Abstract:
Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals that has a significant socio-economic impact. One concern associated with this disease is the ability of its etiological agent, the FMD virus (FMDV), to persist in its hosts through underlying mechanisms that remain to be elucidated. While persistence has been described in cattle and small ruminants, it is unlikely to occur in pigs. One of the factors limiting the progress in understanding FMDV persistence and, in particular, differential persistence is the lack of suitable in vitro models. A primary bovine cell model derived from the dorsal soft palate, which is the primary site of replication and persistence of FMDV in cattle, has been developed, and it seemed relevant to develop a similar porcine model. Cells from two sites of FMDV replication in pigs, namely, the dorsal soft palate and the oropharyngeal tonsils, were isolated and cultured. The epithelial character of the cells from the dorsal soft palate was then assessed by immunofluorescence. The FMDV-sensitivity of these cells was assessed after monolayer infection with FMDV O/FRA/1/2001 Clone 2.2. These cells were also grown in multilayers at the air-liquid interface to mimic a stratified epithelium susceptible to FMDV infection. Consistent with what has been shown in vivo in pigs, our study showed no evidence of persistence of FMDV in either the monolayer or multilayer model, with no infectious virus detected 28 days after infection. The development of such a model opens up new possibilities for the study and diagnosis of FMDV in porcine cells.
摘要:
口蹄疫(FMD)是偶蹄动物的高度传染性病毒性疾病,具有重大的社会经济影响。与这种疾病相关的一个问题是其病因的能力,口蹄疫病毒(FMDV),通过仍有待阐明的潜在机制在其宿主中持续存在。虽然在牛和小反刍动物中已经描述了持久性,它不太可能发生在猪身上。限制FMDV持久性理解进展的因素之一,特别是,差异持久性是缺乏合适的体外模型。源自背侧软腭的原始牛细胞模型,这是FMDV在牛中复制和持续的主要位点,已经开发出来了,这似乎与建立类似的猪模型有关。来自猪FMDV复制的两个位点的细胞,即,背侧软腭和口咽扁桃体,被分离和培养。然后通过免疫荧光评估来自背侧软腭的细胞的上皮特性。用FMDVO/FRA/1/2001克隆2.2单层感染后评估这些细胞的FMDV敏感性。这些细胞也在气-液界面的多层中生长,以模拟对FMDV感染敏感的复层上皮。与在猪体内显示的情况一致,我们的研究表明,没有证据表明FMDV在单层或多层模型中持续存在,感染后28天未检测到感染性病毒。这种模型的发展为猪细胞中FMDV的研究和诊断开辟了新的可能性。