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1 From the Department of Ophthalmology, University of Washington School of Medicine, Seattle; the 2 Department of Ophthalmology, Korea University, Seoul; and the 3 Department of Ophthalmology, Research Institute of Medical Science, College of Medicine, Pusan National University, Korea.
PURPOSE. To determine the effect of interleukin (IL)-1
and tumor necrosis
factor (TNF)- on cytokine, chemokine, and receptor expression in
corneal stromal cells; the effect of corneal scrape injury on monocyte
chemotactic and activating factor (MCAF) expression and
monocyte-macrophage influx into the stroma; and the effect of MCAF and
granulocyte colony-stimulating factor (G-CSF) microinjection on
inflammatory cell infiltration into the stroma.
METHODS. Gene array technology was used to evaluate changes in cytokine,
chemokine, and receptor gene expression in stromal fibroblasts in
response to IL-1 and TNF. Expression of MCAF mRNA and protein was
monitored with an RNase protection assay and Western blot analysis,
respectively. Keratocyte MCAF protein expression in the rabbit cornea
was detected with immunocytochemistry. After epithelial scrape injury,
monocytes-macrophages were detected in rabbit corneas, by
immunocytochemistry for monocyte–macrophage antigen. Inflammatory cell
infiltration after MCAF and G-CSF microinjection into the stroma of
mouse corneas was monitored with hematoxylin and eosin staining.
RESULTS. IL-1 or TNF upregulated the expression of several proinflammatory
chemokines in stromal fibroblasts in culture. These included G-CSF,
MCAF, neutrophil-activating peptide (ENA-78), and monocyte-derived
neutrophil chemotactic factor (MDNCF). MCAF mRNA upregulation was
confirmed by RNase protection assay, and MCAF protein was detected by
Western blot analysis. MCAF protein was detected in keratocytes at 4
hours and 24 hours after epithelial injury, but not in keratocytes in
the unwounded cornea. Corneal epithelial injury triggered the influx of
monocytes-macrophages into the corneal stroma in the rabbit.
Microinjection of MCAF and G-CSF into mouse cornea resulted in the
influx of monocytes-macrophages and granulocytes, respectively, into
the stroma.
CONCLUSIONS. Proinflammatory chemokine induction in keratocytes is mediated by
IL-1 and TNF. The proinflammatory chemokines produced by the
keratocytes probably trigger the influx of inflammatory cells into the
stroma after epithelial injury associated with corneal surgery, contact
lenses, or trauma.
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