July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Bacterial infection promotes spontaneous choroidal neovascularization (sCNV) development in the JR5558 mouse
Author Affiliations & Notes
  • YU SU
    Schepens Eye Research Insitute of Massachusess Eye and Ear, Harvard Medical School Department of Ophthalmology, Malden, Massachusetts, United States
    Ophthalmic Center, Renmin Hospital of Wuhan University, Wuhan, Hubei, China
  • Franco Aparecido Rossato
    Schepens Eye Research Insitute of Massachusess Eye and Ear, Harvard Medical School Department of Ophthalmology, Malden, Massachusetts, United States
  • Yin Shan Eric Ng
    Schepens Eye Research Insitute of Massachusess Eye and Ear, Harvard Medical School Department of Ophthalmology, Malden, Massachusetts, United States
  • Footnotes
    Commercial Relationships   YU SU, None; Franco Aparecido Rossato, None; Yin Shan Eric Ng, None
  • Footnotes
    Support  This project is supported by Grimshaw Foundation AMD Research Grant from Mass Eye and Ear and Bright Focus Foundation AMD Research grant. Yu Su is supported from China Scholarship Council (Grant No. 201606275174) and National Natural Science Foundation of China (Grant No. 81500744)
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4902. doi:https://doi.org/
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      YU SU, Franco Aparecido Rossato, Yin Shan Eric Ng; Bacterial infection promotes spontaneous choroidal neovascularization (sCNV) development in the JR5558 mouse. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4902. doi: https://doi.org/.

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

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Abstract

Purpose : We hypothesized that bacterial infection induces sub-clinical inflammation of the retinal pigment epithelium (RPE) via the activation of the Toll-like receptor 2 (TLR2) and contributes to sCNV pathogenesis. We aim at determining the effect of short-term systemic antibiotic on sCNV development and the mechanism of TLR2-mediated RPE inflammation.

Methods : Two different breeding groups of JR5558 mice were set up from the same litter (F0), with and without Ditrim (0.96 mg/ml) in drinking (F1 mice), and for different duration (F2-A and F2-B mice) (Fig 1A). FITC-conjugated isolectin B4 and anti-CD45 staining were used to quantify sCNV and immune cells, respectively. NF-κB expression was determined by staining. RPE/choroid expression of VEGFA, TLR2, VE-cadherin, CD45, MCP-1, IL-6 was determined by qPCR. RPE/choroid/sclera explants from C57B6J mice were cultured with and without Pam2+CEP (carboxyethyl-pyrrole-dipeptide) on the transwell, and expression of VEGFA, TLR2, VE-cadherin, CD45, IL-6 was determined by qPCR.

Results : In the F1 eyecups both the number of sCNV (13±9 vs. 32±15, P<0.0001) and CNV area per eye (21076±14687 vs. 70210±49389 pixel area, P<0.0001) were significantly reduced in the Ditrim group compared to control. In the F2-A group, in which Ditrim was stopped at P0, the number of CNV (9 ± 2 vs. 32±15, P<0.0001) and CNV area per eye (12048 ± 3161 vs. 70210±49389 pixel area, P<0.0001) were significantly reduced, and the numbers of CD45+ leukocytes in both the F2-A and F2-B groups were significantly reduced compared to the control (93±37, 62±20, vs. 284±53, P<0.001 for both) (Fig 1B). NF-κB staining was lower in all the F2 groups compared to control. Expression of TLR2, VEGFA, IL-6 by RPE/choroid was significantly decreased in all the Ditrim treated F1 mice compared to control (P<0.05 for all). For the explants, Pam2+CEP treatment significantly increased the expression of VE-cadherin, VEGF,TLR2 and MCP-1 compared to controls (P<0.03 for all).

Conclusions : Systemic Ditrim inhibits sCNV and suppresses inflammation of the RPE/choroid indirectly, which is likely mediated by the anti-bacterial activity of Ditrim. Furthermore, activation of TLR2 by bacterial ligand induces robust pro-inflammatory cytokines and angiogenic marker expression by RPE/choroid/sclera explants and drives sCNV development.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

Fig1 (A) Animal experiment design.(B) Quantification of sCNV and inflammation.

Fig1 (A) Animal experiment design.(B) Quantification of sCNV and inflammation.

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