Abbreviated incubation times for human prions in mice expressing a chimeric mouse-human prion protein transgene
Abbreviated incubation times for human prions in mice expressing a chimeric mouse-human prion protein transgene
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DOI：
10.1073/pnas.2627989100
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发表时间：
2003-04-15
影响因子：
11.1
通讯作者：
Prusiner, SB
中科院分区：
文献类型：
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作者：
Korth, C;Kaneko, K;Prusiner, SB
表达嵌合小鼠-人朊病毒蛋白（PrP）（命名为MHu 2 M）的转基因（Tg）小鼠品系对来自散发性克雅氏病（sCJD）患者的朊病毒易感。为了将孵育时间减少到少于200天，我们构建了转基因，其中MHu 2 M中的9个人类残基中的一个或多个被改变为小鼠。在165和167位具有鼠残基的构建体在Tg（MHu 2 M，M165 V，E167 Q）小鼠中表达，并导致来自sCJD患者的朊病毒的孵育时间缩短至约110天。在96位具有鼠残基的构建体导致sCJD朊病毒的孵育时间延长至280天以上。当鼠残基96、165和167表达时，sCJD朊病毒的缩短孵育时间被废除。变异型克雅氏病朊病毒在Tg（MHu 2 M）小鼠第一代的潜伏期延长至300 - 700天，在Tg（MHu 2 M，M165 V，E167 Q）小鼠的潜伏期约为350天。在Tg（MHu 2 M）小鼠中变异型CJD朊病毒的第二代和第三代，根据孵育时间和蛋白酶抗性、去糖基化PrPSc片段的大小检测到多株朊病毒。我们发现了一个以前未描述的嵌合转基因，缩短了sCJD朊病毒的孵育时间，这将有助于朊病毒物种屏障和人类朊病毒多样性的研究。
Transgenic (Tg) mouse lines that express chimeric mouse-human prion protein (PrP), designated MHu2M, are susceptible to prions from patients with sporadic Creutzfeldt-Jakob disease (sCJD). With the aim of decreasing the incubation time to fewer than 200 days, we constructed transgenes in which one or more of the nine human residues in MHu2M were changed to mouse. The construct with murine residues at positions 165 and 167 was expressed in Tg(MHu2M,M165V,E167Q) mice and resulted in shortening the incubation time to approximate to110 days for prions from sCJD patients. The construct with a murine residue at position 96 resulted in lengthening the incubation time to more than 280 days for sCJD prions. When murine residues 96, 165, and 167 were expressed, the abbreviated incubation times for sCJD prions were abolished. Variant CJD prions showed prolonged incubation times between 300 and 700 days in Tg(MHu2M) mice on first passage and incubation times of approximate to350 days in Tg(MHu2M,M165V,E167Q) mice. On second and third passages of variant CJD prions in Tg(MHu2M) mice, multiple strains of prions were detected based on incubation times and the sizes of the protease-resistant, deglycosylated PrPSc fragments. Our discovery of a previously undescribed chimeric transgene with abbreviated incubation times for sCJD prions should facilitate studies on the prion species barrier and human prion diversity.