Wyatt J E, Poston S M, Noble W C
Department of Microbiology, United Medical School of Guy Hospital, London, UK.
J Appl Bacteriol. 1990 Dec;69(6):834-44. doi: 10.1111/j.1365-2672.1990.tb01581.x.
Adherence of four strains of Staphylococcus aureus to eukaryotic cell monolayers was assayed with [3H]-thymidine labelled bacterial cells and the results were analysed by non-parametric statistical tests. Adherence to primary (human mesothelial) and semi-continuous (human embryonic lung) cell monolayers was significantly better than to continuous cell lines (HEp2, HeLa and Vero). HEp2 cell monolayers provided the most reliable assay substrate of the continuous cell lines tested. Variation occurred between bacterial culture batches but the assay measured significant differences between adhesion levels of the strains and distinguished between high level (RN92, 8325-4) and low level (Wood46, ISP458) adhering strains. Adherence to different batches of cell monolayers also varied but relative adherence values for strains were similar and the ranking of strains according to adhesion values was unchanged. Potential adhesion mediators have been monitored for their effect on adhesion of a highly adherent strain (RN92) to HEp2 monolayers. Fibronectin, protein A and anti-protein A did not significantly affect adhesion. Lipoteichoic acid caused a significant inhibition of adhesion. With critical statistical analysis to accommodate inherent variations, this assay provides a useful model to study factors involved in adherence of Staph. aureus to eukaryotic cells.
用[3H] - 胸腺嘧啶核苷标记的细菌细胞检测了四株金黄色葡萄球菌对真核细胞单层的黏附情况,并通过非参数统计检验分析了结果。与连续细胞系(HEp2、HeLa和Vero)相比,对原代(人间皮)和半连续(人胚肺)细胞单层的黏附明显更好。在测试的连续细胞系中,HEp2细胞单层提供了最可靠的检测底物。不同细菌培养批次之间存在差异,但该检测方法能够检测出菌株黏附水平的显著差异,并区分高黏附水平菌株(RN92、8325 - 4)和低黏附水平菌株(Wood46、ISP458)。对不同批次细胞单层的黏附情况也有所不同,但菌株的相对黏附值相似,根据黏附值对菌株的排名没有变化。已监测潜在黏附介质对高黏附菌株(RN92)黏附到HEp2单层的影响。纤连蛋白、蛋白A和抗蛋白A对黏附没有显著影响。脂磷壁酸对黏附产生了显著抑制作用。通过关键的统计分析以适应内在差异,该检测方法为研究金黄色葡萄球菌黏附真核细胞的相关因素提供了一个有用的模型。