Effects of reduced mucus oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients
J. Clin. Invest. Dieter Worlitzsch, et al. 109:317 doi:10.1172/JCI13870 [
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Figure 1P. aeruginosa is localized in intraluminal material of freshly excised CF airways and binds to mucus. (
a) Thin section of an obstructed CF bronchus, stained with hematoxalin/eosin. Note the absence of
P. aeruginosa on epithelial surface (black arrow) and presence of
P. aeruginosa macrocolonies within intraluminal material (white arrows). Blue gap is an artifact due to fixation. (
b)
P. aeruginosa within macrocolonies in a lung section, stained with rabbit Ab’s against
P. aeruginosa. Bars:
a, 100 μm;
b, 10 μm. (
c) Percentage of bacteria detected at a distance of 2-5 μm or 5-17 μm from the epithelial surface of lungs from nine CF patients. Shrinkage artifacts were subtracted from calculated distances. (
d) Scanning electron micrograph of mucus-coated spheroid derived from CF respiratory epithelium.
P. aeruginosa (white arrow) were enmeshed in mucus (black arrows) following a 2-hour incubation. (
e) Immunofluorescent staining of mucins (anti-mucin Ab) bound to
P. aeruginosa strain PAO1 in vitro. (
f) Spheroid with adherent mucus removed by prewash, then incubated with
P. aeruginosa for 2 hours. Note the absence of bacteria on ciliated epithelial cell surfaces. Bars:
d, 0.6 μm;
e, 4 μm;
f, 2.5 μm. Quantitative comparisons of PAO1 binding revealed higher binding to mucus-coated NESfrom normal subjects (21.3 ± 10.6 bacteria/NES) versus non-mucus-coated (washed) NES (7.1 ± 0.1 bacteria/NES) (
n = 6; 3 normal subjects;
P < 0.05). Importantly, these values were not different for CF NESs (26.4 ± 4.1 bacteria/NES for mucus-coated NESs; 7.7 ± 3.9 bacteria/NES for washed NESs).
