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1From the Department of Ophthalmology and Visual Science, University of Chicago, Chicago, Illinois; the 2Kresge Eye Institute, Wayne State University, Detroit, Michigan; and the 3Department of Environmental Health, School of Public Health, Boston University, Boston, Massachusetts.
PURPOSE. To examine the regulatory effects of interferon (IFN)-
, IFN-
, transforming growth factor (TGF)-ß, and tumor necrosis factor (TNF)-
on human fetal retinal pigment epithelial (HFRPE) cellinduced apoptosis of human Jurkat T (Jkt) cells.
METHODS. Pure cultures of HFRPE cells were isolated. The cells were precultured with medium alone or with addition of IFN-
, IFN-
, TNF-
, or TGF-ß for 72 hours. Thereafter, HFRPE cells were extensively washed before they were cocultured jointly with Jkt cells (standard) or cultured alone for another 48 hours to accumulate conditioned medium that is collected and added as cell-free conditioned medium to Jkt cell cultures (supernatant). Jkt cells were cocultured under the two culture conditions for 48, 72, and 96 hours. The rate of apoptosis in Jkt T cells was determined with annexin V staining and flow cytometry.
RESULTS. Both IFN-
and -
upregulated HFRPE-induced apoptosis in Jkt T cells. However, the apoptosis induced by IFN-
activated HFRPE cells was significant only in the absence of cellcell contact (supernatant). The supernatant induced a higher rate of apoptosis in Jkt T cells when compared to the direct coculture of the cells. TGF-ß and TNF-
did not upregulate HFRPE-induced apoptosis in Jkt T cells.
CONCLUSIONS. These results indicate that type I and type II IFNs can upregulated HFRPE-induced apoptosis in Jkt T cells, IFN-
being the more effective cytokine. Neither, TGF-ß nor TNF-
upregulated the HFRPE-induced apoptosis in Jkt T cells. Although HFRPE-induced apoptosis was mediated in a cellcell-contactindependent pathway, HFRPE cells may also express membrane-bound antiapoptotic molecules. These findings may help us to understand better the modulatory effects of pro- and anti-inflammatory cytokines on immune suppressive characteristics of RPE cells.
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