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Differential and Cooperative Effects of TNF, IL-1ß, and IFN on Human Conjunctival Epithelial Cell Receptor Expression and Chemokine Release

¹From the Departments of Medicine and ³Ophthalmology and Visual Sciences, School of Medicine, University of Wisconsin-Madison, Madison, Wisconsin.

PURPOSE. To gain better understanding of conjunctival epithelial cell responses to proinflammatory cytokines, the individual and combined effects of TNF, IL-1ß, and IFN on chemokine release (IL-8, regulated on activation normal T-cell expressed and secreted [RANTES]) and surface receptor expression (intercellular adhesion molecule [ICAM]-1, and HLA-DR, -DP, and -DQ) were examined.

METHODS. Conjunctival epithelial cells were isolated from cadaveric conjunctival tissues and cultured in 24-well plates until almost confluent. Recombinant cytokines (0.005–50 ng/mL) were added, alone or in various combinations, 24 hours before harvesting of supernates for ELISAs and cells for flow cytometry.

RESULTS. TNF, IL-1ß, and IFN had distinctive individual and combined effects on the parameters tested. Although TNF and IL-1ß had similar and synergistic effects on increasing expression of ICAM-1, IL-1ß was a more potent upregulator of the release of IL-8 than was TNF. Upregulation of IL-8 was additive when IL-1ß was combined with TNF. Neither TNF nor IL-1ß increased expression of HLA. In contrast, IFN was a potent upregulator of both surface receptors (ICAM-1 and HLA) but IFN alone had no effect on mediator release (IL-8 and RANTES). Release of RANTES required two cytokine signals, with IFN and TNF being the most potent combination.

CONCLUSIONS. Knowledge of the differential and combined effects of proinflammatory cytokines on conjunctival epithelial cells allows better understanding of ocular inflammation.

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