Department of Animal Sciences and Aquatic Ecology, Faculty of Bioscience Engineering, Ghent University, 9000, Ghent, Belgium.
Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent University, 9000, Ghent, Belgium.
Vet Res. 2019 Oct 1;50(1):75. doi: 10.1186/s13567-019-0694-3.
Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a foodborne pathogen which causes illness in humans. Ruminants are the main reservoirs and EHEC predominantly colonizes the epithelium of the recto-anal junction of cattle. Immunosuppression by EHEC promotes re-infection of cattle. However, bovine lactoferrin (bLF) apparently can overrule the immunosuppression by inducing EHEC-specific IgA responses at the mucosal site. The IgA responses are significantly correlated with reduced EHEC shedding and the absence of colonization at the rectal mucosa following re-infection. Therefore, to examine the interaction between bLF and bovine rectal epithelial cells, we first developed a method to establish a primary cell culture of epithelial cells of the rectum of cattle. Furthermore, we used LC-MS/MS to demonstrate the presence of secreted lactoferrin in bovine milk and the absence of a "delta" isoform which is known to translocate to the nucleus of cells. Nevertheless, lactoferrin derived from bovine milk was internalized by rectal epithelial cells and translocated to the nuclei. Moreover, nuclear translocation of bLF was significantly enhanced when the epithelial cells were inoculated with EHEC, as demonstrated by confocal fluorescence microscopy and confirmed by Raman microscopy and 3D imaging.
肠出血性大肠杆菌（EHEC）O157：H7 是一种食源性病原体，可导致人类患病。反刍动物是主要的宿主，EHEC 主要定植在牛的直肠-肛门交界处的上皮细胞。EHEC 的免疫抑制作用促进了牛的再次感染。然而，牛乳铁蛋白（bLF）显然可以通过在黏膜部位诱导 EHEC 特异性 IgA 反应来克服免疫抑制。IgA 反应与减少 EHEC 脱落和直肠黏膜再次感染后无定植显著相关。因此，为了研究 bLF 和牛直肠上皮细胞之间的相互作用，我们首先开发了一种建立牛直肠上皮细胞原代细胞培养的方法。此外，我们使用 LC-MS/MS 证明了牛乳中分泌乳铁蛋白的存在，并且不存在已知易位到细胞核的“δ”同工型。然而，来自牛乳的乳铁蛋白被直肠上皮细胞内化，并易位到细胞核。此外，通过共聚焦荧光显微镜观察到，当上皮细胞接种 EHEC 时，bLF 的核易位显著增强，并通过拉曼显微镜和 3D 成像得到证实。
Zotero 插件
