Differential colony size, cell length, and cellular proteome of Escherichia coli isolated from urine vs. stone nidus of kidney stone patients

Tavichakorntrakool, Ratree; Boonsiri, Patcharee; Prasongwatana, Vitoon; Lulitanond, Aroonlug; Wongkham, Chaisiri; Thongboonkerd, Visith

doi:10.1016/j.cca.2016.12.018

Cited by 22 publications

(16 citation statements)

References 16 publications

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“…Phenotypic characteristics, including bacterial colony size, cell length and time to mid-log phase of the growth curve, which may be the important factors for adaptive response of bacteria to survive within different environments (e.g., inside vs. outside the stone matrix) 10 , were examined. The findings showed no significant differences of these physical characteristics in EUU vs. EUK groups (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Elongation factor Tu on Escherichia coli isolated from urine of kidney stone patients promotes calcium oxalate crystal growth and aggregation

Amimanan

Tavichakorntrakool

Fong-ngern

et al. 2017

Sci Rep

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Escherichia coli is the most common bacterium isolated from urine and stone matrix of calcium oxalate (CaOx) stone formers. Whether it has pathogenic role(s) in kidney stone formation or is only entrapped inside the stone remains unclear. We thus evaluated differences between E. coli isolated from urine of patients with kidney stone (EUK) and that from patients with urinary tract infection (UTI) without stone (EUU). From 100 stone formers and 200 UTI patients, only four pairs of EUK/EUU isolates had identical antimicrobial susceptibility patterns. Proteomic analysis revealed nine common differentially expressed proteins. Among these, the greater level of elongation factor Tu (EF-Tu) in EUK was validated by Western blotting. Outer membrane vesicles (OMVs) derived from EUK had greater promoting activities on CaOx crystallization, crystal growth and aggregation as compared to those derived from EUU. Neutralizing the OMVs of EUK with monoclonal anti-EF-Tu antibody, not with an isotype antibody, significantly reduced all these OMVs-induced promoting effects. Moreover, immunofluorescence staining of EF-Tu on bacterial cell surface confirmed the greater expression of surface EF-Tu on EUK (vs. EUU). Our data indicate that surface EF-Tu and OMVs play significant roles in promoting activities of E. coli on CaOx crystallization, crystal growth and aggregation.

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Section: Resultsmentioning

confidence: 99%

Elongation factor Tu on Escherichia coli isolated from urine of kidney stone patients promotes calcium oxalate crystal growth and aggregation

Amimanan

Tavichakorntrakool

Fong-ngern

et al. 2017

Sci Rep

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“…More recent studies have provided increasing evidence of the possible roles of non-urease-producing bacteria in kidney stone formation in addition to their roles in the secondary infection following urological obstruction or urinary stagnation from renal calculi (Barr-Beare et al, 2015; Tavichakorntrakool et al, 2017). The study on interaction between Enterobacteriaceae and deposition of CaOx stone in pediatric urolithiasis showed that E. coli was identified in a positive stone culture and selectively aggregated on and around CaOx crystals (Barr-Beare et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Flagellum Is Responsible for Promoting Effects of Viable Escherichia coli on Calcium Oxalate Crystallization, Crystal Growth, and Crystal Aggregation

2019

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Urease-producing bacteria (especially Proteus mirabilis) can cause infection kidney stone. However, recent studies have shown that intact viable non-urease-producing bacteria such as Escherichia coli might also promote calcium oxalate (CaOx) kidney stone formation but with unclear mechanism. We thus hypothesized that some relevant bacterial components might be responsible for such promoting effects of the intact viable E. coli. Flagella, capsule, lipopolysaccharide (LPS), and outer membrane vesicles (OMVs) were isolated/purified and their stone modulatory activities were evaluated using CaOx crystallization, crystal growth, and crystal aggregation assays. Among these, flagella had the most potent promoting effects on CaOx crystallization, crystal growth, and crystal aggregation. Validation was performed by deflagellation demonstrating that the deflagellated intact viable E. coli had markedly reduced CaOx crystal modulatory activities in all aspects (comparable to those of the negative controls). Similarly, neutralization of the isolated/purified flagella using a specific anti-flagellin antibody, not an isotype control, could abolish the promoting effects of flagella. These findings provide direct evidence indicating that flagellum is responsible for the promoting effects of the viable E. coli on CaOx crystallization, crystal growth and aggregation.

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“…Separation of the digested peptides was performed using EASY-nLC II (Bruker Daltonics, Bremen, Germany) as previously described ( 26 , 27 ). Briefly, peptides were loaded from a cooled (7°C) autosampler into an in-house, 3-cm-long pre-column containing 5-µm C18 resin (Dr. Maisch GmbH, Ammerbuch, Germany) and then to an in-house, 10-cm-long analytical column packed with 3-µm C18 resin (Dr. Maisch GmbH) using mobile phase A (0.1% formic acid).…”

Section: Methodsmentioning

confidence: 99%

Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals

et al. 2018

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In kidney stone disease, macrophages secrete various mediators via classical secretory pathway and cause renal interstitial inflammation. However, whether their extracellular vesicles, particularly exosomes, are involved in kidney stone pathogenesis remained unknown. This study investigated alterations in exosomal proteome of U937-derived macrophages (by phorbol-12-myristate-13-acetate activation) after exposure to calcium oxalate monohydrate (COM) crystals for 16-h using 2-DE-based proteomics approach. Six significantly altered proteins in COM-treated exosomes were successfully identified by nanoscale liquid chromatography–electrospray ionization–electron transfer dissociation tandem mass spectrometry as proteins involved mainly in immune processes, including T-cell activation and homeostasis, Fcγ receptor-mediated phagocytosis, interferon-γ (IFN-γ) regulation, and cell migration/movement. The decreased heat shock protein 90-beta (HSP90β) and increased vimentin were confirmed by Western blotting. ELISA showed that the COM-treated macrophages produced greater level of interleukin-1β (IL-1β), one of the markers for inflammasome activation. Functional studies demonstrated that COM-treated exosomes enhanced monocyte and T-cell migration, monocyte activation and macrophage phagocytic activity, but on the other hand, reduced T-cell activation. In addition, COM-treated exosomes enhanced production of proinflammatory cytokine IL-8 by monocytes that could be restored to its basal level by small-interfering RNA targeting on vimentin (si-Vimentin). Moreover, si-Vimentin could also abolish effects of COM-treated exosomes on monocyte and T-cell migration as well as macrophage phagocytic activity. These findings provided some implications to the immune response during kidney stone pathogenesis via exosomal pathway of macrophages after exposure to COM crystals.

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Differential colony size, cell length, and cellular proteome of Escherichia coli isolated from urine vs. stone nidus of kidney stone patients

Cited by 22 publications

References 16 publications

Elongation factor Tu on Escherichia coli isolated from urine of kidney stone patients promotes calcium oxalate crystal growth and aggregation

Elongation factor Tu on Escherichia coli isolated from urine of kidney stone patients promotes calcium oxalate crystal growth and aggregation

Flagellum Is Responsible for Promoting Effects of Viable Escherichia coli on Calcium Oxalate Crystallization, Crystal Growth, and Crystal Aggregation

Roles of Macrophage Exosomes in Immune Response to Calcium Oxalate Monohydrate Crystals

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