April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Origin of Retinal Dendritic Cells
Author Affiliations & Notes
  • U. Lehmann
    Ophthalmology, University of Minnesota, Minneapolis, Minnesota
  • N. D. Heuss
    Ophthalmology, University of Minnesota, Minneapolis, Minnesota
  • S. W. McPherson
    Ophthalmology, University of Minnesota, Minneapolis, Minnesota
  • D. S. Gregerson
    Ophthalmology, University of Minnesota, Minneapolis, Minnesota
  • Footnotes
    Commercial Relationships  U. Lehmann, None; N.D. Heuss, None; S.W. McPherson, None; D.S. Gregerson, None.
  • Footnotes
    Support  NIH Grant EY11542, NIH Grant EY16376, T32 EY07133
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 5926. doi:
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    • Get Citation

      U. Lehmann, N. D. Heuss, S. W. McPherson, D. S. Gregerson; Origin of Retinal Dendritic Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):5926.

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

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Abstract

Purpose: : Dendritic cells (DC) are fundamentally important in antigen presentation, but have been difficult to study for phenotype and function in retina. Because retinal CD11c+ cells change rapidly in response to minor injury, a partial optic nerve crush (ONC) was tested as a method to manipulate changes in retinal CD11c+ cells that would facilitate study of their frequency, morphology, origin, and function.

Methods: : An ONC was done to one eye of CD11c DTR-Tg mice, which express eGFP on the CD11c promoter. Chimeric B6CD11c-DTR Ly5.2 mice injected with B6Ly5.1 bone marrow and vice versa, were used to study the origin of retinal CD11c+ cells. CD11c-DTR mice received an optic nerve crush and were later depleted of their retinal CD11c+ cells by injecting diphtheria toxin (DTx) into their anterior chamber. In these sets of experiments, whole mounts were harvested at specified times and stained for DC and microglia (MG). Retinal CD11c+ cells were also analyzed for MHC-II expression. Whole retinas were flat-mounted and analyzed by fluorescence microscopy. Retinal blood vessels were stained with isolectin B4 to determine if MHC-II+ cells associated with them.

Results: : Partial ONC in one eye led to large increases of CD11c+ cells in the retinas of both eyes. Stained retina whole mounts showed MHC-II expression on CD11c+ cells and some were associated with blood vessels. Analysis of chimeric mice, in the absence of injury, suggested minimal turnover from the grafted stem cells after six months. Retinas from chimeric mice that received ONC 10 days prior to harvesting had many donor CD11c+ cells in the crushed eyes around the optic disc and in the periphery. Their contralateral retinas only had a few donor CD11c+ cells in the periphery. Retinas repeatedly depleted of CD11c+ cells by DTx injections also showed a reduction in the number of MG.

Conclusions: : The presence of retinal donor CD11c+ cells in B6Ly5.1 chimeric mice post-engraftment lead us to conclude that, following injury, these CD11c+ cells are derived from both MG, and that they are also recruited from the periphery. In the absence of injury, few of the CD11c+ cells were recruited from circulating precursors.

Keywords: antigen presentation/processing • immunohistochemistry • microglia 
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