Mean spectra projection (MSP) dendrogram of <i>Klebsiella pneumoniae</i> strains generated by BIOTYPER software (version 2; Bruker Daltonics).

<p>ESBL: Extended-spectrum beta-lactamase, WT: wild type.</p

MLST allelic profile of <i>Klebsiella pneumoniae</i> clinical isolates distributed in the five clusters.

<p>ESBL: Extended-spectrum beta-lactamase, ESBL+Case: Extended-spectrum beta-lactamase associated to Cephalosporinase phenotype, Pase: Penicillinase phenotype, WT: Wild Type phenotype.</p

Data mining analysis of <i>Klebsiella pneumoniae</i> MSP dendrogram.

<p>WT: wild type phenotype, ESBL: Extended Spectrum Beta-lactamase<b>.</b> No represents the number of strains that correspond to the significant character according to data mining analysis.</p

Antibacterial resistance phenotypes of <i>Klebsiella pneumoniae</i> strains.

<p>ESBL: Extended-spectrum beta-lactamase, Case: Cephalosporinase, ESBL+Case: Extended-spectrum beta-lactamase associated to Cephalosporinase phenotype, Pase: Penicillinase, Pase IRT: inhibitor-resistant TEM penicillinase.</p

Geographic location of hospitals implicated in this study.

<p>1: Tlemcen, 2: Sidi Bel Abbes, 3: Oran, 4: Annaba, 5: Marseille, 6: Nice, 7: Angers.</p

Antibiotic susceptibility testing results of <i>Klebsiella pneumoniae</i> strains.

*<p>the <i>p</i> values compare the percentage of resistance and sensitivity between Algerian and French strains,</p><p><b>S</b>: Sensitive, <b>I</b> : Intermediate, <b>R</b> : Resistant, <b>AM</b>: Ampicillin, <b>AMX</b>: Amoxicillin, <b>AMC</b>: Amoxicillin/clavulanic acid, <b>TIC</b>: Ticarcillin, <b>CF</b>: Cefalotin, <b>FOX</b>: Cefoxitin, <b>CAZ</b>: Ceftazidime, <b>CTX</b>: Cefotaxime, <b>CRO</b>: Ceftriaxone, <b>IMP</b>: Imipenem, <b>GN</b>: Gentamicin, <b>AN</b>: Amikacin, <b>TM</b>: Tobramycin, <b>CIP</b>: Ciprofloxacin, <b>SXT</b>: Trimethoprim/Sulfamethoxazole, <b>CS</b>: Colistin.</p

Minimal spanning tree (MST) of <i>K. pneumoniae</i> isolates, showing relationships between STs, compared with clusters obtained from the dendrogram generated by BIOTYPER software.

<p>Minimal spanning tree (MST) of <i>K. pneumoniae</i> isolates, showing relationships between STs, compared with clusters obtained from the dendrogram generated by BIOTYPER software.</p

CNF1-triggered immunity requires inflammatory caspases.

<p>(A and B) IL-1β production and maturation/secretion after treatment with CNF1, LPS or CNF1 (1 μg/ml) + LPS (100 ng/ml) for 10 h. Actin and BSA were used as loading controls. (B) Graph showing the quantification of IL-1β secretion normalized to the control (n = 3). (C and D) Female C57BL/6 WT (C) or congenic C1^-/-C11^-/- mice (D) were intravenously infected with 10⁷ CFUs of <i>E</i>. <i>coli</i>^CNF1+ or with the isogenic CNF1-deleted mutant (<i>E</i>. <i>coli</i>^CNF1-) prior to the collection of peripheral blood at 3, 6, 24 and 48 h for bacteremia measurements. The data are expressed as the mean ± SEM (n = 7–8). (E and F) Analysis of the circulating levels of IL-1β (E) and KC (F) in the mouse sera by ELISA. Serum samples from mice infected with 10⁷<i>E</i>. <i>coli</i>^CNF1+ or with the isogenic mutant <i>E</i>. <i>coli</i>^CNF1- were collected at 3 h after intravenous infection and analyzed by ELISA (n = 3). P-values<0.05 (*); and P-values<0.01 (**) were considered statistically significant.</p

CNF1-triggered IL-1β maturation requires activated Rac, ASC and caspase-1.

<p>(A) Western blot analysis of the production and maturation/secretion of IL-1β by primary macrophages following treatment with CNF1, LPS or CNF1+LPS for 10 h. Actin and BSA were used as loading controls. (B) Quantification of caspase-1 activity in macrophages following treatment with CNF1+LPS for 6 h using YVAD-Fluorescent Labelled Inhibitor Caspase-1 Activity (FLICA). (C) Western blot analysis of macrophages IL-1β maturation/secretion upon transfection of HA-Rac2^Q61E and LPS treatment. (D) Co-immunoprecipitation of Myc-Rac2 and caspase-1 using an anti-Myc antibody following the treatment of HEK 293T cells with CNF1+LPS for 6 h.</p

CNF1 potentiates LPS-triggered immune responses.

<p>(A) Monocytes (5x10⁵ cells per condition) isolated from mouse blood were treated with PBS (control) or with 1 μg/ml CNF1 toxin for 10 h with or without 100 ng/ml LPS. Cell culture supernatants were analyzed using mouse ELISArray kits (n = 3). The data are shown as fold inductions compared with the control condition. (B, C and D) Monocytes (5x10⁵ cells per condition) purified from mouse blood were treated for 10 h. Cells were treated as indicated with 0.1, 1, or 10 μg/ml of CNF1 toxin or the CNF1 mutant C866S alone or in combination with ultrapure <i>E</i>. <i>coli</i> LPS at 1, 10 or 100 ng/ml (n = 3). (B) KC, (C) IL-6, and (D) IL-1β cytokine secretion was analyzed using ELISA (n = 4). (E) Female BALB/c mice were intravenously infected with 10⁷ CFUs of <i>E</i>. <i>coli</i> expressing CNF1 (<i>E</i>. <i>coli</i>^CNF1+) + PBS as a control or with an <i>E</i>. <i>coli</i>^CNF1+ + IL-1β antagonist (Kineret; 1.5 mg/kg) prior to the collection of peripheral blood at 3, 6, 24 and 48 h for the measurement of bacteremia (n = 10). P-values <0.05 (*); and P-values <0.01 (**) were considered statistically significant.</p