Role of the Corneal Epithelial Basement Membrane in Ocular Defense against Pseudomonas aeruginosa
- 1 August 2009
- journal article
- Published by American Society for Microbiology in Infection and Immunity
- Vol. 77 (8), 3264-3271
- https://doi.org/10.1128/iai.00111-09
Abstract
Pseudomonas aeruginosa can invade corneal epithelial cells and translocates multilayered corneal epithelia in vitro, but it does not penetrate the intact corneal epithelium in vivo. In healthy corneas, the epithelium is separated from the underlying stroma by a basement membrane containing extracellular matrix proteins and pores smaller than bacteria. Here we used in vivo and in vitro models to investigate the potential of the basement membrane to defend against P. aeruginosa . Transmission electron microscopy of infected mouse corneas in vivo showed penetration of the stroma by P. aeruginosa only where the basement membrane was visibly disrupted by scratch injury, suggesting that the intact basement membrane prevented penetration. This hypothesis was explored using an in vitro Matrigel Transwell model to mimic the corneal basement membrane. P. aeruginosa translocation of multilayered corneal epithelia grown on Matrigel was ∼100-fold lower than that of cells grown without Matrigel ( P < 0.005, t test). Matrigel did not increase transepithelial resistance. Matrigel-grown cells blocked translocation by a P. aeruginosa protease mutant. Without cells, Matrigel also reduced traversal of P. aeruginosa and the protease mutant. Fluorescence microscopy revealed a relative accumulation of bacteria at the superficial epithelium of cells grown on Matrigel at 3 h compared to cells grown on uncoated filters. By 5 h, bacteria accumulated beneath the cells, suggesting direct trapping by the Matrigel. These findings suggest that the basement membrane helps defend the cornea against infection via physical barrier effects and influences on the epithelium and that these roles could be compromised by P. aeruginosa proteases.Keywords
This publication has 44 references indexed in Scilit:
- Human Tear Fluid Protects against Pseudomonas aeruginosa Keratitis in a Murine Experimental ModelInfection and Immunity, 2007
- Corneal Virulence ofPseudomonas aeruginosaElastase B and Alkaline Protease Produced byPseudomonas putidaCurrent Eye Research, 2007
- The Pathogenesis of Contact Lens-Related KeratitisOptometry and Vision Science, 2006
- Factors associated with infection by Pseudomonas aeruginosa in adult cystic fibrosisEuropean Respiratory Journal, 2005
- A New Topical Model ofStaphylococcusCorneal Infection in the MousePublished by Association for Research in Vision and Ophthalmology (ARVO) ,2003
- Multidrug Efflux Systems Play an Important Role in the Invasiveness of Pseudomonas aeruginosa The Journal of Experimental Medicine, 2002
- Pseudomonas aeruginosaProteases and Corneal VirulenceDNA and Cell Biology, 2002
- Herpes simplex virus ophthalmic disease induced using two different methods of mice inoculationThe Brazilian Journal of Infectious Diseases, 2001
- Nanoscale topography of the basement membrane underlying the corneal epithelium of the rhesus macaqueCell and tissue research, 1999
- Evaluation of a Human Corneal Epithelial Cell Line as anin VitroModel for Assessing Ocular IrritationFundamental and Applied Toxicology, 1997