D018620:PrPSc Proteins / Q000097:blood* / Q000134:cerebrospinal fluid*; D014018:Tissue Distribution
chemical_list: D018620:PrPSc Proteins