Figure 8
FGF-2 neutralization suppressed HSV-1-induced corneal neovascularization. Mice were inoculated with HSV-1, and neutralizing antibody against FGF-2 or isotype control antibody (5 μg per 10 μl PBS) were injected subjconjunctivally at days 8, 10, and 12 post infection (pi). (a) Representative Z-stacked corneal images depicting lymphatic (green) and blood (red) vessels at day 14 pi comparing isotype control antibody (IgG) to anti-FGF-2 antibody-treated mice. Metamorph quantification of corneal area containing (b) LYVE-1+ lymphatic vessels, and (c) CD31+ blood vessels per × 100 field of view. (d) Protein expression of pro-angiogenic factors at day 14 pi comparing isotypic control to anti-FGF-2 antibody-treated mice. (e) Representative Z-stacked corneal images depicting lymphatic (green) and blood (red) vessels at day 21 pi comparing isotypic control to anti-FGF-2 antibody-treated mice. Metamorph quantification of corneal area containing (f) LYVE-1+ lymphatic vessels, and (g) CD31+ blood vessels per × 100 field of view. (h) Protein expression of pro-angiogenic factors at day 21 pi comparing isotypic control to anti-FGF-2 antibody-treated mice. (i) Leukocyte content in the cornea at day 21 pi indicating (from left to right) neutrophils, macrophages, inflammatory monocytes, CD4+ T cells, CD8+ T cells, and NK cells. Data represent summary of mean±s.e.m. of two independent experiments (a–c,e–g) with n=10 corneas, three independent experiments (d,h) with n=6–12 mice, three independent experiments (h,i) with n=10–11 mice. **P<0.01, **P<0.001, ****P<0.0001 (a–c,e–g,i) as determined by Student’s t-test. *P<0.1, **P<0.01, ***P<0.001, ****P<0.0001 (d,h) as determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Bars=200 μm.
