Forloni Gianluigi, Iussich Selina, Awan Tazeen, Colombo Laura, Angeretti Nadia, Girola Laura, Bertani Ilaria, Poli Giorgio, Caramelli Maria, Grazia Bruzzone Maria, Farina Laura, Limido Lucia, Rossi Giacomina, Giaccone Giorgio, Ironside James W, Bugiani Orso, Salmona Mario, Tagliavini Fabrizio
Istituto di Ricerche Farmacologiche Mario Negri, Istituto Nazionale Neurologico Carlo Besta, and Istituto di Microbiologia e Immunologia Veterinaria, Università degli Studi, 20100 Milano, Italy.
Proc Natl Acad Sci U S A. 2002 Aug 6;99(16):10849-54. doi: 10.1073/pnas.162195499. Epub 2002 Jul 29.
Prion diseases are transmissible neurodegenerative disorders of humans and animals for which no effective treatment is available. Conformationally altered, protease-resistant forms of the prion protein (PrP) termed PrP(Sc) are critical for disease transmissibility and pathogenesis, thus representing a primary target for therapeutic strategies. Based on previous findings that tetracyclines revert abnormal physicochemical properties and abolish neurotoxicity of PrP peptides in vitro, we tested the ability of these compounds to interact with PrP(Sc) from patients with the new variant of Creutzfeldt-Jakob disease (vCJD) and cattle with bovine spongiform encephalopathy (BSE). The incubation with tetracycline hydrochloride or doxycycline hyclate at concentrations ranging from 10 microM to 1 mM resulted in a dose-dependent decrease in protease resistance of PrP(Sc). This finding prompted us to investigate whether tetracyclines affect prion infectivity by using an animal model of disease. Syrian hamsters were injected intracerebrally with 263K scrapie-infected brain homogenate that was coincubated with 1 mM tetracycline hydrochloride, 1 mM doxycycline hyclate, or vehicle solution before inoculation. Hamsters injected with tetracycline-treated inoculum showed a significant delay in the onset of clinical signs of disease and prolonged survival time. These effects were paralleled by a delay in the appearance of magnetic-resonance abnormalities in the thalamus, neuropathological changes, and PrP(Sc) accumulation. When tetracycline was preincubated with highly diluted scrapie-infected inoculum, one third of hamsters did not develop disease. Our data suggest that these well characterized antibiotics reduce prion infectivity through a direct interaction with PrP(Sc) and are potentially useful for inactivation of BSE- or vCJD-contaminated products and prevention strategies.
朊病毒疾病是人和动物的可传播性神经退行性疾病,目前尚无有效的治疗方法。构象改变且对蛋白酶具有抗性的朊病毒蛋白(PrP)形式,即PrP(Sc),对于疾病的传播和发病机制至关重要,因此是治疗策略的主要靶点。基于先前的研究结果,即四环素在体外可逆转PrP肽的异常物理化学性质并消除其神经毒性,我们测试了这些化合物与新型克雅氏病(vCJD)患者和牛海绵状脑病(BSE)牛的PrP(Sc)相互作用的能力。用浓度范围为10 microM至1 mM的盐酸四环素或强力霉素进行孵育,导致PrP(Sc)对蛋白酶的抗性呈剂量依赖性降低。这一发现促使我们通过疾病动物模型研究四环素是否影响朊病毒的感染性。在接种前,将叙利亚仓鼠脑内注射263K羊瘙痒病感染的脑匀浆,该匀浆与1 mM盐酸四环素、1 mM强力霉素或溶剂溶液共同孵育。注射了四环素处理接种物的仓鼠在疾病临床症状出现方面有显著延迟,存活时间延长。这些效应与丘脑磁共振异常的出现延迟、神经病理学变化和PrP(Sc)积累平行。当四环素与高度稀释的羊瘙痒病感染接种物预孵育时,三分之一 的仓鼠未发病。我们的数据表明,这些特性明确的抗生素通过与PrP(Sc)直接相互作用降低朊病毒感染性,可能有助于灭活受BSE或vCJD污染的产品及预防策略。