Bioassay for human prions with the follicular dendritic cells

用滤泡树突状细胞对人朊病毒进行生物测定

• 批准号: 12557059
• 负责人: KITAMOTO Tetsuyuki
• 金额: $ 8.26万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557059/
• 关键词: Creutzfeldt-Jakob disease; prions; knock-in mouse; transgenic mouse; follicular dedritic cells

The purpose of this study is to establish a new bioassay for human prion infection using abnormal prion protein (PrP^<Sc>) accumulations in the follicular dendritic cells (FDC). This year, I analyzed PrP^<Sc> accumulations in FDC both in the transgenic mouse and the knock-in mouse. The both mouse models demonstrated a short incubation periond (〜150 days) by the intracerebral inoculation of human prions from a patient with sporadic Creutzfeldt-Jakob disease. The knock-in mouse had PrP^<Sc> accumulations in the FDC, but the transgenic mouse did not. This phenomenon is due to an attenuated expression of the recombinant PrP in the FDC using our transgenic promoter. With the knock-in mouse, PrP^<Sc> accumulations were detected within 30 days post-inoculation. With dilution series, FDC bioassay system, which takes 75 days, showed enough sensitivity (50% positive in 10-7 diluted human prions) to compare the previous intracerebral inoculation bioassay, which takes 2 years incubation. Therefore, FDC assay provides a rapid and sensitive bioassay for human prion infections. This system is also available to assay human prions of variant CJD (vCJD) in United Kingdom. All the knock-in mice showed the positive FDC stainings with vCJD prions inoculations after 75 post -inoculation.

本研究的目的是建立一种新的利用滤泡树突状细胞（FDC）中异常朊蛋白（PrP^）积累来检测人朊病毒感染的生物学方法<Sc>。今年，我分析了PrP^<Sc>在转基因小鼠和基因敲入小鼠FDC中的积累。通过脑内接种来自散发性Creutzfeldt-Jakob病患者的人朊病毒，两种小鼠模型均显示出较短的潜伏期（约150天）。敲入小鼠在FDC中有PrP^<Sc>积累，但转基因小鼠没有。这种现象是由于使用我们的转基因启动子在FDC中减弱了重组PrP的表达。对于基因敲入小鼠，在<Sc>接种后30天内检测到PrP^积累。使用稀释系列，FDC生物测定系统（需要75天）显示出足够的灵敏度（在10-7稀释的人朊病毒中为50%阳性），可与之前的脑内接种生物测定（需要2年孵育）进行比较。因此，FDC检测为人类朊病毒感染提供了快速和灵敏的生物检测。该系统在英国也可用于检测变异型CJD（vCJD）的人朊病毒。所有基因敲入小鼠在接种vCJD朊病毒后75天均显示阳性FDC染色。

项目成果

Jun Tateishi, Tetsuyuki Kitamoto, Shirou Mohri, Sakae Satoh, Tetsuo Sato, Ailsa Shepherd and Malcolm R: "Macnaughton Scrapie Removal using Planova Virus Removal Filter"Biologicals. 29. 17-25 (2001)

Jun Tateishi、Tetsuyuki Kitamoto、Shirou Mohri、Sakae Satoh、Tetsuo Sato、Ailsa Shepherd 和 Malcolm R：“使用 Planova 病毒去除过滤器去除麦克诺顿瘙痒症”生物制品。

Konaka K, Kaido M, Okuda Y, Aoike F, Abe K, Kitamoto T, Yanagihara T: "Proton magnetic resonance spectroscopy of a patient with Gerstmann-Straussler-Scheinker disease"Neuroradiology. 42(9). 662-665 (2000)

Konaka K、Kaido M、Okuda Y、Aoike F、Abe K、Kitamoto T、Yanagihara T：“Gerstmann-Straussler-Scheinker 病患者的质子磁共振波谱”神经放射学。

Mariko Yamashita. Toru Yamamoto, Kazuto Nishinaka, Fukashi: "Severe brain atrophy in a case of thalamic variant of sporadic CJD with plaque-like PrP deposition"Neuropathology. 21. 138-143 (2001)

山下真理子。

Nakamura Y, Yanagawa H, Kitamoto T, Sato T: "Epidemiologic features of 65 Creutzfeldt-Jakob disease patients with a history of cadaveric dura mater transplantation in Japan"Epidemiol Infect.. 125(1). 201-205 (2000)

Nakamura Y、Yanakawa H、Kitamoto T、Sato T：“日本 65 例有尸体硬脑膜移植史的克雅氏病患者的流行病学特征”流行病感染.. 125(1)。

Jun Tateishi, Tetsuyuki Kitamoto, Shirou Mohri, Sakae Satoh, Tetsuo Sato, Ailsa Shepherd, Malcolm R.Macnaughton: "Scrapie Removal using Planova Virus Removal Filters"Biologicals. 29. 17-25 (2001)

Jun Tateishi、Tetsuyuki Kitamoto、Shirou Mohri、Sakae Satoh、Tetsuo Sato、Ailsa Shepherd、Malcolm R.Macnaughton：“使用 Planova 病毒去除过滤器去除瘙痒病”生物制品。