700928 cft073 none wam 2267  (ATCC)

Journal: Infection and Immunity

Article Title: Human body temperature cues widespread changes in virulence gene expression in uropathogenic Escherichia coli

doi: 10.1128/iai.00422-25

Figure Lengend Snippet: Characterization and comparison of the transcriptome and proteome of uropathogen E. coli CFT073 4 h after a shift from 23°C to 37°C. ( A ) Comparison of the differentially regulated genes and proteins 4 h after the temperature upshift from 23°C to 37°C. ( B ) Volcano plots of the transcriptome and proteome with each dot representing a unique mRNA/protein. mRNAs/proteins with significantly increased expression (magenta) or decreased expression (green) (FDR ≤ 1%, at least ±1 log2FC) are indicated. Volcano plots were created using VolcaNoseR . ( C ) For genes detected in both the transcriptome and proteome, the 37°C/23°C log2FC of each mRNA is plotted on the x -axis in comparison to its 37°C/23°C log2FC protein on the y -axis. ( D ) Comparison of 37°C/23°C log2FC for mRNA (blue dot) to protein (orange dot) for those genes with FDR ≤ 1% and at least ±1 log2FC in both the transcriptome and proteome. ( E ) Comparison of differentially regulated genes in E. coli CFT073 to genes in E. coli MC4100.

Article Snippet: The strains used in this study include CFT073 ( ) and CFT073 rpoS (WAM 2267/ATCC 700928) ( ).

Techniques: Comparison, Expressing

Journal: Infection and Immunity

Article Title: Human body temperature cues widespread changes in virulence gene expression in uropathogenic Escherichia coli

doi: 10.1128/iai.00422-25

Figure Lengend Snippet: Impacts of a shift to 37°C on RpoS protein levels, regulators of RpoS expression, and RpoS-regulated genes. ( A ) Western blotting of RpoS protein in cultures retained at 23°C and those shifted to 37°C. + control = purified RpoS protein, − control = total protein from ∆ rpoS strain. Alkaline phosphatase (AP) was used as a non-thermoregulated control. A representative experiment is shown. ( B ) 37°C/23°C log2FC mRNA of rpoS and the regulators of RpoS stability ( iraL ) and RpoS translation (DsrA, RprA). ( C ) Analysis of CFT073 thermoregulated genes that are RpoS-dependent in E. coli K-12 MC4100.

Article Snippet: The strains used in this study include CFT073 ( ) and CFT073 rpoS (WAM 2267/ATCC 700928) ( ).

Techniques: Expressing, Western Blot, Control, Purification

Journal: Infection and Immunity

Article Title: Human body temperature cues widespread changes in virulence gene expression in uropathogenic Escherichia coli

doi: 10.1128/iai.00422-25

Figure Lengend Snippet: Diagram emphasizing the temperature-regulated genes in uropathogenic E. coli CFT073 that show differential expression after a temperature upshift from 23°C to 37°C. They are grouped by function including adhesion and biofilm formation (black), immune evasion (purple), and competitor defense (maroon). Created in https://BioRender.com .

Article Snippet: The strains used in this study include CFT073 ( ) and CFT073 rpoS (WAM 2267/ATCC 700928) ( ).

Techniques: Quantitative Proteomics

Journal: Infection and Immunity

Article Title: Human body temperature cues widespread changes in virulence gene expression in uropathogenic Escherichia coli

doi: 10.1128/iai.00422-25

Figure Lengend Snippet: Characterization and comparison of the transcriptome and proteome of uropathogen E. coli CFT073 4 h after a shift from 23°C to 37°C. ( A ) Comparison of the differentially regulated genes and proteins 4 h after the temperature upshift from 23°C to 37°C. ( B ) Volcano plots of the transcriptome and proteome with each dot representing a unique mRNA/protein. mRNAs/proteins with significantly increased expression (magenta) or decreased expression (green) (FDR ≤ 1%, at least ±1 log2FC) are indicated. Volcano plots were created using VolcaNoseR . ( C ) For genes detected in both the transcriptome and proteome, the 37°C/23°C log2FC of each mRNA is plotted on the x -axis in comparison to its 37°C/23°C log2FC protein on the y -axis. ( D ) Comparison of 37°C/23°C log2FC for mRNA (blue dot) to protein (orange dot) for those genes with FDR ≤ 1% and at least ±1 log2FC in both the transcriptome and proteome. ( E ) Comparison of differentially regulated genes in E. coli CFT073 to genes in E. coli MC4100.

Article Snippet: The strains used in this study include CFT073 ( ) and CFT073 rpoS (WAM 2267/ATCC 700928) ( ).

Techniques: Comparison, Expressing

Journal: Infection and Immunity

Article Title: Human body temperature cues widespread changes in virulence gene expression in uropathogenic Escherichia coli

doi: 10.1128/iai.00422-25

Figure Lengend Snippet: Impacts of a shift to 37°C on RpoS protein levels, regulators of RpoS expression, and RpoS-regulated genes. ( A ) Western blotting of RpoS protein in cultures retained at 23°C and those shifted to 37°C. + control = purified RpoS protein, − control = total protein from ∆ rpoS strain. Alkaline phosphatase (AP) was used as a non-thermoregulated control. A representative experiment is shown. ( B ) 37°C/23°C log2FC mRNA of rpoS and the regulators of RpoS stability ( iraL ) and RpoS translation (DsrA, RprA). ( C ) Analysis of CFT073 thermoregulated genes that are RpoS-dependent in E. coli K-12 MC4100.

Article Snippet: The strains used in this study include CFT073 ( ) and CFT073 rpoS (WAM 2267/ATCC 700928) ( ).

Techniques: Expressing, Western Blot, Control, Purification

Journal: Infection and Immunity

Article Title: Human body temperature cues widespread changes in virulence gene expression in uropathogenic Escherichia coli

doi: 10.1128/iai.00422-25

Figure Lengend Snippet: Diagram emphasizing the temperature-regulated genes in uropathogenic E. coli CFT073 that show differential expression after a temperature upshift from 23°C to 37°C. They are grouped by function including adhesion and biofilm formation (black), immune evasion (purple), and competitor defense (maroon). Created in https://BioRender.com .

Article Snippet: The strains used in this study include CFT073 ( ) and CFT073 rpoS (WAM 2267/ATCC 700928) ( ).

Techniques: Quantitative Proteomics

Journal: Biomedicines

Article Title: Trained Immunity in Bladder ILC3s Enhances Mucosal Defense Against Recurrent Urinary Tract Infections

doi: 10.3390/biomedicines14010078

Figure Lengend Snippet: ILC3s increase systematically during UPEC-induced urinary tract infection. ( a ) Representative immunofluorescence staining and quantification of ILC3s (CD3−RORγt + cells, white asterisk (*)) in human bladder sections from healthy controls (HC, n = 10) and UTI patients ( n = 9). ( b ) Schematic of the mouse acute UTI model. C57BL/6 mice were transurethrally inoculated with 5 × 10 7 CFU UPEC strain CFT073 or PBS and analyzed 24 h post-infection. ( c ) Representative images of bacterial cultures from bladder and kidney homogenates. ( d , e ) Representative flow cytometry plots and quantification of ILC3s in mouse bladder ( d ) and kidney ( e ). ( f ) Flow cytometry analysis of IL-17A and IL-22 expression by bladder ILC3s. Data are shown as mean ± SEM. * p < 0.05, **** p < 0.0001, ns p > 0.05.

Article Snippet: Uropathogenic Escherichia coli ( E. coli ) strain CFT073 (ATCC 700928, isolated from an acute pyelonephritis patient’s clinical specimen) was used.

Techniques: Infection, Immunofluorescence, Staining, Flow Cytometry, Expressing

Journal: Biomedicines

Article Title: Trained Immunity in Bladder ILC3s Enhances Mucosal Defense Against Recurrent Urinary Tract Infections

doi: 10.3390/biomedicines14010078

Figure Lengend Snippet: ILC3s contribute to protection against UTI. ( a ) Schematic of the adoptive transfer and infection model. ( b – e ) Validation of ILC3 transfer: ILC3 counts ( b , c ) and representative flow cytometry plots ( d , e ) in bladder and kidney 24 h after UPEC challenge of mice received ILC3s or PBS transfer. ( f ) Schematic of experimental design. ( g ) Flow cytometry analysis of donor (CD45.1 + ) versus host (CD45.2 + ) ILC3s in recipient bladders. The orange polygon denotes CD45.1 + ILC3s, the black polygon denotes CD45.2 + white blood cells (WBCs), the black circle denotes CD45.2 + ILCs, and the blue polygon denotes CD45.2 + ILC3s. ( h , i ) Disease severity assessed by bladder/body weight ratio ( h ) and body weight change ( i ). ( j , k ) Bacterial load of bladders ( j ) and kidneys ( k ). ( l , m ) Representative immunofluorescence images ( l ) and mean fluorescence intensity (MFI) quantification ( m ) of UPEC (green) in bladder mucosa. Scale bars = 50 µm. ( n – p ) Bladder histopathology was assessed on H&E-stained sections ( n ), scale bars = 50 µm, with quantification of submucosal edema ( o ) and pathological scores ( p ). Data are shown as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns p > 0.05.

Article Snippet: Uropathogenic Escherichia coli ( E. coli ) strain CFT073 (ATCC 700928, isolated from an acute pyelonephritis patient’s clinical specimen) was used.

Techniques: Adoptive Transfer Assay, Infection, Biomarker Discovery, Flow Cytometry, Immunofluorescence, Fluorescence, Histopathology, Staining

Journal: Biomedicines

Article Title: Trained Immunity in Bladder ILC3s Enhances Mucosal Defense Against Recurrent Urinary Tract Infections

doi: 10.3390/biomedicines14010078

Figure Lengend Snippet: UPEC-trained ILC3s confer enhanced protection against recurrent UTI through pathogen clearance and tissue preservation. ( a ) Schematic of experimental design for primary and secondary infection. ( b , c ) Body weight change ( b ) and bladder ILC3 counts ( c ) monitored over the infection timeline. ( d – f ) ILC, ILC3 counts and the proportion of ILC3s among ILCs in bladder were detected by flow cytometry analysis. ( g ) Schematic of experimental design for adoptive transfer of naïve or Tr-ILC3s. ( h , i ) Disease severity in recipient mice assessed by bladder/body weight ratio ( h ) and body weight change ( i ) at 24 h post-infection. ( j – l ) Bacterial burden was assessed with representative culture plates ( j ) and quantified CFU in bladder ( k ) and kidney ( l ). ( m ) Representative images of H&E-stained bladder sections. Scale bars = 50 μm. ( n – p ) The percentage of subepidermal lamina propria edema in the urothelium ( n ), pathological scores ( o ), and the percentage of epithelial cell sloughing ( p ) were analyzed based on H&E-stained bladder sections. Data are shown as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, ns p > 0.05.

Article Snippet: Uropathogenic Escherichia coli ( E. coli ) strain CFT073 (ATCC 700928, isolated from an acute pyelonephritis patient’s clinical specimen) was used.

Techniques: Preserving, Infection, Flow Cytometry, Adoptive Transfer Assay, Staining

Journal: Biomedicines

Article Title: Trained Immunity in Bladder ILC3s Enhances Mucosal Defense Against Recurrent Urinary Tract Infections

doi: 10.3390/biomedicines14010078

Figure Lengend Snippet: Tr-ILC3s acquire cell-intrinsic functional and proliferative advantages. ( a ) Uroplakin IIIa expression in bladder sections assessed by immunofluorescence staining (green) and MFI quantification. ( b , c ) The mRNA expression of Reg3b , Reg3g , S100a8 and S100a9 in bladders ( b ) and kidneys ( c ) was assessed by real-time qPCR. ( d – f ) The mRNA expression of Il6 , Tnfa and Ifng in bladders was assessed by real-time qPCR. ( g ) ILC3 counts in bladder following transfer of naïve or Tr-ILC3s. ( h ) Proliferation of bladder ILC3s assessed by flow cytometry for Ki67 expression. ( i ) IL-17A and IL-22 expression on ILC3s in bladder were detected by flow cytometry. ( j ) IL-17A and IL-22 expression by MNK-3 cells in vitro after primary or secondary stimulation with UPEC lysate, analyzed by flow cytometry. Data are shown as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns p > 0.05.

Article Snippet: Uropathogenic Escherichia coli ( E. coli ) strain CFT073 (ATCC 700928, isolated from an acute pyelonephritis patient’s clinical specimen) was used.

Techniques: Functional Assay, Expressing, Immunofluorescence, Staining, Flow Cytometry, In Vitro