Changes in the mRNA expression of cytokines and chemokines by stimulated RPE cells in vitro.
OBJECTIVE
The transplantation of retinal pigment epithelial (RPE) cells is a possible therapy for degenerative diseases of the retina. However, the immune response and the subsequent rejection of the allografts are major problems in this field. We investigated the effect of pro-inflammatory factors on the cytokine and chemokine mRNA expression of human RPE cells during long-term observations in vitro.


METHODS
Human RPE cells were cultured in the presence of tumor necrosis factor-alpha (TNF-alpha, 10 ng/ml), interferon-gamma (IFN-gamma, 1000 U/ml) or with a combination of both up to 96 hours. Cells were harvested and total RNA was isolated. The changes in expression of mRNA coding for RANTES, the interleukines (IL)-6, 8, 10, 15, IFN-gamma, monocyte chemotactic protein-1 (MCP-1) and transforming growth factor-beta1 (TGF-beta1) during the stimulation were investigated using the ribonuclease protection assay.


RESULTS
IL-10 and IFN-gamma mRNA were detected in neither unstimulated nor stimulated cells. Human RPE cells constitutively express the mRNA for IL-6, MCP-1, IL-8, IL-15, TGF-beta1 and, at very low levels, for RANTES. The TGF-beta1 mRNA expression was not influenced by either stimulation. The mRNA of the other factors was up-regulated for 24-48 h dependent on the stimulation.


CONCLUSIONS
Human RPE cells are able to increase their mRNA expression for the detected cytokines in response to the pro-inflammatory factors which are detectable in the rejection process. These up-regulated cytokines themselves are known to be involved in several inflammatory and immunological processes, suggesting their role in the rejection of transplanted RPE allografts.
Hollborn
M
M
Department of Ophthalmology, University of Leipzig, Leipzig, Germany. hollbm@server3.medizin.uni-leipzig.de
Enzmann
V
V
Barth
W
W
Wiedemann
P
P
Kohen
L
L
D002478:Cells, Cultured; D018925:Chemokines / Q000235:genetics*; D016207:Cytokines / Q000235:genetics*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D018925:Chemokines; D016207:Cytokines; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma