coli strains from clusters 1 to 4 Antimicrobial sensitivity patt

coli strains from clusters 1 to 4. Antimicrobial sensitivity patterns did not differ significantly between phylogenetic groups (Fig. S4). However, E. coli from MLST clusters 2, 3 and 4 were more likely to have resistance to at least one antimicrobial than cluster 1 bacteria (cluster 1, 0/5; cluster selleckchem Ponatinib 2, 2/7; cluster 3, 2/7; cluster 4, 7/7; P<0.05). Oxytetracycline is widely used to treat PID and resistance to oxytetracycline also differed amongst the E. coli (cluster 1, 0/5; cluster 2, 0/7; cluster 3, 2/7; cluster 4, 7/7; P<0.05). E. coli or LPS Infused into the Uterus of Mice Causes PID or Endometritis Live bacteria and purified LPS from MLST cluster 1 and 4 E. coli were infused into the uterine lumen of C57BL6 mice.

Within 24 h of infusion 5/5 mice administered the cluster 4 O73:H16 bacteria showed signs of toxaemia (cold extremities, reduced appetite, inactivity), one animal had a uterus distended with purulent material and one animal died. No clinical signs were observed in animals administered cluster 1 O(84,172):H+ bacteria (n=5), LPS from either cluster (n=5 per cluster) or vehicle (n=5). The endometrium from the mice infused with live bacteria or LPS was inflamed with evidence of neutrophil accumulation (Fig. 6A�CC). Immunhistochemistry confirmed the location of neutrophils (Fig. 6D�CF) and macrophages (Fig. 6G�CI) with the immune cells localised to the epithelium and lumen in mice infused with LPS but more widespread in the endometrium of mice infused with live bacteria. E.

coli were detectable by fluorescence in situ hybridization (FISH) colonising the endometrium only in animals infused with bacteria, and could be seen invading throughout the endometrium and myometrium (Fig. 6J�CL). Figure 6 Uterine-derived E. coli cause PID in mice. LPS from Uterine E. coli Provokes Endometrial Cell Inflammation via TLR4 The key receptor for LPS on professional immune cells is TLR4, leading to activation of immune and inflammatory pathways [17]. To test the role of TLR4 in the endometrium, epithelial and stromal cells from C57Bl6 wild type or TLR4?/? mice were challenged with LPS purified from MLST cluster 4 E. coli, using commercially purified LPS as a positive control. As for the bovine cells, the response to LPS was tested by measuring the accumulation in the medium of prostaglandin PGE and a chemokine similar to IL-8, chemokine (C-X-C motif) ligand 1 (CXCL1; also known as keratinocyte-derived chemokine, KC).

Epithelial and stromal Cilengitide cells from wild type but not TLR4?/? mice secreted PGE and CXCL1 in response to LPS (Fig. 7). Figure 7 Endometrial cells respond to LPS and the response is TLR4 dependent. Discussion Gram-negative infections of the upper female genital tract are an important cause of PID and infertility in humans and animals [2], [9]. The present study focussed on E.

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