Department of Biochemistry and Medical Biotechnology, School of Tropical Medicine, 108, C.R. Avenue, Kolkata, 700073, India.
School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW, Australia.
Folia Microbiol (Praha). 2024 Dec;69(6):1185-1204. doi: 10.1007/s12223-024-01159-y. Epub 2024 Apr 3.
The adherence of bladder uroepithelial cells, subsequent expression, and regulation of type 1 fimbrial genes (key mediator of attachment) in clinical multidrug-resistant uropathogenic Escherichia coli (MDR-UPECs) isolated from individuals with asymptomatic bacteriuria (ABU) remain unexplored till date. Therefore, this study aimed to investigate the underlying molecular mechanisms associated with the adherence of clinical MDR-ABU-UPECs to human a uroepithelial cell line (HTB-4), both in the absence and presence of D-Mannose. These investigations focused on phase variation, expression, and regulation of type 1 fimbriae and were compared to a prototype ABU-strain (E. coli 83972) and symptomatic MDR-UPECs. Discordant to the ABU prototype strain, MDR-ABU-UPECs exhibited remarkable adhesive capacity that was significantly reduced after D-mannose exposure, fairly like the MDR symptomatic UPECs. The type 1 fimbrial phase variation, determined by the fim switch analysis, asserted the statistically significant incidence of "both OFF and ON" orientation among the adherent MDR-ABU-UPECs with a significant reduction in phase-ON colonies post-D-mannose exposure, akin to the symptomatic ones. This was indicative of an operative and alternating type 1 fimbrial phase switch. The q-PCR assay revealed a coordinated action of the regulatory factors; H-NS, IHF, and Lrp on the expression of FimB and FimE recombinases, which further controlled the function of fimH and fimA genes in ABU-UPECs, similar to symptomatic strains. Therefore, this study is the first of its kind to provide an insight into the regulatory crosstalk of different cellular factors guiding the adhesion of ABU-UPECs to the host. Additionally, it also advocated for the need to accurately characterize ABU-UPECs.
迄今为止,尚未探索从无症状性菌尿(ABU)个体中分离出的临床多药耐药尿路致病性大肠埃希菌(MDR-UPEC)中膀胱尿路上皮细胞的黏附、随后的 1 型菌毛基因(黏附的关键介质)的表达和调控。因此,本研究旨在研究与临床 MDR-ABU-UPEC 与人尿路上皮细胞系(HTB-4)黏附相关的潜在分子机制,包括在缺乏和存在 D-甘露糖的情况下。这些研究集中在 1 型菌毛的表型变异、表达和调控上,并与原型 ABU 株(E. coli 83972)和有症状的 MDR-UPEC 进行了比较。与 ABU 原型株不同的是,MDR-ABU-UPEC 表现出显著的黏附能力,在 D-甘露糖暴露后显著降低,与 MDR 有症状 UPEC 相当。通过 fim 开关分析确定的 1 型菌毛表型变异表明,在黏附的 MDR-ABU-UPEC 中,“关”和“开”两种取向的发生率显著,在 D-甘露糖暴露后,“开”取向的菌落显著减少,类似于有症状的菌株。这表明存在一种有效的交替 1 型菌毛表型开关。q-PCR 检测显示,调节因子 H-NS、IHF 和 Lrp 对 FimB 和 FimE 重组酶的表达具有协同作用,这进一步控制了 ABU-UPEC 中 fimH 和 fimA 基因的功能,类似于有症状的菌株。因此,这项研究首次提供了对不同细胞因子调控 ABU-UPEC 与宿主黏附的调控对话的深入了解。此外,它还强调了需要准确地对 ABU-UPEC 进行特征描述。