Transmission Experiment with transgenic model

转基因模型传播实验

• 批准号: 09557056
• 负责人: KITAMOTO Tetsuyuki
• 金额: $ 7.87万
• 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: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557056/
• 关键词: prion disease; prion protein; transgenic mice; gene replacement; transmission experiment; humanized mice

To make a sensitive bioassay system for human prions, we established the transgenic mice with human/mouse chimeric prion protein. We have 3 transgenic lines with codon 129 Met prion protein, and 2 lines with codon 129 Val. Transgenic lines showed various expression level of recombinant prion protein from 0.6 times to 18 times, comparing with the expression of wild-type mouse. The low expression lines (0.6 or 1.2 times) with ablated background showed the shortest incubation period (approximately 150 days) after the intracerebral inoculation of the brain tissues from a patient with sporadic Creutzfeldt-Jakob disease (CJD) (wild type : codon 129 Met/Met, codon 219 Glu/Glu), and the medium expression lines (2 or 4 times) showed the long incubation period. The high expression lines (9 or 18 times) were seen to be inadequate for the transmission experiment because of showing the high expression syndrome and longer incubation periods. Next, we did the transmission experiment form patients with sporadic CJD (codon 129 Val/Met genotype), dura-associated CJD (codon 129 Met/Met genotype). Among these transmission studies, we recognized that a special type of dura-associated CJD showed unsuccessful transmission even after 360 days' incubation. Therefore, not only with the clinicopathological changes, but also with the transmission experiment, we identified a distinct type of dura-associated CJD with many florid plaques as seen in patients with new variant CJD in United Kingdom.

为了建立一个敏感的人朊病毒生物检测系统，我们建立了人/鼠嵌合朊病毒转基因小鼠。我们获得了3个密码子129 Met朊蛋白的转基因株系和2个密码子129瓦尔的转基因株系。转基因小鼠的重组朊蛋白表达量是野生型小鼠的0.6 ~ 18倍。将散发性克雅氏病（CJD）（野生型：密码子129 Met/Met，密码子219 Glu/Glu）患者脑组织接种于脑内，背景消除的低表达株（0.6或1.2倍）显示出最短的潜伏期（约150天），而中表达株（2或4倍）显示出最长的潜伏期。高表达株系（9或18倍）由于表现出高表达综合征和较长的潜伏期，被认为不适合进行传递实验。其次，我们对散发性CJD（密码子129瓦尔/Met基因型）、硬脑膜相关性CJD（密码子129 Met/Met基因型）患者进行了传播实验。在这些传播研究中，我们认识到一种特殊类型的硬脑膜相关的克雅氏病即使在360天的潜伏期后也显示不成功的传播。因此，不仅与临床病理学变化，而且与传播实验，我们确定了一个独特的类型的硬脑膜相关的CJD与许多华丽的斑块中看到的新变异型CJD患者在英国。

项目成果

Yamada H.,Itoh Y.,Inaba A.,Wada Y.,Takashima M.,Satoh S.,Kamata T.,Okeda R.,Kayano T.,Suematsu N.,Kitamoto T.,Otomo E.,Matsushita M.,Mizusawa H.: "An inherited prion disease with PrP P105L mutation : clinicopathological and PrP heterogeneity"Neurology. 53

山田 H.、伊藤 Y.、稻叶 A.、和田 Y.、高岛 M.、佐藤 S.、蒲田 T.、Okeda R.、茅野 T.、末松 N.、北本 T.、大友 E.、松下 M

Shimizu S.,Hoshi K.,Muramoto T.,Homma M.,Ironside JW,Kusuhara S.,Sato T.,Yamamoto T.,Kitamoto T.: "Creutzfeldt-Jakob disease with florid plaques after cadaveric dural grafting"Arch. Neurol.. 56. 357-362 (1999)

Shimizu S.、Hoshi K.、Muramoto T.、Homma M.、Ironside JW、Kusuhara S.、Sato T.、Yamamoto T.、Kitamoto T.：“尸体硬脑膜移植后伴有华丽斑块的克雅氏病”Arch。

Shimizu S,Hoshi K,Muramoto T,Homma M,Ironside JW,Kuzuhara S,Sato T,Yamamoto T,Kitamoto T.: "Creutzfeldt-Jakob disease with florid plaques after cadaveric dural grafting." Auch.Neurol. (in press). (1998)

Shimizu S、Hoshi K、Muramoto T、Homma M、Ironside JW、Kuzuhara S、Sato T、Yamamoto T、Kitamoto T.：“尸体硬脑膜移植后伴有华丽斑块的克雅氏病。”

Matsuda H.,Mitsuda H.,Nakamura N.,Furusawa S.,Mohri S.,Kitamoto T.: "A chicken monoclonal antibody with specificity for the N-terminal of human prion protein"FEMS Immunol. Med. Microbiol. 23. 189-194 (1999)

Matsuda H.、Mitsuda H.、Nakamura N.、Furusawa S.、Mohri S.、Kitamoto T.：“一种对人朊病毒蛋白 N 末端具有特异性的鸡单克隆抗体”FEMS Nutritionol。

**masaki M.,Oyanagi K.,Mori O.,Ohyama M.,Terashi A.,Kitamoto T.,**ayama Y.: "Variant Gerstmann-Straussler syndrome with the P105L prion protein gene mutation : an unusual case with nigral degeneration and widespread neurofibrillary tangles"Acta Neuropatho

**masaki M.、Oyanagi K.、Mori O.、Ohyama M.、Terashi A.、Kitamoto T.、**ayama Y.：“具有 P105L 朊病毒蛋白基因突变的变种 Gerstmann-Straussler 综合征：一个不寻常的病例