June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
The role of type 4 pilus, flagella and type III secretion system in the interactions between Pseudomonas aeruginosa bacteria and corneal fibroblasts in human microbial keratitis
Author Affiliations & Notes
  • Ahmad Elsahn
    Clinical Experimental Sciences, University of Southampton, Southampton, United Kingdom
    University Hospitals Southampton, Southampton, United Kingdom
  • Myron Christodoulides
    Clinical Experimental Sciences, University of Southampton, Southampton, United Kingdom
  • Parwez Hossain
    Clinical Experimental Sciences, University of Southampton, Southampton, United Kingdom
    University Hospitals Southampton, Southampton, United Kingdom
  • Footnotes
    Commercial Relationships   Ahmad Elsahn, None; Myron Christodoulides, None; Parwez Hossain, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 5765. doi:
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    • Get Citation

      Ahmad Elsahn, Myron Christodoulides, Parwez Hossain; The role of type 4 pilus, flagella and type III secretion system in the interactions between Pseudomonas aeruginosa bacteria and corneal fibroblasts in human microbial keratitis
      . Invest. Ophthalmol. Vis. Sci. 2017;58(8):5765.

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

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Abstract

Purpose : To examine the specific individual roles of bacterial virulence factors in the interactions between Pseudomonas aeruginosa (PA) bacteria and corneal fibroblasts in human microbial keratitis.

Methods : Human corneal fibroblast (CF) were extracted from clinical samples from several patients and cultured to confluence in vitro. CFs were challenged with live wild type PAO1 bacteria and mutant strains deficient in type 4 pilus (TFP), flagella or both, and total bacterial association was quantified at 3h using the saponin-lysis and viable counting method and confirmed using laser scanning confocal microscopy (LSCM); and live wild type PA14 bacteria and mutant strains deficient in type III secretion system (TTSS) or flagella, and lactose dehydrogenase (LDH) release from CF was quantified and used as a measure of cytotoxicity.

Results : Bacterial adherence to CF was significantly less (P=0.000, ANOVA) in mutant PAO1 strains deficient in TFP and flagella compared to wild type strain as demonstrated by viable counting assays and LSCM. Bacteria-induced cytotoxicity was significantly less (P=0.000, ANOVA) in mutant strains deficient in TTSS or flagella compared to wild type strain as demonstrated by reduced LDH release.

Conclusions : We have established in vitro cell culture models of bacterial keratitis and demonstrated that PA utilises TFP and flagella to adhere to, and TTSS to induce cytotoxicity in, human CF.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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