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In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy
Published on: November 11, 2017
Experimental autoimmune uveoretinitis (EAU)-related tissue damage and angiogenesis is reduced in CCL2⁻/⁻CX₃CR1gfp/gfp
Jiawu Zhao1, Mei Chen1, Heping Xu1
1Centre for Experimental Medicine, School of Medicine, Dentistry and Biomedical Sciences, Queen's University Belfast, United Kingdom.
Purpose:
To investigate the roles of the CCL2-CCR2 and CX₃CL1-CX₃CR1 pathways in experimental autoimmune uveoretinitis (EAU)-mediated retinal tissue damage and angiogenesis.
Methods:
The C57BL/6J wild-type (WT) and CCL2(-/-)CX₃CR1(gfp/gfp) (double knockout [DKO]) mice were immunized with IRBP₁₋₂₀. Retinal inflammation and tissue damage were evaluated clinically and histologically at different days postimmunization (p.i.). Retinal neovascular membranes were evaluated by confocal microscopy of retinal flat mounts, and immune cell infiltration by flow cytometry.
Results:
At day 25 p.i., DKO mice had lower clinical and histological scores and fewer CD45(high)CD11b(+) infiltrating cells compared with WT mice. The F4/80(+) macrophages constitute 40% and 21% and CD11b(+)Gr-1(+)Ly6G(+) neutrophils constitute 10% and 22% of retinal infiltrating cells in WT and DKO mice, respectively. At the late stages of EAU (day 60-90 p.i.), DKO and WT mice had similar levels of inflammatory score. However, less structural damage and reduced angiogenesis were detected in DKO mice. Neutrophils were rarely detected in the inflamed retina in both WT and DKO mice. Macrophages and myeloid-derived suppressor cells (MDSCs) accounted for 8% and 3% in DKO EAU retina, and 19% and 10% in WT EAU retina; 71% of infiltrating cells were T/B-lymphocytes in DKO EAU retina and 50% in WT EAU retina.
Conclusions:
Experimental autoimmune uveoretinitis-mediated retinal tissue damage and angiogenesis is reduced in CCL2(-/-)CX₃CR1(gfp/gfp) mice. Retinal inflammation is dominated by neutrophils at the acute stage and lymphocytes at the chronic stage in these mice. Our results suggest that CCR2(+) and CX₃CR1(+) monocytes are both involved in tissue damage and angiogenesis in EAU.
Insights
Experimental autoimmune uveitis causes retinal damage and new blood vessel growth. Blocking the CCL2-CCR2 and CX₃CL1-CX₃CR1 pathways in mice reduced this damage and angiogenesis, suggesting their role in the disease process.
Area of Science:
- Immunology
- Ophthalmology
- Pathology
Background:
- Experimental autoimmune uveoretinitis (EAU) is an immune-mediated disease causing retinal inflammation and damage.
- The chemokine pathways CCL2-CCR2 and CX₃CL1-CX₃CR1 are implicated in immune cell recruitment and inflammation.
- Understanding these pathways is crucial for developing targeted therapies for uveitis.
Purpose of the Study:
- To investigate the roles of the CCL2-CCR2 and CX₃CL1-CX₃CR1 pathways in EAU-induced retinal tissue damage and angiogenesis.
- To determine the impact of combined CCL2 and CX₃CR1 deficiency on immune cell infiltration and disease progression in EAU.
Main Methods:
- C57BL/6J wild-type (WT) and CCL2(-/-)CX₃CR1(gfp/gfp) (double knockout [DKO]) mice were immunized to induce EAU.
- Retinal inflammation, tissue damage, and neovascularization were assessed clinically and histologically.
- Immune cell infiltration was quantified using flow cytometry.
Main Results:
- DKO mice exhibited reduced clinical and histological scores of EAU compared to WT mice at day 25 post-immunization.
- Macrophages and lymphocytes were the predominant infiltrating immune cells in DKO mice during chronic EAU, while neutrophils were prominent in acute EAU.
- DKO mice showed significantly less structural damage and reduced angiogenesis at later stages of EAU (day 60-90 post-immunization).
Conclusions:
- The combined deficiency of CCL2 and CX₃CR1 pathways significantly reduces retinal tissue damage and angiogenesis in experimental autoimmune uveitis.
- Monocytes expressing CCR2 and CX₃CR1 play critical roles in mediating both tissue damage and neovascularization in EAU.
- Targeting these chemokine pathways may offer a therapeutic strategy for managing uveitis and its complications.
