May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Stationary and Migrated Immunocompetent Cells in LPS–Induced Uveitis of GFP+–Chimeric Mice
Author Affiliations & Notes
  • M.R. Kuebbeler
    Dept. Vitreoretinal Surgery, Center of Ophthalmology, University of Cologne, Cologne, Germany
  • P.S. Muether
    Dept. Vitreoretinal Surgery, Center of Ophthalmology, University of Cologne, Cologne, Germany
    Center for Molecular Medicine (CMMC), University of Cologne, Cologne, Germany
  • I. Semkova
    Dept. Vitreoretinal Surgery, Center of Ophthalmology, University of Cologne, Cologne, Germany
  • M. Beyer
    Molecular Tumor Biology and Tumor Immunology, Clinic I for Internal Medicine, Cologne, Germany
  • N. Kociok
    Dept. Vitreoretinal Surgery, Center of Ophthalmology, University of Cologne, Cologne, Germany
  • A.M. Joussen
    Dept. Vitreoretinal Surgery, Center of Ophthalmology, University of Cologne, Cologne, Germany
    Center for Molecular Medicine (CMMC), University of Cologne, Cologne, Germany
  • Footnotes
    Commercial Relationships  M.R. Kuebbeler, None; P.S. Muether, None; I. Semkova, None; M. Beyer, None; N. Kociok, None; A.M. Joussen, None.
  • Footnotes
    Support  DFG JO 324 / 6–1 and ZMMK TV 76
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 2587. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      M.R. Kuebbeler, P.S. Muether, I. Semkova, M. Beyer, N. Kociok, A.M. Joussen; Stationary and Migrated Immunocompetent Cells in LPS–Induced Uveitis of GFP+–Chimeric Mice . Invest. Ophthalmol. Vis. Sci. 2006;47(13):2587.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : The generation of GFP+–chimeric mice allows evaluating ratios of residential and attracted cells in the retina. This study is to determine to which extent local retinal – and bloodstream derived immunocompetent cells participate in the inflammatory response associated with LPS–induced uveitis.

Methods: : C57/B6 mice were stably transplanted with whole bone marrow from b–actin promoted GFP+–heterozygous donors after lethal irradiation. Transformation rates were evaluated using FACS–analysis. Autoimmune–uveitis was induced via foodpad–injection of LPS. 24h after LPS injection, animals were enucleated and retinal flatmount preparation was conducted. Transcardial perfusion with concanavalin A was performed for differentiation of extravasal retinal cells. Extravasal cells were counted via an image analysis software. For colocalization and differentiation studies, macrophages and dendritic cells were labelled with F4/80– and CD11c antibodies, respectively, and evaluated using fluorescence– and confocal microscopy.

Results: : FACS analysis of peripheral blood draws showed an average of 80% GFP+–transformation rate after bone marrow substitution. Perfused retinal flatmounts of non–uveitic control animals without uveitis were free of extravasal GFP–positive cells as well as dendritic cells confirmed by CD11c staining. Diffuse GFP–negative cells positive for F4/80 were detected. LPS–induced uveitis led to influx and extravasation of GFP–positive cells as well as an increase in GFP–negative cells positive for F4/80–macrophage and CD11c dendritic cell staining.

Conclusions: : The high amounts of GFP–negative cells positive for macrophage and dendritic cell staining in uveitic retinas was not explained the low number of GFP–negative cells transplanted. These findings suggest an activation of residential immunocompetent retinal cells together with an influx of bloodstream–derived cells.

Keywords: inflammation • uveitis-clinical/animal model 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×