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Hem Raj Gurung, Daniel J Carr; Neutralization of Fibroblast Growth Factor-2 (FGF-2) Suppresses HSV-1-induced Corneal Lymphangiogenesis. Invest. Ophthalmol. Vis. Sci. 2016;57(12):329.
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© ARVO (1962-2015); The Authors (2016-present)
The genesis of lymphatic vessels (lymphangiogenesis) is induced in murine cornea following herpes simplex virus type 1 (HSV-1) infection. Lymphangiogenesis progresses and is maintained well past the resolution of infection by unknown means. Previous studies by our group suggest infiltrating leukocytes do not play a role. The present study was undertaken to identify the contribution of pro-angiogenic factors that promote and sustain lymphangiogenesis past virus clearance.
C57BL6/J mice were infected with 500-1,000 plaque forming units (PFU) of HSV-1 per eye. Ten mg/kg dexamethasone (DEX) or vehicle was administered by intraperitoneal injection at day 10 pi. FGF-2 and IL-6 were neutralized in vivo by subconjunctival injections of anti-FGF-2 or anti-IL-6 monoclonal antibodies respectively on days 8, 10, and 12 pi. Mice were euthanized at day 14 and day 21 pi, and the corneas were removed and processed. Corneal lymphatic vessels (LYVE-1+) and blood vessels (CD31+) were visualized by confocal microscopy following immunohistochemical staining. Pro-angiogenic protein (angiopoietin-2, FGF-2, hepatocyte growth factor, interleukin [IL]-1, IL-6, matrix metalloproteinase 9, tumor necrosis factor [TNF]-α, vascular endothelial growth factor [VEGF]-A, and VEGF-C) levels were determined by bioplex multiplex array or ELISA at times pi. Data were analyzed by ANOVA and Tukey’s multiple comparisons test.
A single bolus of DEX at day 10 pi resulted in a significant reduction of blood vessel genesis (hemangiogenesis) at day 14 pi and blood and lymphatic vessel development into the cornea at day 21 pi compared to control-treated mice. Among factors identified in the cornea at times pi impacted by DEX, we focused on FGF-2 and IL-6. Whereas IL-6 neutralization had a modest impact on hemangiogenesis (day 14-21 pi) and lymphangiogenesis (day 21 pi) in a time-dependent fashion, neutralization of FGF-2 had a more pronounced effect in suppressing neovascularization (blood and lymphatic vessels) in total in a time-dependent manner.
A single bolus of DEX at the time of HSV-1 clearance completely reverses the incidence of angiogenesis in the cornea of mice. This process was recapitulated by antibody-mediated neutralization of FGF- 2.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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