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Uropathogenic Escherichia coli Flagella Aid in Efficient Urinary Tract Colonization

机译：尿毒症大肠杆菌鞭毛有助于有效尿路定植

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In the murine model of urinary tract infections (UTI), cystitis by uropathogenic Escherichia coli (UPEC) occurs through an intimate relationship with the bladder superficial umbrella cell entailing cycles of adherence, invasion, intracellular bacterial community (IBC) formation, and dispersal (fluxing) from the intracellular environment. IBC dispersal is a key step that results in the spread of bacteria over the epithelial surface to initiate additional rounds of IBC formation. We investigated the role of flagella in mediating adherence and motility during UTI, hypothesizing that the dispersion of the IBC would be incomplete in the absence of motility, thus interrupting the IBC pathway and attenuating the infection. Using gfp reporter fusions, the expression of the flagellar class I flhDC and class III fliC genes was monitored to track key points of regulation throughout the pathogenic cascade. In vitro, growth under conditions promoting motility resulted in the robust expression of both fusions. In contrast, only the class I fusion produced significant expression throughout early stages of IBC development including the dispersion stage. Thus, unlike in vitro modeling of motility, the regulatory cascade appeared incomplete in vivo. Throughout IBC formation, nonmotile ΔfliC mutants achieved the same number of IBCs as the wild-type (wt) strain, demonstrating that flagella are neither essential nor required for first- or second-generation IBC formation. However, in competition experiments between wt and ΔfliC strains, the wt was shown to have a fitness advantage in persisting throughout the urinary tract for 2 weeks, demonstrating a subtle but measurable role for flagella in virulence.
机译：在尿路感染（UTI）的鼠模型中，尿路致病性大肠埃希氏菌（UPEC）引起的膀胱炎是通过与膀胱浅层伞形细胞的紧密关系发生的，从而导致粘附，侵袭和细胞内细菌群落（IBC） ）形成，并从细胞内环境扩散（助焊剂）。 IBC扩散是关键步骤，可导致细菌在上皮表面扩散，从而引发新一轮的IBC形成。我们调查了鞭毛在UTI期间介导粘附和运动的作用，假设在没有运动的情况下IBC的分散是不完全的，从而中断了IBC途径并减弱了感染。使用 gfp 报告基因融合体，监测鞭毛I类 flhDC 和III类 fliC 基因的表达，以追踪整个致病性调控的关键点级联。在体外，在促进运动的条件下生长导致两种融合物的稳健表达。相比之下，只有I类融合蛋白在IBC发展的整个早期阶段（包括分散阶段）都产生了重要的表达。因此，与体外运动模型不同，调节级联在体内似乎是不完全的。在整个IBC形成过程中，非活动性Δ fliC 突变体实现了与野生型（ wt ）菌株相同的IBC数量，这表明鞭毛既不是初生也不是必需的第二代IBC的形成。然而，在 wt 和Δ fliC 菌株之间的竞争实验中， wt 被证明具有在整个尿道中坚持2次的健身优势。数周，证明鞭毛在毒力中具有微妙但可测量的作用。

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