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AbstractDO.05.07 Effect of complement and zymosan on retinal pigment epithelial cells Lück K.1, Wasmuth S.1, Hennig M.1, Pauleikhoff D.2
1Ophtha-Lab of Department of Ophthalmology at St. Franziskus Hospital, Muenster; 2Department of Ophthalmology at St. Franziskus Hospital, Muenster Objective: A mutation in complement factor H is often found in patients with age-related macular degeneration (AMD). Thus exceeding amounts of complement can be activated. Therefore, the influence of artificially activated complement on the production of cytokines relevant in AMD by retinal pigment epithelial (RPE) cells was examined in this study.
Methods: Adherent RPE cells were cultured with different concentrations of native and heat-inactivated serum. Further, the RPE cells were treated with zymosan, a complement activating constituent of the yeast cell wall. Serum and zymosan in combination was also tested. After 24 hours the amount of interleukin (IL)-6, -8 and vascular endothelial growth factor (VEGF) in the supernatants were examined by sandwich ELISA. Cells incubated in serum free medium were used to evaluate the baseline production.
Results: Untreated RPE cells produced only low levels of IL-6, -8 and VEGF. The addition of serum stimulated RPE cells to a concentration-dependent release of the examined cytokines and chemokines. Zymosan intensified the production of IL-6 and -8, but did not impair the synthesis of VEGF. Native serum clearly increased the content of IL-6 and -8 compared to heat inactivated serum. The combined treatment of RPE cells with serum and zymosan resulted in a more than additive enhancement of IL-6 release. The content of IL-8 and VEGF was additive.
Conclusions: The release of IL-8 and VEGF by RPE cells cultivated with serum and zymosan was additive. In contrast, the production of IL-6 was strongly increased due to the activation of complement within the serum by zymosan. Thus, the activation of complement could stimulate inflammatory processes by activated RPE cells leading to AMD.
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