June 2020
Volume 61, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2020
Epithelium-derived interleukin-33 initiates mast cell-mediated corneal inflammation
Author Affiliations & Notes
  • Elsayed Elbasiony
    Schepens Eye Research Institute of Mass. Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Sharad Mittal
    Schepens Eye Research Institute of Mass. Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Wonkyung Cho
    Schepens Eye Research Institute of Mass. Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Sunil Chauhan
    Schepens Eye Research Institute of Mass. Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts, United States
  • Footnotes
    Commercial Relationships   Elsayed Elbasiony, None; Sharad Mittal, None; Wonkyung Cho, None; Sunil Chauhan, None
  • Footnotes
    Support  NIH/NEI-R01EY029727
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 463. doi:
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    • Get Citation

      Elsayed Elbasiony, Sharad Mittal, Wonkyung Cho, Sunil Chauhan; Epithelium-derived interleukin-33 initiates mast cell-mediated corneal inflammation. Invest. Ophthalmol. Vis. Sci. 2020;61(7):463.

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

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Abstract

Purpose : We have previously reported that injury-mediated activation of mast cells initiate corneal inflammation. In the present study, we sought to delineate the underlying mechanisms of mast cell activation following corneal injury that lead to neutrophil infiltration.

Methods : Potential mast cell activating factors (Interleukin (IL) 33, IL1β, IL6, SDF1α and Substance P) were used to stimulate mast cells for 3h. Mast cell activation markers β-tryptase and β-hexosaminidase and CXCL2 levels were evaluated in cultures using ELISA. Corneas were injured by removing the epithelium and anterior stroma with an Algerbrush II. IL33 levels were evaluated in tear wash of injured mice, as well as in epithelial and stromal lysates using ELISA. Infiltration of CD11b+Ly6G+ neutrophils (Median Fluorescent Intensity) was assessed in harvested corneas at 6h post-injury by flow cytometry. 2% Cromolyn or 1 mg/ml of IL33 neutralizing antibody were locally administered for in vivo mast cell inhibition.

Results : Significant mast cell activation and CXCL2 secretion were observed following IL33 stimulation, compared to other inflammatory factor. ELISA analysis showed higher levels of IL33 in (i) tear wash of injured mice compared to naive controls and (ii) epithelium compared to stroma (117.6 ± 5.8 vs. 28.5 ± 5.4 ng/ml; p<0.01). Corneal injury resulted in significantly increased levels of β-tryptase and β-hexosaminidase in tear wash (β-tryptase: 76.5 ± 19 vs. 14.3 ± 1.5 ng/ml; p<0.01 and β-hexosaminidase: 0.13 ± 0.001 vs. 0.05 ± 0.001 unit/ml; p<0.001) and higher frequencies of neutrophils at 6h post-injury, relative to naive corneas (4.6 ± 0.3% vs. 1.1 ± 0.2%; p<0.001). Topical blockade of IL33 significantly inhibited mast cell activation (β-tryptase: 19.2 ± 0.2 vs. 43.4 ± 8.7 ng/ml; p<0.01) and reduced early infiltration of neutrophils to the injured cornea, relative to control groups (1.6 ± 0.1% vs. 4.5 ± 0.6%; p<0.01).

Conclusions : Our data demonstrate that damaged corneal epithelial cells release pre-stored IL33, which activates mast cells to secrete CXCL2 initiating early neutrophil recruitment.

This is a 2020 ARVO Annual Meeting abstract.

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