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金黄色葡萄球菌对培养的牛乳腺上皮细胞的黏附作用。

Adherence of Staphylococcus aureus to cultured bovine mammary epithelial cells.

作者信息

Cifrian E, Guidry A J, O'Brien C N, Nickerson S C, Marquardt W W

机构信息

Milk Secretion and Mastitis Laboratory, Agricultural Research Service, USDA, Beltsville, MD 20705.

出版信息

J Dairy Sci. 1994 Apr;77(4):970-83. doi: 10.3168/jds.S0022-0302(94)77033-8.

Abstract

Bovine mammary secretory cells, isolated at necropsy, were cultured in vitro and used as a model to study the mode of adherence of Staphylococcus aureus to mammary epithelium. Cultured cells were characterized by their morphology and physiology as secretory epithelial cells. Cells showed characteristic growth patterns when grown on polystyrene, fibronectin, laminin, collagen, and reconstituted basement membrane from the Engelbreth-Holm-Swarm murine sarcoma. Cells cultured on collagen formed confluent monolayers and were the most suitable for bacterial adherence studies. Cultured cells stained intensely for cytokeratin and for specific milk proteins, i.e., alpha-casein, beta-casein, alpha-lactalbumin, beta-lactoglobulin, and lactoferrin. The effect of frozen storage for 10 mo on cell viability or presence of milk proteins was minimal. Staphylococcus aureus showed large affinity for extracellular matrix components, i.e., fibronectin, laminin, and collagen. Adherence to confluent cell monolayers was minimal. In preconfluent cell monolayers, most S. aureus adhered more readily to the exposed matrix than to the epithelial cells. Overnight exposure to staphylococcal alpha-toxin greatly increased adherence of S. aureus to confluent monolayers. However, whether bacteria adhered to alpha-toxin damaged cells or to exposed matrix is not clear. Unencapsulated S. aureus adhered in larger numbers than did encapsulated S. aureus.

摘要

在尸检时分离出的牛乳腺分泌细胞在体外进行培养,并用作研究金黄色葡萄球菌黏附于乳腺上皮细胞模式的模型。培养的细胞通过其形态和生理学特征被鉴定为分泌上皮细胞。当这些细胞在聚苯乙烯、纤连蛋白、层粘连蛋白、胶原蛋白以及来自Engelbreth-Holm-Swarm小鼠肉瘤的重组基底膜上生长时,呈现出特征性的生长模式。在胶原蛋白上培养的细胞形成汇合的单层,最适合用于细菌黏附研究。培养的细胞对细胞角蛋白和特定的乳蛋白,即α-酪蛋白、β-酪蛋白、α-乳白蛋白、β-乳球蛋白和乳铁蛋白,染色强烈。冷冻保存10个月对细胞活力或乳蛋白的存在影响极小。金黄色葡萄球菌对细胞外基质成分,即纤连蛋白、层粘连蛋白和胶原蛋白,表现出很大的亲和力。对汇合的细胞单层的黏附极少。在未汇合的细胞单层中,大多数金黄色葡萄球菌更容易黏附于暴露的基质而非上皮细胞。过夜暴露于葡萄球菌α-毒素可大大增加金黄色葡萄球菌对汇合单层的黏附。然而,细菌是黏附于α-毒素损伤的细胞还是暴露的基质尚不清楚。未包膜的金黄色葡萄球菌比包膜的金黄色葡萄球菌黏附数量更多。

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