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The retinal pigment epithelium (RPE) induces FasL and reduces iNOS and Cox2 in primary monocytes

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Abstract

Purpose

Retinal pigment epithelium (RPE) cells may alter the phenotype of monocytes by soluble factors that may be influenced by stimulation of the RPE. Since RPE cells carry the toll-like receptor-3 (TLR3) that detects and reacts to viral infection through binding of dsRNA we investigated the effects of RPE cells with or without TLR3 stimulation on blood-derived monocytes with respect to regulation of pro-/anti-inflammatory cytokines, anti-angiogenic factors and migratory properties.

Methods

Primary RPE cells were prepared from porcine eyes; monocytes were prepared from porcine blood. TLR3 activation was induced by polyinosinic:polycytidylic acid (Poly I:C). RPE cells were stimulated with Poly I:C in different concentrations for 24 hours and a cell culture supernatant was applied to the monocytes. Expression of CD14 and Fas ligand (FasL) was determined via flow cytometry. The expression of IL-6, IL-1ß, TNFα, Cox2, iNOS and IL-10 was determined via quantitative RT-PCR. Migration was determined using Boyden chamber experiments.

Results

The supernatant of RPE cells, irrespective of TLR3 activation, induced FasL expression in the monocytes. Expression of iNOS and Cox2 was reduced by RPE cells and the reduction of Cox2 but not if iNOS was lost under TLR3 activation. No induction of IL-6, IL-1ß, IL-10 or TNFα by the RPE was seen. TLR3-activated RPE cells induced monocyte migration.

Conclusion

RPE cells induce an upregulation of FasL and a downregulation of iNOS and Cox2 without upregulating inflammatory cytokines, possibly inducing an anti-angiogenic phenotype in the monocytes. This phenotype is still upheld after challenging RPE cells with dsRNA, mimicking a viral infection.

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Acknowledgments

This project was supported by the German Ophthalmological Society. Data of this study has been presented at the ARVO meeting 2013. No conflict of interest exists regarding this study. Independently of this study, AK has received research funding and lecture fees and has acted as a consultant for Novartis Pharma.

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Authors and Affiliations

  1. Department of Ophthalmology, University of Kiel, University Medical Center, Arnold-Heller-Str. 3, 24105, Kiel, Germany

    Christin Hettich, Sebastian Wilker, Johann Roider & Alexa Klettner

  2. Department of Anatomy, University of Kiel, Kiel, Germany

    Rolf Mentlein & Ralph Lucius

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  1. Christin Hettich
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  2. Sebastian Wilker
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  3. Rolf Mentlein
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  4. Ralph Lucius
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  5. Johann Roider
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  6. Alexa Klettner
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Correspondence to Alexa Klettner.

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Hettich, C., Wilker, S., Mentlein, R. et al. The retinal pigment epithelium (RPE) induces FasL and reduces iNOS and Cox2 in primary monocytes. Graefes Arch Clin Exp Ophthalmol 252, 1747–1754 (2014). https://doi.org/10.1007/s00417-014-2742-z

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  • DOI: https://doi.org/10.1007/s00417-014-2742-z

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