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Sunao Sugita, Yuko Kawazoe, Ayano Imai, Yoshihiko Usui, Yoichiro Iwakura, Kikuo Isoda, Masataka Ito, Manabu Mochizuki; Mature Dendritic Cell Suppression by IL-1 Receptor Antagonist on Retinal Pigment Epithelium Cells. Invest. Ophthalmol. Vis. Sci. 2013;54(5):3240-3249. doi: 10.1167/iovs.12-11483.
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To determine whether retinal pigment epithelial (RPE) cells can inhibit mature dendritic cells (mDCs).
Cultured RPE cells were established from C57BL/6 mice. DCs were established from bone marrow cells of normal mice, and mDCc were induced by culture in medium containing granulocyte macrophage–colony-stimulating factor (GM-CSF) and IL-4 in the presence of lipopolysaccharide and TNF-α. Activation of mDCs was assessed by a proliferation assay and ELISA to measure the production of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-12p40). Expression of major histocompatibility complex (MHC) class II, CD11c, and costimulatory molecules such as CD80, CD86, programmed cell death 1 ligand 1 (PD-L1), and PD-L2 on mDCs or RPE-exposed mDCs was evaluated by immune staining and flow cytometry. Production of IL-1 receptor antagonist (IL-1Ra) by RPE cells was evaluated by oligonucleotide microarray or ELISA. Anti–IL-1Ra neutralizing antibodies or RPE cells from IL-1Ra knockout donors were used for the assay.
Cultured RPE cells greatly suppressed the activation of mDCs, especially the production of pro-inflammatory cytokines, and the expression of cell–surface molecules. Moreover, RPE cells significantly suppressed mixed lymphocyte reactions by mDCs. In an examination of immunoregulatory candidate molecules, RPE cells expressed much higher levels of IL-1Ra as compared with control cells, and RPE cells pretreated with recombinant TNF-α and/or IL-1β produced high levels of IL-1Ra. RPE cells in the presence of anti–IL-1Ra antibodies, but not other candidate factors, failed to suppress activation by mDCs. In addition, RPE cells from IL-1Ra null donors failed to suppress mDC activation.
Our results suggest that ocular resident cells can produce pro-inflammatory cytokine antagonist that suppresses antigen-presenting cell activation.
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