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Dose effect on detection of PrP^^ in follicular

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Dose effect on detection of PrP^^ in follicular

dendritic cells of knock-in mice for rapid bioassay

Shirou Mohri , Masahiro Asano , Yukiko Ishikawa , Yuichi Matsuura , Yukitoki Fujita^ and Tetsuyuki Kitamoto^

^Division of Laboratory Animal Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582 Japan ^Division of CJD Science and Technology, Tohoku Uni- versity Graduate School of Medicine

<e-mail> shirou@qda.med.kyushu-u.ac.jp

Abstract

We established a rapid bioassay method for transmissibility of human pri- ons to knock-in mouse expressing human/mouse chimeric prion protein (Ki-ChM mouse). In Ki-ChM mice, we detected accumulation of ab- normal isoform (PrP^^) of prion protein in follicular dendritic cells (FDCs) in lymphoid organ within 14 days following intraperitoneal administration of human prions. Intraperitoneal administration of the inoculum provides an advantage over intracerebral injection, considering only a limited vol- ume of 20|il can be inoculated with the intracerebral route. To overcome this limitation and to confirm dose dependent effect on PrP^^ accumulation with intraperitoneal injection, we made an attempt to inoculate brain ho- mogenate of human sporadic CJD (sCJD) with large dose and repetitive injection of a volume up to

-5000|LI1.

Fifty micro liter of 10"^, 10"^ and 10-^ diluted sCJD were inoculated respective groups intraperitoneally.

Five hundred |il of 10"^ and lO""* dilution of sCJD inoculated with one shot were shown as equivalent infectivity to

50|LI1

of 10'^ and 10"^ dilution of them respectively. Five hundred micro liter of lO""^ dilution of sCJD in- oculated with once a day during 10 days were shown as an equivalent in- fectivity to 50|Lil of 10"^ dilution of it. These results indicate that the de- tection of PrP^'^ in FDCs has increased in proportion to the amount of prion inoculated. It is conceivable that the prion of a low density piles up the intake frequency and the pathogenicity rises as well as a high density prion.

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