title: Monocyte/macrophages promote vasculogenesis in choroidal neovascularization in mice by stimulating SDF-1 expression in RPE cells.
abstract: BACKGROUND
  Monocyte-macrophages play important roles in choroidal neovascularization (CNV); however, the mechanism is unclear. This study investigated the effects of monocyte depletion on laser-induced CNV in mice, especially the involvement of bone marrow-derived cells (BMCs) and underlying molecular mechanisms.
  
  
  METHODS
  Clodronate-liposomes (lip) were used to deplete monocytes and their effect on retinal pigmental epithelium (RPE) cells, endothelial cells, and BMCs was analyzed. Green fluorescent protein (GFP)-chimeric mice were developed by transplanting bone marrow cells from GFP transgenic mice to C57BL/6 J mice. CNV was induced by laser photocoagulation. Chimeric mice were intravenously treated with clodronate-lip, PBS-lip or PBS, 1 day before and after lasering. Histopathological and choroidal flatmount analysis were performed to measure CNV severity and BMCs recruitment. BMCs expression of endothelial cell marker CD31 and vascular smooth muscle cell marker α-SMA in CNV were detected by immunofluorescence. Expression of stromal cell-derived factor-1 (SDF-1) protein in vivo was detected by immunofluorescence as well as ELISA assay. SDF-1 was also examined by RT-PCR and ELISA in a human monocytes-RPE cells co-culturing system.
  
  
  RESULTS
  No valid evidence for the toxicity of clodronate-lip was found. Depletion led to significant inhibition of CNV and BMCs recruitment into laser spots on days 3 and 14, reduced BMC expression of CD31 and α-SMA on day 14, and decreased expression of SDF-1 in vivo on day 3. SDF-1 was mostly within and around the RPE cells in the laser lesion. SDF-1 was dramatically up-regulated in RPE cells after co-culturing with monocytes.
  
  
  CONCLUSIONS
  Monocytes may promote experimental CNV, especially BMC contribution in mice, by promoting SDF-1 production in RPE cells.
authors.affiliation: Department of Ophthalmology, Eye Institute of Chinese PLA, Xijing Hospital, Fourth Military Medical University, 710032, Shaanxi Province, China.
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Shi
authors.forename: Yu-sheng
authors.initials: YS
authors.lastname: Wang
authors.forename: Zhao-xia
authors.initials: ZX
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Cai
authors.forename: Jing
authors.initials: J
authors.lastname: Zhou
authors.forename: Hui-yuan
authors.initials: HY
authors.lastname: Hou
authors.forename: Nico
authors.initials: N
authors.lastname: van Rooijen
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism; D016026:Bone Marrow Transplantation; D054377:Chemokine CXCL12 / Q000235:genetics / Q000378:metabolism*; D002678:Chimera; D002829:Choroid / Q000098:blood supply*; D020256:Choroidal Neovascularization / Q000378:metabolism* / Q000473:pathology; D004002:Clodronic Acid / Q000633:toxicity; D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D049452:Green Fluorescent Proteins / Q000235:genetics; D047589:Leukocyte Reduction Procedures; D008264:Macrophages / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009000:Monocytes / Q000502:physiology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D000199:Actins; D054377:Chemokine CXCL12; C515256:Cxcl12 protein, mouse; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D012333:RNA, Messenger; C544013:alpha-smooth muscle actin, mouse; D004002:Clodronic Acid; D049452:Green Fluorescent Proteins