May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Human Limbal Basal Epithelium-Derived GDNF Suppresses the Stress and Inflammation-Induced Th17 Cytokines Through NF-kB Signaling Pathway
Author Affiliations & Notes
  • F. Bian
    Ocular Surface Center, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas
    Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
  • H. Qi
    Ocular Surface Center, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas
    Peking University Eye Center, Peking University Third Hospital, Beijing, China
  • S. C. Pflugfelder
    Ocular Surface Center, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas
  • D.-Q. Li
    Ocular Surface Center, Department of Ophthalmology, Baylor College of Medicine, Houston, Texas
  • Footnotes
    Commercial Relationships  F. Bian, None; H. Qi, None; S.C. Pflugfelder, None; D. Li, None.
  • Footnotes
    Support  DOD CDMRP Grant FY06 PR064719 (DQL), NIH Grants, EY014553 (DQL) and EY11915 (SCP), Lions Eye Bank Foundation grant (DQL), Research to Prevent Blindness, Oshman Foundation, William Stamps Farish Fund.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 4520. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      F. Bian, H. Qi, S. C. Pflugfelder, D.-Q. Li; Human Limbal Basal Epithelium-Derived GDNF Suppresses the Stress and Inflammation-Induced Th17 Cytokines Through NF-kB Signaling Pathway. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4520. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : The immune defensive privilege of corneal epithelial stem cells is not clear. This study was to explore the immune defensive role of glial cell-derived neurotrophic factor (GDNF) in protecting human limbal stem cells against stress and inflammation-stimulated Th17 pathway.

Methods: : Fresh human corneoscleral cryosections were used for immunostaining. Primary human corneal epithelial cultures were treated with hyperosmolar media, TNF-α, IL-1β or IL-17A, with or without GDNF or NF-kB inhibitors for 2-24 hours. The mRNA expression was determined by Affymetrix microarray and real time PCR with TaqMan primers and probes. NF-kB activation was detected by immunostaining, Western blot and cell-bases activation ELISA.

Results: : GDNF and its receptor GFRα-1 were found to be exclusively localized in the basal layer of limbal epithelium where stem cells reside. Affymetrix microarray revealed the Th17 family cytokines IL-17C, IL-17D (but not IL-17A, B, and F) and their receptors, IL-17RA, IL-17RB, IL-17RC and IL-17RD, were expressed by normal corneal epithelia. IL-17RA was immuno-localized in most corneal and limbal epithelial layers, but was negative in the limbal basal cells. The hyperosmotic (400-500 mOsM) stress and inflammatory cytokines TNF-α and IL-1β significantly increased mRNA levels of IL-6 and TGF-β2 (inducers for TH17 initiation) as well as IL-23 and IL-15 (inducers for Th17 survival and expansion), and further induced IL-17F expression in human corneal epithelial cells. Hyperosmotic stress or TNF-α activated NF-kB p65 nuclear translocation and Ser536 phosphorylation. GDNF, NF-kB inhibitor (quinazoline) or IKK inhibitor II (Wedelolactone) blocked the NF-kB activation, and suppressed stimulated production of these Th17 inducers and IL-17F by hyperosmolarity and TNF-α. Exogenous IL-17A promoted the expression of TNF-α, IL-1β, IL-8, MMP-9 and MMP-3. IL-17A also stimulated Th17 inducers IL-6, IL-23 and IL-15. Interestingly, GDNF partially suppressed these inflammatory responses to Th17 cytokine.

Conclusions: : These findings demonstrate that limbal basal epithelial-expressed GDNF suppresses stress and inflammation-induced Th17 cytokines by inhibiting the NF-kB signaling pathway. GDNF also partially suppressed the IL-17 induced inflammatory response, which protects corneal epithelial stem cells against inflammatory insults.

Keywords: cornea: epithelium • immune tolerance/privilege • signal transduction 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×