Expression of activin A.

<p>Activin A expression was determined using anti-activin A antibodies (goat polyclonal; 1∶50; R&D Systems; Minneapolis, MN, USA). <b>A.</b> Case 17; infected endophthalmitis. Activin A was expressed in the corneal epithelial and the subepithelial space with inflammatory cells. <b>B.</b> Case 19; bacterial endophthalmitis. Activin A was expressed in the endothelium of newly formed blood vessels, which suggests the presence of vessel formation in the central cornea.</p

Distribution of dendritic cells (DCs).

<p>DCs were observed under a light microscope. Primary antibodies against CD1a (MTB1; mouse IgG1, κ; 1∶30 dilution; Novocastra; Newcastle-upon-Tyne, UK), DC-SIGN (CD209; H-200; rabbit polyclonal; 1∶400; Santa Cruz Biotechnology; Santa Cruz, CA, USA), langerin (CD207; 12D6; mouse IgG2b; 1∶100; Ylem S.R.L.; Rome, Italy), and CD83 (1H4b; mouse IgG1, κ; 1∶40; Novocastra) were used to observe DCs. <b>A.</b> Case 17; infected endophthalmitis. CD1a⁺ DCs were observed mainly in the epithelium and the subepithelial space of an inflamed cornea. <b>B.</b> Case 17; infected endophthalmitis. Langerin⁺ DCs were observed in the epithelium. <b>C.</b> Case 18; infected keratitis. DC-SIGN⁺ DCs were observed in the stroma. <b>D.</b> Case 22; corneal perforation, post PKP. CD83⁺ DCs were observed in the subepithelial space together with lymphocytic infiltration.</p

Increasing levels of cytokines in the supernatant of human corneal epithelial (HCE) cells.

<p><b>A.</b> Levels of activin A in the supernatants from group A (HCE cells), B (HCE cells stimulated with 5 µg/mL LPS), C (HCE cells co-cultured with 10⁵/well, and D (HCE cells co-cultured with imDCs and stimulated with LPS for 1 hour, 3 hours, and 6 hours). The levels of activin A increased in a time-dependent manner in supernatants from group B. After 6 hours, they were higher in groups B and D than in group C (<i>p</i><0.01, Uni-ANOVA with a Bonferroni correction). <b>B.</b> Levels of IL-6 in the supernatants of the groups described in <b>A.</b> The levels of IL-6 increased in a time-dependent manner in groups B, C, and D. The levels of IL-6 increased after 6 hours in supernatants from groups B, C, and D, but were undetectable in those from group A. There were no significant differences in the levels of IL-6 among the supernatants from groups B, C, and D after 6 hours. <b>C.</b> Levels of TNF-α in supernatants from the groups described in <b>A.</b> TNF-α was not detectable in those from group A. TNF-α levels increased significantly only in group D, and the increase was time-dependent (<i>p</i><0.01, Uni-ANOVA with a Bonferroni correction).</p

Numbers of dendritic cells (DCs) that migrated toward the supernatants of human corneal epithelial cells that were either stimulated or not stimulated with lipopolysaccharide (LPS).

A)<p>Mann-Whitney U test,</p><p>*<i>p</i><0.05.</p><p>The numbers of migratory dendritic cells in each well of the Boyden chamber are shown. LPS (−), supernatants from human corneal epithelial cells without LPS stimulation. LPS (+), supernatants from human corneal epithelial cells stimulated with LPS for 1.5 hours.</p><p>Numbers of dendritic cells (DCs) that migrated toward the supernatants of human corneal epithelial cells that were either stimulated or not stimulated with lipopolysaccharide (LPS).</p

Numbers of dendritic cells and vessels (per mm²) in the central area of the cornea.

A)<p>Mann-Whitney U test,</p><p>*<i>p</i><0.05.</p><p>There were statistically significant differences in the numbers of langerin⁺ DCs, DC-SIGN⁺ DCs, CD83⁺ DCs, and lymphatic vessels between infected and non-infected cases. Herpetic keratitis cases were excluded from the statistical analyses. This is because they were all in chronic states without neutrophil infiltration; therefore, infections could not be defined.</p><p>Numbers of dendritic cells and vessels (per mm²) in the central area of the cornea.</p