Zaborowski Adam, Kordek Radzisław, Botts Gerald T, Liberski Paweł P
I Psychiatric Clinic, Lódź.
Pol J Pathol. 2003;54(1):39-47.
Creutzfeldt-Jakob disease (CJD) in a proportion of cases may have nonspecific clinical signs and symptoms and no characteristic neuroimaging and EEG picture. Thus, neuropathological studies are mandatory for a diagnosis. However, spongiform change, neuronal loss and astrocyte proliferation--the hallmarks of prion diseases, may also be absent or variable. In such cases, the diagnosis should be supported by the detection of prion protein (PrP) by Western blotting or immunohistochemistry (ICC). PrP may not be visualised under "regular" conditions, but it is unmasked following pretreatment procedures: incubation in formic acid or guanidine thiocyanate, microwave treatment, and hydrated or hydrolytic autoclaving, and these methods were included in standard diagnostic procedures in several different protocols. The aim of this study was to compare the effectiveness of these pretreatment methods and to introduce an optimal protocol for our laboratory. For this purpose, we used brain sections of 11 cases of CJD, 1 case of Gerstmann-Sträussler-Scheinker syndrome (GSS), 1 case of kuru and 3 control brains. For pretreatment we used the hydrated and hydrolytic autoclaving and incubation with formic acid. Immunostaining was performed with monoclonal 3F4 antibody against PrP. The best results were achieved with hydrolytic autoclaving. By this procedure we were able to detect the "synaptic" type of PrP accumulation in all CJD cases, as well as in GSS and kuru, while with other two methods the signal was weaker or even absent.
克雅氏病（CJD）在部分病例中可能具有非特异性的临床体征和症状，且没有特征性的神经影像学和脑电图表现。因此，神经病理学研究对于诊断至关重要。然而，海绵状改变、神经元丢失和星形胶质细胞增生——朊病毒病的特征，也可能不存在或表现各异。在这种情况下，应通过蛋白质印迹法或免疫组织化学（ICC）检测朊病毒蛋白（PrP）来支持诊断。在“常规”条件下可能无法检测到PrP，但经过预处理程序后它会显现出来：在甲酸或硫氰酸胍中孵育、微波处理以及水合或水解高压灭菌，这些方法已被纳入几种不同方案的标准诊断程序中。本研究的目的是比较这些预处理方法的有效性，并为我们的实验室引入一种最佳方案。为此，我们使用了11例CJD、1例格斯特曼-施特劳斯勒-谢inker综合征（GSS）、1例库鲁病和3例对照脑的脑切片。预处理时我们采用了水合和水解高压灭菌以及甲酸孵育。用抗PrP的单克隆3F4抗体进行免疫染色。水解高压灭菌取得了最佳效果。通过该程序，我们能够在所有CJD病例以及GSS和库鲁病病例中检测到“突触”型PrP积聚，而使用其他两种方法时信号较弱甚至缺失。
