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. 2000 Dec 29;2(1):10–19. doi: 10.1186/rr33

Figure 1.

Figure 1

Ex vivo infection of human tracheal explants and infection of human tracheal epithelial cells. (a) Pieces (1 cm3) of freshly isolated normal adult human trachea were infected with the Escherichia coli isolates NU14 (fimH+) and NU14-1 (fimH-) for 90 min and incubated for 24 h in fresh media containing 50 μg/ml gentamicin as previously described [40]. NU14 is an E. coli strain isolated from a patient with cystitis and expresses FimH-containing type 1 pili. NU14-1 is a fimH- mutant in which a chloramphenicol cassette was recombined into the fimH gene in the chromosome of NU14 [25]. FimH is a mannose-binding adhesin that mediates the interaction of type 1-piliated bacteria with mannose-containing glycoproteins on eukaryotic cell surfaces. Released and activated matrilysin was detected by western analysis. (b) Human tracheal primary epithelial cells were infected with the type 1 piliated recombinant strains ORN103/pSH2 (fimH+) and ORN103/pUT2002 (fimH-) for 90 min, and allowed to condition fresh media for 48 h. Matrilysin secretion was assessed by immunoblotting. (Reproduced from [40]; copyright permission of The Rockefeller University Press.)