Figure 7
IL-6 neutralization blocks progression but does not induce regression of newly acquired lymphatic and blood vessels. Mice were infected and neutralizing antibody against IL-6 or isotype control antibody (1 μg/10 μl PBS) were injected subjconjunctivally at days 8, 10, and 12 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-IL-6 antibody-treated mice. Metamorph quantification of corneal area containing (b) LYVE-1+ lymphatic vessels, and (c) CD31+ blood vessels per 100X field of view. (d) Protein expression of pro-angiogenic factors at day 14 pi comparing isotypic control to anti-IL-6 antibody-treated mice. (e) Representative Z-stacked corneal images depicting lymphatic (green) and blood (red) vessels at day 21 pi comparing IgG to anti-IL-6-treated mice. Metamorph quantification of corneal area covered by (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-IL-6 antibody-treated mice. Data represent summary of the 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. *P<0.1, **P<0.01, ***P<0.001 (a–c,e–g) 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.
