Adhesion patterns of commensal and pathogenic Escherichia coli from humans and wild animals on human and porcine epithelial cell lines - PubMed (original) (raw)

doi: 10.1186/1757-4749-5-31.

Alexander Böhm, Jörg Nitschke, Jörg Weinreich, Julia Groß, Stefan Rödiger, Thomas Wex, Hermann Ansorge, Olaf Zinke, Christian Schröder, Dirk Roggenbuck, Peter Schierack

Affiliations

Adhesion patterns of commensal and pathogenic Escherichia coli from humans and wild animals on human and porcine epithelial cell lines

Ulrike Frömmel et al. Gut Pathog. 2013.

Abstract

Background: Different strategies of colonization or infection by E. coli result in formation of certain adhesion patterns which help also in classifying intestinal E. coli into pathotypes. Little is known about adhesion patterns and host- and tissue adaption of commensal E. coli and about E. coli originating in clinically healthy hosts carrying pathotype-specific virulence-associated genes.

Findings: Adhesion pattern of E. coli (n = 282) from humans and from 18 animal species were verified on intestinal human Caco-2 and porcine IPEC-J2 cells and, furthermore, for comparison on human urinary bladder 5637, porcine kidney PK-15 epithelial and HEp-2 cells. The analysis was carried out on 150,000 images of adhesion assays.Adhesion patterns were very diverse; 88 isolates were completely non-adherent, whereas 194 adhered to at least one cell line with the dominant adhesion patterns "diffusely distributed" and "microcolony formation". Adhesion patterns "chains" and "clumps" were also visible. Chain formation was mediated by the presence of epithelial cells. Clump formation was very specific on only the 5637 cell line. All enteropathogenic (eae+) E. coli (EPEC; n = 14) were able to form microcolonies which was cell line specific for each isolate. Most EPEC formed microcolonies on intestinal IPEC-J2 and Caco-2 but several also on urinary tract cells. Shigatoxin-producing (stx+) E. coli (n = 10) showed no specific adhesion patterns.

Conclusions: E. coli isolates were highly diverse. Commensal and pathogenic isolates can adhere in various forms, including diffuse distribution, microcolonies, chains and clumps. Microcolony formation seems to be a global adhesion strategy also for commensal E. coli.

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Figures

Figure 1

Figure 1

E. coli adhesion patterns of commensal E. coli and tEPEC. Fluorescent images of epithelial cells and adherent bacteria which were stained with propidium iodide are depicted. A)E. coli isolate from a domestic pig adhering diffusely distributed to IPEC-J2 cells. B)E. coli isolate from a European hare forming microcolonies on Caco-2 cells. C)E. coli isolate from a human forming clumps on 5637 cells. D)E. coli isolate from a common blackbird forming microcolonies on 5637 cells. E)E. coli isolate from a wild boar forming chains on PK-15 cells. F) tEPEC isolate from a European hedgehog forming microcolonies on 5637 cells. Scale: 10 μm.

Figure 2

Figure 2

Chain formation of one E. coli isolate from a common blackbird. Fluorescence images were taken after incubation in LB medium (overnight), cell culture media (four hours) and after incubation on epithelial cells (four hours). Chain formation is induced by cell culture media, and stimulated by epithelial cells. Isolate in A) LB medium: no chains, B) Caco-2 cell culture medium, C) 5637 cell culture medium, D) IPEC-J2 and PK-15 cell culture medium. Isolate after four-hour adhesion assay with cell culture media on E) Caco-2, F) 5637, G) PK-15 cells. H) Isolate after four-hour adhesion assay with LB media on IPEC-J2 cells. A- D) Phase contrast microscopy. E- H) Fluorescence microscopy. Scale: 10 μm.

Figure 3

Figure 3

Adhesion pattern formation of 14 eae A + E. coli isolates on four epithelial cells and on HEp- 2 cells after six- hour incubation. Microcolonies were formed on IPEC-J2 (10 aEPEC, 2 tEPEC), Caco-2 (10 aEPEC, no tEPEC), HEp-2 (7 aEPEC, 1 tEPEC), PK-15 (2 aEPEC, 1 tEPEC) and 5637 cells (2 aEPEC, no tEPEC).

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