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Rebecca A. Fillmore, Shannon E. Nelson, Robert N. Lausch, John E. Oakes; Differential Regulation of ENA-78 and GCP-2 Gene Expression in Human Corneal Keratocytes and Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2003;44(8):3432-3437. doi: 10.1167/iovs.03-0095.
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purpose. To determine whether interleukin (IL)-1α- and tumor necrosis factor (TNF)-α–stimulated human corneal epithelial cells (HCECs) and human corneal keratocytes (HCKs) produce the α-chemokines epithelial cell-derived neutrophil attractant (ENA)-78 and granulocyte chemotactic protein (GCP)-2.
methods. Cultures of HCECs and HCKs were stimulated with either human recombinant IL-1α or TNF-α. At selected times after stimulation, culture supernatants were harvested and assayed for ENA-78 and GCP-2 by enzyme-linked immunosorbent assay. RNA was extracted from cell cultures to measure steady state levels of intracellular ENA-78 and GCP-2 pre-mRNA and mRNA by the reverse transcription–polymerase chain reaction.
results. Exposure of HCECs to either IL-1α or TNF-α stimulated a more than 4.5-fold increase in ENA-78 RNA and protein synthesis without stimulating a significant increase in either GCP-2 RNA synthesis or protein production. Exposure of HCK to IL-1α stimulated a 10-fold increase in ENA-78 and GCP-2 RNA synthesis and a more than 300-fold increase in ENA-78 and GCP-2 protein production. In contrast, exposure of keratocytes to TNF-α significantly enhanced ENA-78 RNA synthesis, resulting in a more than 68-fold increase in ENA-78 protein synthesis without significantly enhancing either GCP-2 gene expression or protein secretion.
conclusions. ENA-78 gene expression is significantly enhanced in both HCECs and HCKs in response to either IL-1α or TNF-α stimulation. In contrast, GCP-2 synthesis is only inducible in IL-1α–stimulated HCKs. The results suggest that GCP-2 gene expression is more tightly regulated in diseased or injured corneal tissue than is ENA-78 gene expression.
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