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Analysis of the dynamics of Staphylococcus aureus binding to white blood cells using whole blood assay and geno-to-pheno mapping.
International Journal of Medical Microbiology ( IF 4.1 ) Pub Date : 2020-02-05 , DOI: 10.1016/j.ijmm.2020.151411
Daria Gaidar 1 , Alice Jonas 2 , Ruslan Akulenko 1 , Ulla Ruffing 2 , Mathias Herrmann 2 , Volkhard Helms 3 , Lutz von Müller 2
Affiliation  

Given that binding and internalization of bacteria to host cells promotes infections and invasion, we aimed at characterizing how various S. aureus isolates adhere to and are internalized by different white blood cells. In particular, the role of genetic determinants on the association kinetics should be unveiled. A flow cytometric (FACS) whole blood assay with fluorescently labelled isolates was applied to 56 clinical S. aureus isolates. This phenotypic data was then linked to previously obtained genotyping data (334 genes) with the help of a redescription mining algorithm. Professional phagocytes showed a time-dependent increase of bacterial adhesion and internalization. Isolates showing higher affinity to granulocytes were associated with lower binding to monocytes. In contrast binding activity between S. aureus and lymphocytes could be subdivided into two phases. Preliminary binding (phase 1) was highest directly after co-incubation and was followed by S. aureus detachment or by sustained binding of a small lymphocyte subset (phase 2). Strain-dependent low granulocyte binding was observed for clonal complex 5 (CC5) isolates (MRSA), as compared to CC30 and CC45 (MSSA). S. aureus isolates associated with low granulocyte phagocytosis were characterized by the presence (cap8, can) and the absence (sasG, lukD, isdA, splA, setC) of specific hybridization signals.



中文翻译:

金黄色葡萄球菌与白细胞结合的动力学分析,采用全血分析和基因-表型作图。

鉴于细菌与宿主细胞的结合和内在化促进了感染和入侵,我们旨在表征各种金黄色葡萄球菌分离物如何粘附并被不同的白细胞内化。特别是,应该揭示遗传决定因素在缔合动力学上的作用。将流式细胞术(FACS)全血测定与荧光标记的分离物应用于56株临床金黄色葡萄球菌隔离株。然后,借助重新定义挖掘算法,将该表型数据与先前获得的基因分型数据(334个基因)相关联。专业吞噬细胞显示细菌粘附和内在化的时间依赖性增加。对粒细胞显示出更高亲和力的分离物与对单核细胞的结合降低。相反,金黄色葡萄球菌和淋巴细胞之间的结合活性可分为两个阶段。共孵育后,初步结合(阶段1)最高,随后是金黄色葡萄球菌脱离或小淋巴细胞亚群的持续结合(阶段2)。与CC30和CC45(MSSA)相比,克隆复合物5(CC5)分离株(MRSA)观察到了应变依赖性的低粒细胞结合。金黄色葡萄球菌与低粒细胞吞噬作用相关的分离株的特征是存在(cap8罐头)和不存在(sasGlukDisdAsplAsetC)特异性杂交信号。

更新日期:2020-02-05
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