Biomimetic Materials to Characterize Bacteria-host Interactions.
Biomimetic Materials to Characterize Bacteria-host Interactions.
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DOI：
10.3791/53400
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发表时间：
2015-11-16
期刊：
影响因子：
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通讯作者：
Krachler AM
中科院分区：
文献类型：
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作者：
Stones DH;Al-Saedi F;Vaz D;Perez-Soto N;Krachler AM
细菌对宿主细胞的附着是细菌在宿主组织定殖过程中最早的事件之一，因此是感染过程中的关键步骤。介导这些初始细菌-宿主相互作用的粘附素的生物化学和功能表征通常受到干扰分析的其他细菌因子（例如细胞壁组分或分泌分子）的存在的损害。该方案描述了仿生材料的生产和使用，由纯的重组粘附素化学偶联到市售的，功能化的聚苯乙烯珠，已成功地用于解剖单个细菌粘附素和宿主细胞受体之间的生物化学和功能相互作用。协议不同的耦合化学，允许定向固定的重组粘附素聚合物支架上，并评估所产生的“拟细菌”材料的耦合效率进行了讨论。我们进一步描述了如何将这些材料用作抑制病原体介导的细胞毒性的工具，并讨论了这种方法在研究细菌-宿主相互作用中的范围、限制和进一步应用。
Bacterial attachment to host cells is one of the earliest events during bacterial colonization of host tissues and thus a key step during infection. The biochemical and functional characterization of adhesins mediating these initial bacteria-host interactions is often compromised by the presence of other bacterial factors, such as cell wall components or secreted molecules, which interfere with the analysis. This protocol describes the production and use of biomimetic materials, consisting of pure recombinant adhesins chemically coupled to commercially available, functionalized polystyrene beads, which have been used successfully to dissect the biochemical and functional interactions between individual bacterial adhesins and host cell receptors. Protocols for different coupling chemistries, allowing directional immobilization of recombinant adhesins on polymer scaffolds, and for assessment of the coupling efficiency of the resulting “bacteriomimetic” materials are also discussed. We further describe how these materials can be used as a tool to inhibit pathogen mediated cytotoxicity and discuss scope, limitations and further applications of this approach in studying bacterial - host interactions.