June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Alarmin from corneal epithelial cells synergizes with Th2 cytokines on the expression of CCL11 and VCAM-1 by corneal fibroblasts
Author Affiliations & Notes
  • Ken Fukuda
    Ophthalmology, Kochi Medical School, Nankoku, Japan
  • Hiroshi Tanaka
    Ophthalmology, Machida Hospital, Kochi, Japan
  • Waka Ishida
    Ophthalmology, Kochi Medical School, Nankoku, Japan
  • Yosuke Harada
    Ophthalmology, Kochi Medical School, Nankoku, Japan
  • Tamaki Sumi
    Ophthalmology, Kochi Medical School, Nankoku, Japan
  • Akira Matsuda
    Ophthalmology, Juntendo University School of Medicine, Tokyo, Japan
  • Nobuyuki Ebihara
    Ophthalmology, Juntendo University School of Medicine, Tokyo, Japan
  • Atsuki Fukushima
    Ophthalmology, Kochi Medical School, Nankoku, Japan
  • Footnotes
    Commercial Relationships Ken Fukuda, None; Hiroshi Tanaka, None; Waka Ishida, None; Yosuke Harada, None; Tamaki Sumi, None; Akira Matsuda, None; Nobuyuki Ebihara, None; Atsuki Fukushima, Otsuka Pharmatheutical Co., Ltd (F)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 5244. doi:https://doi.org/
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      Ken Fukuda, Hiroshi Tanaka, Waka Ishida, Yosuke Harada, Tamaki Sumi, Akira Matsuda, Nobuyuki Ebihara, Atsuki Fukushima; Alarmin from corneal epithelial cells synergizes with Th2 cytokines on the expression of CCL11 and VCAM-1 by corneal fibroblasts. Invest. Ophthalmol. Vis. Sci. 2013;54(15):5244. doi: https://doi.org/.

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

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Abstract

Purpose: Clinical features of severe ocular allergic disease, vernal keratoconjunctivitis, are the severe conjunctival inflammation and the corneal epithelial damage due to eosinophils which results in visual disturbance. Corneal stromal fibroblasts are the main source of CCL11/eotaxin-1. To know the role of alarmins from damaged corneal epithelial cells in tissue eosinophilia, we have now examined the effects of the supernatant derived from necrotic corneal epithelial cell and Th2 cytokines on corneal fibroblasts.

Methods: Necrosis of cultured human corneal epithelial cells was induced by repeated freezing and thawing. The amount of chemokines released into culture medium by human corneal fibroblasts was determined by enzyme-linked immunosorbent assay, surface expression of adhesion molecules on the cultured cells was measured with a whole-cell enzyme-linked immunosorbent assay, and the intracellular abundance of these molecules mRNA was quantitated by reverse transcription and real-time polymerase chain reaction analysis. Signaling by the transcription factors NF-κB was evaluated by immunoblot and immunofluorescence analyses.

Results: The supernatant from necrotic corneal epithelial cells induced a low level of CCL11 release by corneal fibroblasts, as did Th2 cytokines, IL-4 and IL-13. However, the combination of the supernatant from necrotic corneal epithelial cells and IL-4 or IL-13 induced a marked synergistic increase in CCL11 release. The abundance of CCL11 mRNA in the fibroblasts was also increased in a synergistic manner by costimulation with the supernatant from necrotic corneal epithelial cells and IL-4 or IL-13. In addition, the combination of the supernatant from necrotic corneal epithelial cells and IL-4 or IL-13 synergistically increased the expression of VCAM-1 in corneal fibroblasts. The supernatant from necrotic corneal epithelial cells activated NF-κB in corneal fibroblasts, and inhibitors of NF-κB and IL-1 receptor antagonist attenuated CCL11 release and VCAM-1 expression induced by the supernatant from necrotic corneal epithelial cells in combination with IL-4 or IL-13.

Conclusions: The interaction between IL-1 from necrotic corneal epithelial cells and Th2 cytokines may play an important role in tissue eosinophilia associated with ocular allergic diseases.

Keywords: 480 cornea: basic science • 484 cornea: stroma and keratocytes • 490 cytokines/chemokines  
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