Staphylococcus aureus and Escherichia coli cause deviating expression profiles of cytokines and lactoferrin messenger ribonucleic acid in mammary epithelial cells
Staphylococcus aureus and Escherichia coli cause deviating expression profiles of cytokines and lactoferrin messenger ribonucleic acid in mammary epithelial cells
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DOI：
10.3168/jds.2007-0752
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发表时间：
2008-06-01
影响因子：
3.5
通讯作者：
Wellnitz, O.
中科院分区：
文献类型：
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作者：
Griesbeck-Zilch, B.;Meyer, H. H. D.;Wellnitz, O.
病原体入侵乳腺会引起一个复杂的信号网络，激活早期的免疫防御，并导致炎症症状的结果。为了检验乳腺上皮细胞在这些调节和相互作用中的重要性，导致病原体相关的乳房炎过程，我们用实时定量聚合酶链式反应分析了天然免疫系统关键分子的mRNA表达谱。将28头初乳荷斯坦奶牛在哺乳42天分离出乳腺上皮细胞，在标准化条件下分别培养1、6、24 h，分别用革兰氏阳性菌(金黄色葡萄球菌)和革兰氏阴性菌(大肠杆菌)处理1、6、24 h。这两种病原体都增加了与病原体识别有关的蛋白质的mRNA表达模式，如Toll样受体和核因子-kappa B，而革兰氏阴性菌起到了更强的刺激作用。此外，促炎症细胞因子肿瘤坏死因子α、IL-1β、IL-6和趋化因子如IL-8和RANTES(激活后调节，正常T细胞表达和分泌)的表达谱可以证实这一点。值得注意的是，在处理1h后，这些细胞因子和趋化因子在低水平的mRNA表达，在金黄色葡萄球菌中的表达显著高于在受大肠杆菌影响的细胞中的表达。乳铁蛋白的表达模式与病原菌的刺激方式不同(即在1h时，大肠杆菌诱导了较高的mRNA表达，而在金黄色葡萄球菌刺激后24h达到最高水平)。补体因子3是唯一对两种微生物反应相同的测量因子。我们的数据强调了乳腺上皮细胞在乳房免疫防御中的作用，并证实了它们在病原体相关的不同病程乳房炎中的作用。
Pathogens invading the mammary gland cause a complex signaling network that activates the early immune defense and leads to an outcome of inflammation symptoms. To examine the importance of mammary epithelial cells in these regulations and interactions resulting in a pathogen-related course of mastitis, we characterized the mRNA expression profile of key molecules of the innate immune system by quantitative real-time PCR. Mammary gland epithelial cells isolated on d 42 of lactation from 28 first-lactation Holstein dairy cows were cultured separately under standardized conditions and treated for 1, 6, and 24 h with heat-inactivated gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria. Both pathogens increased mRNA expression patterns of proteins involved in pathogen recognition such as Toll-like receptors and nuclear factor-kappa B, whereas gram-negatives acted as a stronger stimulus. Furthermore, this could be confirmed by the expression profile of the proinflammatory cytokines tumor necrosis factor alpha, IL-1 beta, IL-6, and chemokines such as IL-8 and RANTES (regulated upon activation, normal T-cell expressed and secreted). Remarkably, at a low level of mRNA expression after 1 h of treatment these cytokines and chemokines were expressed at a significantly higher level in Staphyloccocus aureus than in Escherichia coli affected cells. Lactoferrin showed a deviating expression pattern to pathogen stimulation (i.e., at the 1-h measuring point Escherichia coli induced a higher mRNA expression, whereas the highest level was reached after 24 h of stimulation with Staphylococcus aureus). Complement factor 3 was the only measured factor that responded equally to both microorganisms. Our data emphasize the role of mammary epithelial cells in the immune defense of the udder and confirm their contribution to pathogen-related different courses of mastitis.