July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Detection of complement activators in immune attack eyes after iPS-derived retinal pigment epithelial cell transplantation
Author Affiliations & Notes
  • Sunao Sugita
    Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan
  • Kenichi Makabe
    Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan
  • Shota Fujii
    Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan
  • Masayo Takahashi
    Laboratory for Retinal Regeneration, Center for Developmental Biology, RIKEN, Kobe, Japan
  • Footnotes
    Commercial Relationships   Sunao Sugita, None; Kenichi Makabe, None; Shota Fujii, None; Masayo Takahashi, None
  • Footnotes
    Support  Research Center Network for Realization of Regenerative Medicine from the Japan Agency for Medical Research and Development, AMED
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 4998. doi:
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    • Get Citation

      Sunao Sugita, Kenichi Makabe, Shota Fujii, Masayo Takahashi; Detection of complement activators in immune attack eyes after iPS-derived retinal pigment epithelial cell transplantation. Invest. Ophthalmol. Vis. Sci. 2018;59(9):4998.

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

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Abstract

Purpose : To determine whether human induced pluripotent stem (iPS) cell-derived retinal pigment epithelial (RPE) cells (iPS-RPE) can express complement factors.

Methods : To confirm expression of complement factors in human iPS-RPE cells, flow cytometry, immunohistochemistry (IHC), ELISA, and quantitative RT-PCR (qRT-PCR) were performed for the following: C3, C5, CFB (Factor B), C5b-9 (membrane attack complex: MAC), CFH (Factor H), CFI (Factor I), CD46, CD55, CD59, clusterin, and vitronectin. iPS-RPE cells in the presence of recombinant IFN-g, recombinant TNF-a, lipopolysaccharide, supernatants of naïve T cells, and T helper 1 (Th1) cells were also prepared. For the transplantation, after preparation of iPS-RPE cells from cynomolgus monkeys, the iPS-RPE cells (allografts) were transplanted into the subretinal space in monkeys. After surgery, transplanted monkeys were sacrificed for IHC evaluation of the retinal section and determination of complement factors (C3, C5, CFB, MAC, and C1q), cytokines (IFN-gamma), and immunoglobulin G (IgG).

Results : Human iPS-RPE cells expressed complement activators and inhibitors. iPS-RPE cells highly expressed complement factors during inflammatory conditions, especially IFN-g exposure including Th1 cell supernatants. In immune attack eyes after allogeneic iPS-RPE cell transplantation, complement activators such as C3, CFB, C5, and membrane attack complex (MAC) were detected around the host RPE layer, grafted RPE cells, inflammatory retinal lesions, and transplanted subretinal space. In addition, we observed a large number of C1q and IgG double positive and IFN-gamma positive inflammatory cells in the retinal sections.

Conclusions : iPS-derived RPE cells greatly expressed complement factors. Thus, RPE cells might be activated and produce complement factors after exposure to infiltrating inflammatory cells in the eye.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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