June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Retinal pigment epithelial (RPE) cells convert bone marrow derived macrophages into myeloid-derived suppressor cells with a novel phenotype
Author Affiliations & Notes
  • Heping Xu
    Centre for Vision and Vascular Science, Queen's University Belfast, Belfast, United Kingdom
  • Chang Luo
    Centre for Vision and Vascular Science, Queen's University Belfast, Belfast, United Kingdom
  • Mei Chen
    Centre for Vision and Vascular Science, Queen's University Belfast, Belfast, United Kingdom
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 144. doi:
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      Heping Xu, Chang Luo, Mei Chen; Retinal pigment epithelial (RPE) cells convert bone marrow derived macrophages into myeloid-derived suppressor cells with a novel phenotype. Invest. Ophthalmol. Vis. Sci. 2013;54(15):144.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: We have shown previously that macrophages/microglia accumulate in the subretinal space in the aging eye. The phenotype and function of subretinal macrophages /microglia remain elusive. Subretinal macrophages are in close contact with retinal pigment epithelial (RPE) cells, which may affect / regulate macrophage functions. The aim of this study was to investigate the effect of RPE cells on the phenotype and function of bone marrow-derived macrophages (BM-DMs).

Methods: Bone marrow cells from C57BL/6 mice were cultured in DMEM supplemented with GM-CSF for 5 day to generate BM-DMs. The phenotype of BM-DMs was confirmed by flow cytometry as CD11b+F4/80+MHC-II+Gr1-. Primary RPE cells were cultured from C57BL/6 mice and confirmed by RPE65 and cytokeratin staining. BM-DMs were co-cultured with sub-confluent RPE cells for different times. Macrophages were then collected for phenotypic and functional assays.

Results: Co-culture of BM-DMs with RPE cells results in a time-dependent down-regulation of MHC-II expression and the generation of CD11b+F4/80+Gr1+ myeloid-derived suppressor cells (MDSC). Real-time RT-PCR analysis showed that RPE-induced MDSCs expressed high levels of IL-6, IL-1β, and Arginase-1, but lower levels of IL-12p40 and TNF-α compared to naïve BM-DMs. The expression levels of iNOS, TGF-β and Ym1 did not differ between native BM-DMs and RPE-induced MDSCs. Furthermore, a functional study showed that these MDSCs could suppress anti-CD3 antibody mediated T cell activation and proliferation.

Conclusions: Our results suggest that RPE cells can convert bone-marrow derived macrophages into myeloid-derived suppressor cells under in vitro culture conditions. RPE-induced myeloid-derived suppressor cells are CD11b+F4/80+GR-1+MHC-IIlowIL-6+IL-1β+Arg-1+ and can suppress T cell activation.

Keywords: 557 inflammation • 701 retinal pigment epithelium • 555 immunomodulation/immunoregulation  
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