Krasemann S, Groschup M H, Harmeyer S, Hunsmann G, Bodemer W
Department of Virology and Immunology, German Primate Center (DPZ), Goettingen, Germany.
Mol Med. 1996 Nov;2(6):725-34.
Prion diseases belong to a group of neurodegenerative disorders affecting humans and animals. The human diseases include kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome (GSS), and fatal familial insomnia (FFI). The pathogenic mechanisms of the prion diseases are not yet understood. Monoclonal antibodies provide valuable tools in the diagnosis, as well as in the basic research, of several diseases; however, monospecific antisera or monoclonal antibodies (mAbs) against human prion proteins were, until now, not available.
We have developed an immunization protocol based on nucleic acid injection into nontolerant PrP0/0 mice. DNA or RNA coding for different human prion proteins including the mutated sequences associated with CJD, GSS, and FFI were injected into muscle tissue. Mice were primarily inoculated with DNA plasmids encoding the prion protein (PRNP) gene and boosted either with DNA, RNA, or recombinant Semliki Forest Virus particles expressing PRNP. Hybridomas were then prepared.
Different mAbs against human prion proteins were obtained, and their binding behavior was analyzed by peptide enzyme-linked immunosorbent assay, Western blot, immunofluorescence, and immunoprecipitation. Their cross-reactivity with prion protein from other species was also determined. Our mAbs are directed against four different linear epitopes and may also recognize discontinuous regions of the native prion protein.
These antibodies should allow us to address questions concerning the nature of the prion protein as well as the initiation and progression of prion diseases. Moreover, these mAbs can now be used for the diagnosis of prion diseases of humans and animals.
朊病毒疾病属于一组影响人类和动物的神经退行性疾病。人类疾病包括库鲁病、克雅氏病（CJD）、格斯特曼-施特劳斯勒-谢inker综合征（GSS）和致死性家族性失眠症（FFI）。朊病毒疾病的致病机制尚未明确。单克隆抗体为多种疾病的诊断以及基础研究提供了有价值的工具；然而，迄今为止，针对人类朊病毒蛋白的单特异性抗血清或单克隆抗体（mAb）尚未获得。
我们基于将核酸注射到无耐受性的PrP0/0小鼠体内制定了一种免疫方案。将编码不同人类朊病毒蛋白（包括与CJD、GSS和FFI相关的突变序列）的DNA或RNA注射到肌肉组织中。小鼠首先接种编码朊病毒蛋白（PRNP）基因的DNA质粒，然后用DNA、RNA或表达PRNP的重组塞姆利基森林病毒颗粒进行加强免疫。随后制备杂交瘤。
获得了针对人类朊病毒蛋白的不同mAb，并通过肽酶联免疫吸附测定、蛋白质印迹、免疫荧光和免疫沉淀分析了它们的结合行为。还确定了它们与其他物种朊病毒蛋白的交叉反应性。我们的mAb针对四个不同的线性表位，也可能识别天然朊病毒蛋白的不连续区域。
这些抗体应能使我们解决有关朊病毒蛋白的性质以及朊病毒疾病的起始和进展的问题。此外，这些mAb现在可用于诊断人类和动物的朊病毒疾病。
