Skip to main content
. 2020 May 5;9:e54257. doi: 10.7554/eLife.54257

Figure 2. Requirements of IL-4/IL-4Rα in the inductive phase of choroidal neovascularization (CNV).

(a) Effect of systemic administration of recombinant murine IL-4 (rIL-4) or recombinant IL-13 (rIL-13) in the inductive phase in a CNV model. (n = 7–19 eyes/group). (b) Inhibitory effect of systemic administration of anti-IL-4 antibody in the inductive phase of CNV formation. (n = 7–20 eyes/group). (c) Impaired CNV development in Il4 deficient mice. CNV development is significantly impaired in Il4-/- mice compared to wild type. (n = 8–9 eyes/group). (d) Impaired CNV development by IL-4 receptors deficiency. CNV development is significantly impaired in Il4ra-/- and Il4ra+/- mice compared to wild type. This impairment is more marked in the homozygotes. CNV development is not impaired for Il13ra1-/- mice. (n = 7–17 eyes/group) (e) Bone marrow chimeric mice reconstituted with GFP transgenic bone marrow cells that were exposed to laser to induce CNVs. The CNV lesions after 14 days were analyzed for lineage cell markers by immunohistochemistry. CNVs are formed as clusters of isolectin IB4-positive vascular endothelial cells (red). Bone marrow-derived cells (green) were co-localized with isolectin-positive vascular endothelial cells. IL-4 positive cells (yellow) are distributed at the margins of the CNVs and precisely match the location of the bone marrow-derived cells (green). IL-4Rα-positive cells (cyan) partly overlapped the bone marrow-derived cells, and precisely match the location of the vascular endothelial cells in the CNV lesion. *p<0.005, **p<0.001, ***p<0.0005. Nested ANOVA with post hoc test. Scale 10 μm.

Figure 2—source data 1. Requirements of IL-4/IL-4Rα in the inductive phase of CNV.

Figure 2.

Figure 2—figure supplement 1. Kinetics of IL-4, IL-4Rα, CCR2 and CD11b-expressing cells and GFP-positive bone marrow derived cells determined by immunohistochemical analyses.

Figure 2—figure supplement 1.

The distribution of GFP transgenic bone marrow cells (green) shows dynamic changes after laser irradiation. GFP transgenic bone marrow cells remain around the choroidal scar at 1 day after the laser irradiation. Then GFP transgenic bone marrow cells spread out in the subretinal space at 3 days after laser irradiation and some GFP transgenic bone marrow cells return to the center area of CNV lesion. IL-4 (yellow), IL-4Rα-, CCR2-, and CD11b-positive cells (cyan) partly overlap the distribution of the GFP transgenic bone marrow cells. Scale 50 μm.
Figure 2—video 1. Localization of bone marrow-derived cells and endothelial cells in the retinal tissue of GFP bone marrow chimeric mouse in a 3D rendering 7 days after laser irradiation.
Download video file (970.5KB, mp4)
CCR2 (cyan) expressing bone marrow-derived cells (green) are incorporated into CD31 (red) expressing vascular endothelial morphology (arrows) and were spatially distinct from monocytes lineage cells. Scale 50 μm.