gms | German Medical Science

25th Annual Meeting of the German Retina Society

German Retina Society

01.06. - 02.06.2012, Münster

The role of VEGF-expressing mononuclear phagocytes in a murine model of laser-induced choroidal neovascuarization

Meeting Abstract

  • Torsten A. Urzynicok - IMMEI, Rheinische Friedrich Wilhelms Universität, Bonn
  • A.F. Alex - Universitäts-Augenklinik Münster
  • D. Engel - IMMEI, Rheinische Friedrich Wilhelms Universität, Bonn
  • C. Kurts - IMMEI, Rheinische Friedrich Wilhelms Universität, Bonn
  • N. Eter - Universitäts-Augenklinik Münster

German Retina Society. 25th Annual Conference of the German Retina Society. Münster, 01.-02.06.2012. Düsseldorf: German Medical Science GMS Publishing House; 2012. Doc12rg07

DOI: 10.3205/12rg07, URN: urn:nbn:de:0183-12rg079

This is the translated version of the article.
The original version can be found at: http://www.egms.de/de/meetings/rg2012/12rg07.shtml

Published: May 30, 2012

© 2012 Urzynicok et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.


Outline

Text

Background: The aim of this study was to evaluate, whether infiltrating or resident phagocytes are a source of VEGF that causes neovascularization in a murine CNV-model.

Methods: For visualization of mononuclear phagocytes CX3CR1-reporter mice were used.

These mice express GFP controlled by the CX3CR1-promoter, which is active in mononuclear phagocytes (dendritic cells, macrophages, microglia). In CX3CR1-reportermice with or without

CCR2-expression 10 laserspots were set (50 µm, 200 mW and 0.1 s duration) using a slitlamp applicator. The retinal pigment epithelium and Bruch“s membrane were ruptured. 3 and 6 days post laser eyes was enucleated, retina and choroid of both eyes collected, singlecellsuspensions prepared and stained with fluorescence-labeled antibodies for different celltypes (CD11b, CX3CR1, CD45, CD31) for analyzing VEGF-production by flow cytometry. CNV-areas were measured in choroidal flatmounts, while mononuclear phagocytes were localized in retinal flatmounts.

Results: The numbers of VEGF-producing mononuclear phagocytes (CD11b+) increased significantly 3 days post laser, especially those of retinal microglia (CX3CR1high CD45low) and choroidal macrophages (CX3CR1low CD45high). Microscopy revealed that these cells accumulated colocalized to laserspots. The number of VEGF-expressing choroidal macrophages increased on day 3 more than 28-fold and declined until day 6. The dramatic increase was absent in CCR2-deficient mice. In contrast the increase of VEGF-producing retinal microglia was CCR2-independent. CNV-area was diminished in flatmounts of CCR2-deficient mice.

Conclusions: VEGF-production of ocular phagocytes is quantifiable by flow cytometry.

CCR2-dependent macrophages are a relevant source of VEGF and accumulate post lasertreatment. CCR2-inhibition reduces CNV and may represent a therapeutical approach for an antineovascular therapy.