Desiccating stress promotion of Th17 differentiation by ocular surface tissues through a dendritic cell-mediated pathway

Invest Ophthalmol Vis Sci. 2010 Jun;51(6):3083-91. doi: 10.1167/iovs.09-3838. Epub 2010 Feb 3.


Purpose: To explore the phenomenon that corneal and conjunctival tissues subjected to desiccating stress (DS) promote Th17 differentiation by stimulating the production of Th17-inducing cytokines through a dendritic cell (DC)-mediated pathway.

Methods: Experimental dry eye was created by subjecting C57BL/6 mice to desiccating environmental stress. Corneal and conjunctival explants from dry eye or control mice were cocultured with DCs for 24 hours before CD4(+) T cells were added for an additional 4 to 7 days. Expression of Th17-associated genes in the cornea, conjunctiva, DCs, and CD4(+) T cells was evaluated by real-time PCR. Cytokine concentrations in coculture supernatants were measured by immunobead assay. IL-17-producing T cells were identified by ELISPOT bioassay.

Results: Higher levels of IL-17A, TGF-beta1, TGF-beta2, IL-6, IL-23, and IL-1beta mRNA transcripts and TGF-beta1, IL-6, and IL-1beta protein were observed in corneal epithelium and conjunctiva from dry eye mice. DCs cocultured with epithelial explants from dry eye mice for 2 days produced higher levels of TGF-beta1, IL-6, IL-23, and IL-1beta mRNA transcripts and of TGF-beta1, IL-6, and IL-1beta protein. CD4(+) T cells cocultured with DCs and epithelial explants from dry eye mice expressed increased levels of IL-17A, IL-17F, IL-22, CCL-20, and retinoic acid receptor-related orphan receptor-gammat mRNA transcripts and increased IL-17A protein and number of IL-17-producing T cells (Th17 cells).

Conclusions: These findings demonstrate that DS creates an environment on the ocular surface that stimulates the production of Th17-inducing cytokines by corneal and conjunctival epithelia that promote Th17 differentiation through a dendritic cell-mediated pathway.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Marrow Cells
  • CD4-Positive T-Lymphocytes / immunology
  • Cell Differentiation*
  • Coculture Techniques
  • Conjunctiva / metabolism*
  • Cornea / metabolism*
  • Dendritic Cells / physiology*
  • Desiccation
  • Disease Models, Animal
  • Dry Eye Syndromes / chemically induced
  • Dry Eye Syndromes / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Epithelial Cells / metabolism
  • Female
  • Flow Cytometry
  • Gene Expression / physiology
  • Immunoenzyme Techniques
  • Interleukin-17 / genetics*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Reverse Transcriptase Polymerase Chain Reaction
  • Scopolamine / toxicity
  • T-Lymphocytes, Helper-Inducer / immunology*


  • Interleukin-17
  • Scopolamine