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Akitomo Narimatsu, Takaaki Hattori, Naoto Koike, Kazuki Tajima, Hayate Nakagawa, Haruki Katahira, Teruumi Minezaki, Shigeto Kumakura, Tetsuya Matsumoto, Hiroshi Goto; Involvement of corneal lymphangiogenesis and macrophages in a murine bacterial keratitis model.. Invest. Ophthalmol. Vis. Sci. 2017;58(8):5767.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate corneal lymphangiogenesis and involvement of tissue-resident and monocyte-derived macrophages in a bacterial keratitis model in mice using Pseudomonas aeruginosa.
The mouse bacterial keratitis model was established using Pseudomonas aeruginosa strain PAO-1 and C57BL/6 mice. Strain PAO-1 (1×105 CFU/2.5ul) were applied in the infected group and PBS was applied in the control group after corneal epithelium was scratched. Lymphangiogenesis, angiogenesis, and macrophage infiltration were evaluated by immunostaining using whole-mount cornea at days 2, 7 and 14 post-inoculation. Anti-CD31 antibody, anti-LYVE-1 antibody, anti-CD11b antibody and anti-F4/80 antibody were used for immunostaining. Alteration of lymphangiogenesis by macrophage depletion was also evaluated intraperitoneal injection of clodronate-containing liposomes. Tissue-resident macrophages were depleted by sub-conjunctival injection and eye drop. Conversely, monocyte-derived macrophages were depleted by intraperitoneal injection.
Lymphangiogenesis and angiogenesis increased significantly in infected group at days 7 and 14 post-inoculation of PAO-1. A significant number of macrophages were observed around the lymphatic vessels in infected group. Lymphatic vessel formation was significantly reduced by monocyte-derived macrophage depletion, but not reduced by tissue-resident macrophage depletion in infected group.
These results suggest that the process of lymphangiogenesis in bacterial infection of the cornea occurs at the late stage of infection presumably induced by local infiltration of monocyte-derived macrophages.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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