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Impressum
Lymphatic capillaries in vascularized human corneas: ultrastructural and immunohistochemical evidence using VEGF receptor 3/flt-4, podoplanin and LYVE-1 as specific markers
1Cursiefen C., 1Schlötzer-Schrehardt U., 2Breiteneder-Geleff S., 4Alitalo K., 3Jackson D., 1Küchle M.
1Dept. of Ophthalmology, University of Erlangen, Germany; 2Dept. of Clinical Pathology, University of Vienna, Austria; 4Haartman Institute, Molecular/Cancer Biology Laboratory, University of Helsinki, Finland; 3MRC Human Immunology Unit, Institute of Molecular Medicine Oxford, England
Objective: To demonstrate immunohistochemical and ultrastructural
evidence for the presence of lymphatic capillaries in vascularized human
corneas.
Methods: Indirect immunohistochemistry was performed in vascularized
human corneas obtained by keratoplasty using specific markers of lymphatic
endothelium: a monoclonal antibody against VEGF receptor 3/flt-4 and polyclonal
antibodies against podoplanin and LYVE-1 (n=15; 4 µm paraffinembedded
sections). Electron microscopically, these corneas were evaluated for
presence of capillaries with the ultrastructural features of lymphatic
capillaries. Corneal neovascularization was secondary to acute and chronic
keratitis, transplant rejection, trauma and limbal insufficiency. Immunogoldlabeling
was performed for LYVE-1.
Results: Two types of capillaries were identified by immunohistochemistry
within the corneal stroma and subepithelially: erythrocyte-filled vessels
partly reacting with VEGF R3 but not with podoplanin and LYVE-1 and erythrocytefree
vessels reacting both with VEGF R3, LYVE-1 and podoplanin. Ultrastructurally,
two types of capillaries were detected, too: erythrocyte-filled vessels
with multilayered basement membrane, tight junctions and pericytes ("blood
capillaries") as well as erythrocyte-free vessels lined by flat endothelium
without tight junctions, without basement membrane and lacking pericytes
("lymphatic capillaries"). Lymphatic capillaries were less frequently
observed compared to blood capillaries and were always associated with
corneal inflammatory cells.
Conclusions: Immunohistochemical and ultrastructural observations
suggest lymphatic capillaries in vascularized human corneas. Corneal lymphangiogenesis
could partly explain the increased rate of transplant rejections occuring
in vascularized host corneas.
Support: BMBF (IZKF Erlangen, B13)