RRC ID 68722
Author Fukuda K, Ishida W, Tanaka H, Harada Y, Matsuda A, Ebihara N, Fukushima A.
Title Alarmins from corneal epithelial cells upregulate CCL11 and VCAM-1 in corneal fibroblasts.
Journal Invest Ophthalmol Vis Sci
Abstract PURPOSE:Severe ocular allergic diseases are characterized by pronounced conjunctival inflammation triggered by T helper 2 (Th2) cells and corneal epithelial damage induced by eosinophils. To examine the role of alarmins released by damaged corneal epithelial cells in tissue eosinophilia, we investigated the effects of a supernatant derived from necrotic human corneal epithelial (HCE) cells on expression of the chemokine CCL11 (eotaxin) and the adhesion molecule VCAM-1 in human corneal fibroblasts.
METHODS:An alarmin preparation was obtained as the material released from HCE cells after three cycles of freezing and thawing. CCL11 released into culture medium and cell surface expression of VCAM-1 were measured with enzyme-linked immunosorbent assays, and the amounts of CCL11 and VCAM-1 mRNAs were quantitated by reverse transcription and real-time polymerase chain reaction analysis. Signaling by the transcription factor NF-κB was evaluated by immunoblot and immunofluorescence analyses.
RESULTS:The combination of the necrotic HCE cell supernatant and either interleukin (IL)-4 or IL-13 induced synergistic increases in CCL11 release, VCAM-1 expression, and the abundance of CCL11 and VCAM-1 mRNAs in corneal fibroblasts. The necrotic HCE cell supernatant also induced NF-κB activation in corneal fibroblasts, whereas an inhibitor of NF-κB and IL-1 receptor antagonist each attenuated CCL11 release induced by the alarmin preparation and either IL-4 or IL-13.
CONCLUSIONS:Alarmins including IL-1 released from necrotic corneal epithelial cells cooperate with Th2 cytokines to induce CCL11 production and VCAM-1 expression in corneal fibroblasts, and may thereby play an important role in tissue eosinophilia associated with ocular allergic diseases.
Volume 54(8)
Pages 5817-23
Published 2013-8-27
DOI 10.1167/iovs.13-11969
PII iovs.13-11969
PMID 23920370
MeSH Chemokine CCL11 / metabolism* Cytokines / physiology* Enzyme-Linked Immunosorbent Assay Epithelium, Corneal / metabolism* Fibroblasts / metabolism* Humans Interleukin-13 / physiology Interleukin-4 / physiology Keratoconjunctivitis / metabolism* NF-kappa B / metabolism Necrosis / metabolism RNA, Messenger / metabolism Real-Time Polymerase Chain Reaction Signal Transduction / physiology Vascular Cell Adhesion Molecule-1 / metabolism*
IF 3.47
Human and Animal Cells HCE-T(RCB2280)