Neurocenter, Department of Clinical Research and Veterinary Public Health, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
Int J Exp Pathol. 2012 Aug;93(4):259-68. doi: 10.1111/j.1365-2613.2012.00821.x.
Central nervous system (CNS) infections in ruminant livestock, such as listeriosis, are of major concern for veterinary and public health. To date, no host-specific in vitro models for ruminant CNS infections are available. Here, we established and evaluated the suitability of organotypic brain-slices of ruminant origin as in vitro model to study mechanisms of Listeria monocytogenes CNS infection. Ruminants are frequently affected by fatal listeric rhombencephalitis that closely resembles the same condition occurring in humans. Better insight into host-pathogen interactions in ruminants is therefore of interest, not only from a veterinary but also from a public health perspective. Brains were obtained at the slaughterhouse, and hippocampal and cerebellar brain-slices were cultured up to 49 days. Viability as well as the composition of cell populations was assessed weekly. Viable neurons, astrocytes, microglia and oligodendrocytes were observed up to 49 days in vitro. Slice cultures were infected with L. monocytogenes, and infection kinetics were monitored. Infected brain cells were identified by double immunofluorescence, and results were compared to natural cases of listeric rhombencephalitis. Similar to the natural infection, infected brain-slices showed focal replication of L. monocytogenes and bacteria were predominantly observed in microglia, but also in astrocytes, and associated with axons. These results demonstrate that organotypic brain-slice cultures of bovine origin survive for extended periods and can be infected easily with L. monocytogenes. Therefore, they are a suitable model to study aspects of host-pathogen interaction in listeric encephalitis and potentially in other neuroinfectious diseases.
反刍动物中枢神经系统（CNS）感染，如李斯特菌病，对兽医和公共卫生具有重大意义。迄今为止，尚无针对反刍动物 CNS 感染的特定宿主体外模型。在这里，我们建立并评估了源自反刍动物的器官型脑切片作为体外模型研究李斯特菌 CNS 感染机制的适用性。反刍动物经常受到致命李斯特菌脑脊髓炎的影响，这种疾病与人类中发生的相同疾病非常相似。因此，更好地了解反刍动物中的宿主-病原体相互作用不仅从兽医的角度，而且从公共卫生的角度来看都是很有意义的。在屠宰场获得大脑，并培养海马和小脑脑切片，最长可达 49 天。每周评估细胞群体的活力和组成。在体外可观察到存活的神经元、星形胶质细胞、小胶质细胞和少突胶质细胞长达 49 天。用李斯特菌感染切片培养物，并监测感染动力学。通过双重免疫荧光鉴定感染的脑细胞，并将结果与李斯特菌脑脊髓炎的自然病例进行比较。与自然感染相似，感染的脑切片显示李斯特菌的局灶性复制，细菌主要存在于小胶质细胞中，但也存在于星形胶质细胞中，并与轴突相关。这些结果表明，源自牛的器官型脑切片培养物可长期存活，并且容易被李斯特菌感染。因此，它们是研究李斯特菌性脑炎和潜在其他神经感染性疾病中宿主-病原体相互作用方面的合适模型。
