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Synergistic Induction of Eotaxin Expression in Human Keratocytes by TNF- and IL-4 or IL-13

From the Department of Ophthalmology, Yamaguchi University School of Medicine, Japan.

PURPOSE. To investigate the effects of tumor necrosis factor (TNF)-, interleukin (IL)-4, and IL-13 on expression of the chemokine eotaxin by cultured human keratocytes.

METHODS. Cultured human keratocytes were incubated with various combinations and concentrations of TNF-, IL-4, and IL-13. The concentration of eotaxin in the culture supernatant was subsequently measured by enzyme-linked immunosorbent assay, and the amount of eotaxin mRNA in cell lysates was determined by reverse transcription–polymerase chain reaction analysis.

RESULTS. Keratocytes incubated in the absence of cytokines did not release detectable amounts of eotaxin into the culture medium. Whereas incubation of keratocytes with TNF-, IL-4, or IL-13 alone or with the combination of IL-4 and IL-13 had only a small effect on eotaxin release, exposure of the cells to TNF- in combination with either IL-4 or IL-13 resulted in a marked increase in eotaxin production that was both time and dose dependent. The abundance of eotaxin mRNA in keratocytes was also increased in a synergistic manner by incubation of cells with TNF- together with either IL-4 or IL-13.

CONCLUSIONS. Stimulation of human keratocytes with the combination of TNF- and either IL-4 or IL-13 resulted in synergistic increases in both the abundance of eotaxin mRNA and the release of eotaxin protein. This cytokine-induced increase in eotaxin production by keratocytes may contribute to eosinophil infiltration in inflammatory ocular diseases such as vernal keratoconjunctivitis.

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