Exposure of a corneal epithelial cell line (hTCEpi) to Demodex-associated Bacillus proteins results in an inflammatory response

Invest Ophthalmol Vis Sci. 2014 Oct 2;55(10):7019-28. doi: 10.1167/iovs.14-15018.

Abstract

Purpose: A role for a bacterium, Bacillus oleronius, originally isolated from a Demodex mite, in the induction of ocular rosacea has been proposed. The aim of this work was to characterize the response of a corneal epithelial cell line to Bacillus proteins, as this might give an insight into how such proteins contribute to the symptoms of ocular rosacea in vivo.

Methods: The effect of exposing Bacillus protein preparation on human telomerase-immortalized corneal epithelial cells (hTCEpi) was measured by monitoring changes in cell proliferation and the expression of a number of genes associated with inflammation. The production of inflammatory cytokines was measured and the expression and activity of MMP-9 was quantified.

Results: Exposure of hTCEpi cells to 2 or 6 μg/mL Bacillus protein resulted in a dose-dependent reduction in cell proliferation. Exposure of cells to 6 μg/mL Bacillus protein did not induce apoptosis, but there was an increase in the expression of genes coding for IL-6 (13.8-fold), IL-1β (4.0-fold), IL-8 (11.1-fold), and TNF-α (4.1-fold). Increased expression of genes coding for the defensins, CCL20 (4.5-fold) and S100A7 (6.8-fold) also was observed. Elevated production of IL-6 and IL-8 was evident from cells exposed to 2 and 6 μg/mL Bacillus protein. The hTCEpi cells demonstrated increased MMP-9 expression (3.2-fold, P = 0.003) and activity (2.2-fold, P = 0.0186) after 48 hours of exposure to 6 μg/mL Bacillus protein preparation.

Conclusions: The results suggest that interaction of Demodex-associated Bacillus proteins with the corneal surface could lead to tissue degradation and inflammation, possibly leading to corneal scarring.

Keywords: Bacillus; Demodex; corneal scarring; corneal surface; ocular rosacea; sterile ulcers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus / isolation & purification
  • Bacillus / metabolism*
  • Bacterial Proteins / metabolism*
  • Blotting, Western
  • Cell Cycle
  • Cell Line
  • Cell Proliferation
  • Corneal Ulcer / metabolism*
  • Corneal Ulcer / microbiology
  • Corneal Ulcer / pathology
  • Enzyme-Linked Immunosorbent Assay
  • Epithelium, Corneal / metabolism*
  • Epithelium, Corneal / microbiology
  • Epithelium, Corneal / pathology
  • Eye Infections, Bacterial / metabolism*
  • Eye Infections, Bacterial / microbiology
  • Eye Infections, Bacterial / pathology
  • Humans

Substances

  • Bacterial Proteins