Phosphoethanolamine cellulose enhances curli-mediated adhesion of uropathogenic Escherichia coli to bladder epithelial cells
Phosphoethanolamine cellulose enhances curli-mediated adhesion of uropathogenic Escherichia coli to bladder epithelial cells
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DOI：
10.1073/pnas.1801564115
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发表时间：
2018-10-02
影响因子：
11.1
通讯作者：
Cegelski, Lynette
中科院分区：
文献类型：
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作者：
Hollenbeck, Emily C.;Antonoplis, Alexandra;Cegelski, Lynette
尿路致病性大肠杆菌（UPEC）是尿路感染的主要病原体，采用许多分子策略，以促进粘附，定植和持久性在膀胱龛。确定防止粘附和定殖的策略是抑制细菌发病机制和帮助保持可用抗生素功效的有希望的方法。这种方法需要更好地了解粘附到膀胱尿路上皮的分子决定因素。我们设计的实验，使用定制的活细胞单层流变仪（LCMR）定量测量单个和合并的贡献细菌细胞表面结构[1型皮利，卷曲，磷酸乙醇胺（pEtN）纤维素]膀胱细胞粘附。使用UPEC菌株UTI89、同基因突变体和用于细胞表面结构的差异生产的受控条件，我们发现卷曲可以促进比1型皮利更强的与膀胱细胞的粘附相互作用。此外，curli和pEtN纤维素的共同生产增强了粘附性。LCMR能够在高剪切条件下评价粘附性，以揭示pEtN纤维素的这种作用，而pEtN纤维素使用常规组织培养粘附测定法无法检测到。连同补充的生物化学实验，结果支持一个模型，其中纤维素起到砂浆样的功能，以促进卷曲协会与周围的细菌细胞表面，导致在膀胱细胞表面的细菌粘附强度增加。
Uropathogenic Escherichia coli (UPEC) are the major causative agents of urinary tract infections, employing numerous molecular strategies to contribute to adhesion, colonization, and persistence in the bladder niche. Identifying strategies to prevent adhesion and colonization is a promising approach to inhibit bacterial pathogenesis and to help preserve the efficacy of available antibiotics. This approach requires an improved understanding of the molecular determinants of adhesion to the bladder urothelium. We designed experiments using a custom-built live cell monolayer rheometer (LCMR) to quantitatively measure individual and combined contributions of bacterial cell surface structures [type 1 pili, curli, and phosphoethanolamine (pEtN) cellulose] to bladder cell adhesion. Using the UPEC strain UTI89, isogenic mutants, and controlled conditions for the differential production of cell surface structures, we discovered that curli can promote stronger adhesive interactions with bladder cells than type 1 pili. Moreover, the coproduction of curli and pEtN cellulose enhanced adhesion. The LCMR enables the evaluation of adhesion under high-shear conditions to reveal this role for pEtN cellulose which escaped detection using conventional tissue culture adhesion assays. Together with complementary biochemical experiments, the results support a model wherein cellulose serves a mortar-like function to promote curli association with and around the bacterial cell surface, resulting in increased bacterial adhesion strength at the bladder cell surface.