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Takahiro Yamawaki, Eiko Ito, Jun Yamada, Shigeru Kinoshita, Chie Sotozono, Junji Hamuro; Investigation of the interactions between macrophages and retinal pigment epithelium (RPE) cells in AMD. Invest. Ophthalmol. Vis. Sci. 2017;58(8):5737.
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© ARVO (1962-2015); The Authors (2016-present)
Age-relate macular degeneration (AMD) is partly caused by chronic inflammation. The purpose of this study is to clarify the interactions between macrophages(MPs) and RPE cells in coculture systems.
Adherent peritoneal cells or murine MPs cell line Raw 264.7 was cocultured with primary RPE cells taken from C57BL/6 mice. MCP-1, IL-6, VEGF, and TNF-α in the culture supernatants(CSs) were quantified by ELISA. The expression profiles, in cocultures, of complement-associated genes, TNF-α, and angiogenesis-associated genes were analyzed by quantitative real-time PCR. To investigate the effect of MPs polarization, MPs cultured with LPS and IFNγ or IL4 were also used.
The production of MCP-1, IL-6, and VEGF were synergistically elevated when primary MPs or RAW264.7 and RPE cells were co-cultured compared with those derived from sole cultures of MPs and RPE cells. TNFα production by MPs was suppressed by RPE cells. Coculture of RPE cells with RAW264.7 cells increased the gene expression of C3, CFB, and VEGF genes, whereas it reduced those of complement regulatory factors CFH, CD59, clusterin, TNFα, and PEDF. The synergistic effect was more increased when MPs were polarized into M1 compared with when polarized into M2.
Our findings indicate the presence of ingenious interactions between MPs and RPE cells that forces the inflammation and complement activation in the vicinity of RPE cells, and the interactions were more prominent when MPs were polarized into M1.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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