May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
A Bacillus cereus Secreted Factor InducesPermeability of an in vitro Blood Retinal Barrier
Author Affiliations & Notes
  • A. L. Moyer
    Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, Oklahoma
    Microbiology,
  • J. Thurman
    Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, Oklahoma
    Department of Ophthalmology,
  • M. C. Callegan
    Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, Oklahoma
    Department of Ophthalmology,
    Department of Ophthalmology, Dean A. McGee Eye Institue, Molecular Pathogenesis of Eye Infections Research Center, Oklahoma City, Oklahoma
  • Footnotes
    Commercial Relationships  A.L. Moyer, None; J. Thurman, None; M.C. Callegan, Advanced Medical Optics, Allergan, Alcon, C.
  • Footnotes
    Support  EY12985 NIH/NEI (Callegan), EY12190 NIH/NCCR CORE Grant (Anderson), Research to Prevent Blindness Lew R. Wasserman Award (Callegan), Research to Prevent Blindness Unrestricted Research Grant (DMEI)
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 2481. doi:https://doi.org/
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    • Get Citation

      A. L. Moyer, J. Thurman, M. C. Callegan; A Bacillus cereus Secreted Factor InducesPermeability of an in vitro Blood Retinal Barrier. Invest. Ophthalmol. Vis. Sci. 2008;49(13):2481. doi: https://doi.org/.

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

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Abstract

Purpose: : Bacillus endophthalmitis causes permeability of the blood-retinal barrier (BRB). We sought to determine whether permeability was caused by a secreted bacterial factor and characterize the factor.

Methods: : An in vitro BRB consisting of polarized RPE monolayers was cultivated on collagen-, fibronectin-, and protein-coated glass coverslips and incubated with 102 CFU/ml of wild type or plcR-deficient B. cereus, sterile B. cereus supernatants, heat inactivated supernatants, purified cell walls, or 102 CFU/ml B. subtilis. Permeability was measured by dextran conjugate transmigration assay, transepithelial resistance, and ZO-1 immunocytochemistry. Supernatants prepared in BHI and RPE media were analyzed by silver stain, zymogram, and hide azure blue assay to analyze for the presence of proteases.

Results: : Incubation of polarized RPE with supernatants of wild typeor plcR-deficient B. cereus resulted in similar degrees of permeability by 8 h, with wild type Bacillus supernatant causing more rapid permeability than the plcR-deficient supernatant. PlcR-deficient supernatant caused alterations in ZO-1 as early as 6 h, while wild type supernatant caused ZO-1 alterations after 6 h. Neither heat inactivated supernatants, cell wall preparations, nor the growth of B. subtilus caused RPE monolayer permeability. The plcR-deficient strain retained significant protease activity in both BHI and RPE media, compared to wild type controls.

Conclusions: : B. cereus infection of an in vitro BRB resulted in permeability that was independent of plcR-regulation and caused by a secreted factor. Future studies will focus on confirming the identity of the permeability factor and identifying the mechanism of BRB breakdown during B. cereus endophthalmitis.

Keywords: endophthalmitis • bacterial disease • retinal pigment epithelium 
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