Krasemann S, Groschup M, Hunsmann G, Bodemer W
German Primate Centre, Department of Virology and Immunology, Göttingen, Germany.
J Immunol Methods. 1996 Dec 15;199(2):109-18. doi: 10.1016/s0022-1759(96)00165-2.
Prion diseases are 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). To generate monospecific antisera against human prion proteins we have immunized mice with DNA coding for different human prion proteins. We constructed immunization vectors expressing individual genotypes of either the cellular prion gene (PRNP) or mutant forms under appropriate promoters. This approach avoids the preparation of infectious material for immunization. To circumvent immunological tolerance prion protein-deficient PrP0/0 mice were used for the DNA-mediated immunization. Thereby monospecific sera were raised capable of specifically precipitating in vitro synthesized human prion proteins. With prion protein-specific peptide ELISAs, we found that antibodies are predominantly directed against the octapeptide repeat region and to a lesser extent to regions comprising the signal peptide, the neurotoxic domain or the GPI anchor. In contrast, prion gene-positive (PrP+/+) BALB/c mice immunized under the same experimental conditions as the PrP0/0 mice did not respond with antibody formation against the human prion protein. This is the first report clearly showing that immune competent prion protein-deficient mice react with a vigorous polyclonal immune response after DNA-mediated immunization with human prion gene sequences.
朊病毒疾病是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）。为了产生针对人类朊病毒蛋白的单特异性抗血清，我们用编码不同人类朊病毒蛋白的DNA免疫小鼠。我们构建了在适当启动子下表达细胞朊病毒基因（PRNP）的个体基因型或突变形式的免疫载体。这种方法避免了制备用于免疫的感染性物质。为了规避免疫耐受，使用朊病毒蛋白缺陷的PrP0/0小鼠进行DNA介导的免疫。由此产生了能够特异性沉淀体外合成的人类朊病毒蛋白的单特异性血清。通过朊病毒蛋白特异性肽ELISA，我们发现抗体主要针对八肽重复区域，在较小程度上针对包含信号肽、神经毒性结构域或糖基磷脂酰肌醇锚的区域。相比之下，在与PrP0/0小鼠相同的实验条件下免疫的朊病毒基因阳性（PrP+/+）BALB/c小鼠没有产生针对人类朊病毒蛋白的抗体形成反应。这是第一份清楚表明具有免疫能力的朊病毒蛋白缺陷小鼠在用人类朊病毒基因序列进行DNA介导的免疫后会产生强烈的多克隆免疫反应的报告。
