December 2002
Volume 43, Issue 13
Free
ARVO Annual Meeting Abstract  |   December 2002
Potential Role for Laminin-5 in Pseudomonas aeruginosa Binding to Extracellular Matrix Produced by Corneal Epithelialcells
Author Affiliations & Notes
  • MA Esco
    Anatomy and Cell Biology Wayne State University School of Medicine Detroit MI
  • S McClellan
    Anatomy and Cell Biology Wayne State University School of Medicine Detroit MI
  • LD Hazlett
    Anatomy and Cell Biology Wayne State University School of Medicine Detroit MI
  • M Kurpakus-Wheater
    Anatomy and Cell Biology Wayne State University School of Medicine Detroit MI
  • Footnotes
    Commercial Relationships   M.A. Esco, None; S. McClellan, None; L.D. Hazlett, None; M. Kurpakus-Wheater, None. Grant Identification: NIH EY02986, NIH P30EY04068
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 32. doi:
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      MA Esco, S McClellan, LD Hazlett, M Kurpakus-Wheater; Potential Role for Laminin-5 in Pseudomonas aeruginosa Binding to Extracellular Matrix Produced by Corneal Epithelialcells . Invest. Ophthalmol. Vis. Sci. 2002;43(13):32.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: To determine the role of laminin-5 in mediating the adhesion of Pseudomonas aeruginosa to extracellular matrix produced by cultured human corneal epithelial cells. To test the hypothesis that laminin-5 can facilitate bacterial adhesion to exposed extracellular matrix. Methods: Antibodies to the laminin beta3 chain (mAb17) and the laminin gamma2 chain (D4B5) were tested in adhesion assays for their ability to inhibit the binding of P. aeruginosa strain ATCC 19660 to extracellular matrix substrates (ECMS) produced by human corneal epithelial cells in culture. In one experiment mAb17 and D4B5 were mixed and incubated with the ECMS prior to application of ATCC 19660 suspended in D-PBS at pH 7.5. In a second experiment mAb17 and D4B5 were incubated with bacteria prior to application on the ECMS. As a control experiments were conducted concurrently using an irrelevant isotype-matched mAb. SEM and statistical analysis were performed to compare bacterial binding under experimental and control conditions. Results: Compared to irrelevant antibody control the presence of antibodies to the laminin-5 chains significantly increased the binding of ATCC 19660 to ECMS produced by human corneal epithelial cells. Significantly more binding was noted in the presence of the laminin-5 antibodies regardless of whether the antibodies were intially incubated on the ECMS prior to the application of bacteria or initially incubated with the bacteria prior to placement on the ECMS. Conclusion: We have previously shown that human corneal epithelial cells cultured in 2% oxygen (hypoxia) produce less laminin-5. We have also shown increased bacterial binding to the ECMS produced by hypoxic cells. This presents circumstantial evidence that the presence of laminin-5 in ECMS may actually be inhibitory to P. aeruginosa adhesion. This is supported by the antibody data presented in this abstract that demonstrates that inhibiting bacteria-laminin-5 interactions serves to enhance binding to ECMS.

Keywords: 531 Pseudomonas • 403 extracellular matrix • 372 cornea: epithelium 
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