Open AccessChoroidal Neovascularization Is Inhibited in Splenic-Denervated or Splenectomized Mice with a Concomitant Decrease in Intraocular Macrophage
Author(s) -
Xue Tan,
Katsuhito Fujiu,
Ichiro Manabe,
Junko Nishida,
Reiko Yamagishi,
Yuya Terashima,
Kouji Matsushima,
Toshikatsu Kaburaki,
Ryozo Nagai,
Yasuo Yanagi
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0160985
Subject(s) - choroidal neovascularization , lesion , pathology , spleen , cd8 , neovascularization , medicine , macrophage , immunology , biology , ophthalmology , angiogenesis , immune system , macular degeneration , in vitro , biochemistry
Purpose To determine the involvement of sympathetic activity in choroidal neovascularization (CNV) using laser-induced CNV in a mouse model. Methods We investigated changes in the proportions of intraocular lymphocytes, granulocytes, and three macrophage subtypes (Ly6C hi , Ly6C int , and Ly6C lo ) after laser injury in mice using flow cytometry, and evaluated CNV lesion size in mice lacking inflammatory cells. Further, we evaluated the lesion size in mice administered the β3 receptor antagonist, splenic-denervated and splenectomized mice. We also assessed changes in the proportions of intraocular macrophages and peripheral blood monocytes in splenic-denervated and splenectomized mice. Lastly, lesion size was compared between splenic-denervated mice with or without adoptive transfer of macrophages following laser injury. After Ly5.1 mice spleen-derived Ly6C hi cells were transferred into Ly5.2 mice, the proportions of intraocular Ly5.1 + Ly6C hi cells were compared. Results In WT mice, the proportion of CD4 + T cells recruited into the eye increased progressively from day 3 to day 7 after laser injury, whereas, intraocular CD8 + T cells did not change significantly. Proportions of B220 + cells, granulocytes, and two subtypes of intraocular macrophages (Ly6C hi and Ly6C lo ) peaked at day 3 following laser injury. In contrast, Ly6C int/lo CD64 + subtype showed a significantly higher percentage at day 7 after laser injury. There were no differences in lesion size between CD4 –/– or Rag2 –/– mice and controls, whereas lesion size was significantly reduced in CCR2 −/− mice and clodronate liposome-treated mice. CNV lesion area was significantly reduced in mice with β3 blocker treatment, splenic-denervated and splenectomized mice compared with controls. Intraocular Ly6C hi macrophages were also reduced by splenic denervation or splenectomy. Adoptive transfer of spleen-derived Ly6C hi cells increased the lesion size in splenic-denervated mice. Compared with controls, intraocular donor-derived Ly6C hi cells recruited into the eye were reduced in splenic-denervated and splenectomized mice. Conclusions Although lymphocytes had little effect on CNV formation, Ly6C hi macrophages/monocytes exacerbated CNV in mice. Sympathetic activity might contribute to CNV via the recruitment of macrophages to the eye.