title: The effect of podoplanin inhibition on lymphangiogenesis under pathological conditions.
abstract: OBJECTIVE
  Podoplanin has been shown to be a reliable marker of lymphatic endothelium, but its role in the lymphatic system has not been well investigated. The purpose of this study is to investigate the role of podoplanin in lymphangiogenesis and macrophage functions under inflammatory conditions.
  
  
  METHODS
  Mouse corneal suture and ear section models were used to induce lymphangiogenesis and macrophage infiltration. Antilymphatic vessel endothelial hyaluronan-1 (Anti-LYVE-1) antibody was used to visualize lymphatic vessels. Thioglycollate-induced macrophages (mps) were collected and cultured with lipopolysaccharide (LPS), IFN-γ, and anti-mouse podoplanin antibody (PMab-1). Podoplanin, NF-κB, and mitogen-activated protein kinase (MAPK) pathway expression were detected by Western blot analysis. The TNF-α secretion was measured by ELISA.
  
  
  RESULTS
  Administration of PMab-1, reduced lymphangiogenesis in the corneal suture and ear wound healing models. Also, PMab-1 suppressed mps infiltration at the site of wound healing. Moreover, administration of PMab-1 led to a significant suppression of the rejection reaction in the corneal transplantation model. Our in vitro experiments showed that PMab-1 suppressed TNF-α secretion from mps under inflamed conditions, especially secretion caused by LPS stimulation. We confirmed the effect of PMab-1 on mps under inflamed conditions with a Western blot experiment, which clearly showed that the phosphorylation signal of the MAPK and NF-κB pathways was suppressed by PMab-1.
  
  
  CONCLUSIONS
  Podoplanin neutralization resulted in inhibition of lymphatic growth associated with corneal and ear wound healing as well as mps inflammation. These data suggest that podoplanin is a novel therapeutic target for suppressing lymphangiogenesis and inflammation.
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Maruyama
authors.affiliation: Tohoku University Graduate School of Medicine, Department of Ophthalmology, Sendai, Japan.
authors.forename: Kazuichi
authors.initials: K
authors.lastname: Maruyama
authors.affiliation: Department of Regional Innovation, Tohoku University Graduate School of Medicine, Sendai, Japan.
authors.forename: Yukinari
authors.initials: Y
authors.lastname: Kato
authors.affiliation: Shiseido Innovative Science Research Center, Yokohama, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kajiya
authors.affiliation: Tohoku University Graduate School of Medicine, Department of Ophthalmology, Sendai, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Moritoh
authors.affiliation: Tohoku University Graduate School of Medicine, Department of Ophthalmology, Sendai, Japan.
authors.forename: Kotaro
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Shiseido Innovative Science Research Center, Yokohama, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Matsumoto
authors.affiliation: Shiseido Innovative Science Research Center, Yokohama, Japan.
authors.forename: Mika
authors.initials: M
authors.lastname: Sawane
authors.affiliation: Department of Pathology, Medical University Vienna, Vienna, Austria.
authors.forename: Dontscho
authors.initials: D
authors.lastname: Kerjaschki
authors.affiliation: Tohoku University Graduate School of Medicine, Department of Ophthalmology, Sendai, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Nakazawa
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kinoshita
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D003315:Cornea / Q000098:blood supply* / Q000187:drug effects / Q000473:pathology; D016510:Corneal Neovascularization / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D016039:Corneal Transplantation; D004195:Disease Models, Animal; D004729:Endothelium, Lymphatic / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D004797:Enzyme-Linked Immunosorbent Assay; D042583:Lymphangiogenesis / Q000187:drug effects*; D008264:Macrophages / Q000378:metabolism / Q000473:pathology; D008297:Male; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects; D000098853:Podoplanin
chemical_list: D008562:Membrane Glycoproteins; D020928:Mitogen-Activated Protein Kinases; D014409:Tumor Necrosis Factor-alpha; C490441:Pdpn protein, mouse; D000098853:Podoplanin
keywords: lymphangiogenesis; macrophage; podoplanin