Abstract
Purpose:
To test the hypothesis that the T helper (Th) 2 cytokine IL-13 promotes goblet cell proliferation and mucous production in primary mouse conjunctival cultures.
Methods:
Explants were obtained from the palpebral conjunctiva from young adult, (6-8 week) female C57BL/6 mice and plated one explant per well in 48 well plates. Explants received 200 µl per week of either Keratinocyte media with 3% fetal bovine serum defined (KSFM) or KSFM with 10ng/mL interleukin-13 (IL-13). Cultures were harvested on days 3, 7 or 14 for dot blot, RT-PCR or fixed in methanol and stained for Wheat Germ Agglutinin (WGA) glycoprotein, MUC5AC or cytokeratin 7 (K7).
Results:
Day 3 cultures treated with IL-13 illustrated smaller and more numerous cells. 100% of the cultures in all conditions were K7 positive and only K7 positive cells showed positive staining for WGA or MUC5AC. Increased production of MUC5AC was detected by dot blot at day 3 (P < 0.05) and at day 14 (P < 0.05) in IL-13 treated cultures compared to control. IL-13 treated cultures also demonstrated increased MUC5AC (P < 0.05) and K7 (P < 0.05) mRNA expression at day 14 compared to control.
Conclusions:
IL-13 cytokine stimulation increases epithelial proliferation of young (day 3) and MUC5AC production in young and mature (days 3 and 14) cultures. These experimental results suggest that the Th 2 cytokine IL-13 may play a vital role in conjunctival goblet cell growth and mucus production.
Keywords: 474 conjunctiva •
490 cytokines/chemokines •
541 glycoconjugates/glycoproteins