AsialoGM1-mediated IL-8 release by human corneal epithelial cells requires coexpression of TLR5.

PURPOSE: In this study, it was determined that human corneal epithelial cells (HCECs) express asialoganglioside ganliotetraosylceramide (asialoGM1) and toll-like receptor (TLR)-5, and their interaction induces interleukin (IL)-8 release through Ca(2+) transient activation and mitogen-activated protein kinase (MAPK) stimulation. METHODS: Expression of asialoGM1 and TLR5 was detected in SV40 HCECs by Western blot and flow cytometry analyses and their association by coimmunoprecipitation. Single-cell fluorescence imaging was used to measure intracellular free Ca(2+) transients in fura-2-loaded cells. The enzyme-linked immunosorbent assay (ELISA) was used to quantify IL-8 production in both cultured and primary HCECs. RESULTS: The HCECs expressed both asialoGM1 and TLR5 receptors. Ligation of asialoGM1 resulted in protein-protein interaction with TLR5, followed by transient increases in Ca(2+) influx through L-type voltage-dependent Ca(2+) channels. This led to P2Y receptor stimulation along with membrane depolarization, resulting from increases in ATP release into the medium. Intracellular Ca(2+) transients led to time-dependent extracellular signal-regulated kinase (ERK) MAPK pathway stimulation, followed by a 9.5-fold increase in IL-8 release. Similarly, in primary HCECs, asialoGM1 receptor stimulation resulted in an 8.1-fold increase. With a TLR5 neutralizing antibody, no asialoGM1-induced increases in IL-8 release occurred, and this response was not suppressed in the presence of a TLR2 neutralizing antibody. CONCLUSIONS: IL-8 release by HCECs is mediated through ligand-induced asialoGM1 protein-protein interactions with TLR5. This response is dependent on ATP efflux into the medium, followed by P2Y receptor stimulation. Such activation, in turn, results in increases in Ca(2+) influx through L-type voltage-dependent Ca(2+) channels, as well as stimulation of the ERK pathway.

Retrospective analysis of refractive errors in children with vision impairment.

PURPOSE: Emmetropization is the reduction in neonatal refractive errors that occurs after birth. Ocular disease may affect this process. We aimed to determine the relative frequency of ocular conditions causing vision impairment in the pediatric population and characterize the refractive anomalies present. We also compared the causes of vision impairment in children today to those between 1974 and 1981. METHODS: Causes of vision impairment and refractive data of 872 children attending a pediatric low-vision clinic from 1985 to 2002 were retrospectively collated. As a result of associated impairments, refractive data were not available for 59 children. An analysis was made of the causes of vision impairment, the distribution of refractive errors in children with vision impairment, and the average type of refractive error for the most commonly seen conditions. RESULTS: We found that cortical or cerebral vision impairment (CVI) was the most common condition causing vision impairment, accounting for 27.6% of cases. This was followed by albinism (10.6%), retinopathy of prematurity (ROP; 7.0%), optic atrophy (6.2%), and optic nerve hypoplasia (5.3%). Vision impairment was associated with ametropia; fewer than 25% of the children had refractive errors < or = +/-1 D. The refractive error frequency plots (for 0 to 2-, 6 to 8-, and 12 to 14-year age bands) had a Gaussian distribution indicating that the emmetropization process was abnormal. The mean spherical equivalent refractive error of the children (n = 813) was +0.78 +/- 6.00 D with 0.94 +/- 1.24 D of astigmatism and 0.92 +/- 2.15 D of anisometropia. Most conditions causing vision impairment such as albinism were associated with low amounts of hyperopia. Moderate myopia was observed in children with ROP. CONCLUSIONS: The relative frequency of ocular conditions causing vision impairment in children has changed since the 1970s. Children with vision impairment often have an associated ametropia suggesting that the emmetropization system is also impaired.