June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
CCR2-dependent recruitment of VEGF-producing macrophages promotes neovascularization after laser injury
Author Affiliations & Notes
  • Torsten Andreas Krause
    University Clinic of Bonn, Institute of Experimental Immunology, Bonn, Germany
  • Anne Alex
    University of Muenster, Dept. of Ophthalmology, Muenster, Germany
  • Daniel Engel
    University Clinic of Bonn, Institute of Experimental Immunology, Bonn, Germany
  • Christian Kurts
    University Clinic of Bonn, Institute of Experimental Immunology, Bonn, Germany
  • Nicole Eter
    University of Muenster, Dept. of Ophthalmology, Muenster, Germany
  • Footnotes
    Commercial Relationships Torsten Andreas Krause, None; Anne Alex, None; Daniel Engel, None; Christian Kurts, None; Nicole Eter, Novartis (F), Bayer (R), Heidelberg Engineering (R), Sanofi Aventis (C), Allergan (C), Bausch and Lomb (C)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 328. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Torsten Andreas Krause, Anne Alex, Daniel Engel, Christian Kurts, Nicole Eter; CCR2-dependent recruitment of VEGF-producing macrophages promotes neovascularization after laser injury. Invest. Ophthalmol. Vis. Sci. 2013;54(15):328. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: Age-related macular degeneration (AMD) is the most prevalent cause of blindness in the elderly, and its exudative subtype critically depends on local production of vascular endothelial growth factor (VEGF). In tumour models, mononuclear phagocytes (MPh) produce VEGF. MPh including dendritic cells, macrophages and microglia cells, express CX3CR1. MPh precursors, such as monocytes, can be recruited to sites of inflammation by CCR2. Here we studied VEGF production by MPh in a laser-induced murine model of choroidal neovascularisation (CNV) that mimics CNV in exudative AMD, and the impact of CCR2 on such production.

Methods: CCR2-competent and -deficient CX3CR1-reporter mice expressing GFP in MPh were used to visualize these cells by 3 complementary fluorescence-based techniques. CNV was induced by laser injury rupturing Bruch's membrane beneath the retina. 3 and 6 days later, eyes were enucleated to prepare single cell suspensions from the retina and the choroid. Cellular subsets were identified and their VEGF-production quantified by flow cytometry. Scanning-Laser-Ophthalmoscopy (SLO) in the 488 nm autofluorescence mode was used to localize MPh in living animals. CNV areas were measured in choroidal flatmounts by isolectin staining.

Results: VEGF-positive phagocytes increased 3 days post laser, with microglia being dominant in the retina (2fold) and macrophages in the choroid (>25fold). On day 6, VEGF-expressing microglia numbers remained increased, whereas macrophages had already declined. This temporary increase in macrophage numbers was abrogated in the absence of CCR2, whereas microglia was CCR2-independent. These macrophages were the only source of VEGF that upregulated their VEGF-production at day 3, but not later. Consistent with flow-cytometry, microscopy and in vivo SLO revealed that Mph accumulated in CNV areas. This area was reduced in CCR2-deficient mice on day 14, but not on day 21.

Conclusions: We established an intracellular staining method for VEGF-production and describe a non-invasive in vivo imaging method for ocular MPh. These MPh were identified as relevant producers of VEGF as they accumulate responding to laser injury. The immune cell related source of VEGF decreases by an impaired recruitment to the choroid. Infiltrating CCR2-dependent macrophages specifically increase CNV, indicating that CCR2 inhibition might be a temporal strategy to prevent CNV in AMD.

Keywords: 453 choroid: neovascularization • 529 flow cytometry • 557 inflammation  
×
×

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.

×