IL-13 Stimulates Proliferation and Expression of Mucin and Immunomodulatory Genes in Cultured Conjunctival Goblet Cells

Invest Ophthalmol Vis Sci. 2015 Jul;56(8):4186-97. doi: 10.1167/iovs.14-15496.

Abstract

Purpose: To investigate the effects of IL-13 on goblet cell proliferation, differentiation, and expression of mucin and immunomodulatory genes.

Methods: Explants were excised from the conjunctiva of young C57BL/6 mice. Cultures received 200 μL per week of either Keratinocyte media (KSFM) or KSFM supplemented with 10 ng/mL IL-13 and were incubated for 3 (D3), 7 (D7), or 14 (D14) days. Subsequently, cell proliferation was assessed or cultures were immunostained, collected for dot blot, or for reverse transcription (RT) and quantitative real-time PCR (qPCR) or for RT-PCR gene array.

Results: The cultured conjunctival epithelium expressed goblet cell associated keratin 7 and mucins MUC5AC and MUC2 and when stimulated with IL-13 showed increased proliferation at D3 and D7 (P < 0.05) compared with control. MUC5AC expression was increased in the IL-13-treated group at D3 and D14 (P < 0.05). IL-13-treated cultures showed increased chemokine ligand 26 (CCL26), chloride channel calcium activated channel 3 (CLCA3), fas ligand (FasL), and Relm-β at D7. All conjunctival cultures expressed MUC2, and its expression was decreased at D3 (P < 0.05) and increased at D14 (P < 0.05) with IL-13 treatment.

Conclusions: This study demonstrated that conjunctival goblet cells are IL-13 responsive cells that produce factors known to maintain epithelial barrier, stimulate mucin production, and modulate immune response in nonocular mucosa when treated with IL-13. The functional significance of IL-13-stimulated factors remains to be determined.

Publication types

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

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Differentiation
  • Cell Proliferation
  • Conjunctiva / metabolism*
  • Conjunctiva / pathology
  • Dry Eye Syndromes / genetics*
  • Dry Eye Syndromes / metabolism
  • Dry Eye Syndromes / pathology
  • Female
  • Gene Expression Regulation*
  • Goblet Cells / metabolism*
  • Goblet Cells / pathology
  • Interleukin-13 / biosynthesis
  • Interleukin-13 / genetics*
  • Mice
  • Mice, Inbred C57BL
  • Mucin 5AC / biosynthesis
  • Mucin 5AC / genetics*
  • RNA, Messenger / genetics*
  • Real-Time Polymerase Chain Reaction

Substances

  • Interleukin-13
  • Muc5ac protein, mouse
  • Mucin 5AC
  • RNA, Messenger