Increased Epithelial Expression of CTGF and S100A7 with Elevated Subepithelial Expression of IL-1β in Trachomatous Trichiasis

Abstract

Purpose

To characterize the histological appearance and expression of pro-inflammatory mediators, growth factors, matrix metalloproteinases and biomarkers of epithelial-mesenchymal transition (EMT) in healthy control and trachomatous trichiasis (TT) conjunctival tissue.

Methods

Conjunctival biopsies were taken from 20 individuals with TT and from 16 individuals with healthy conjunctiva, which served as controls. Study participants were of varying ethnicity and were living in a trachoma-endemic region of northern Tanzania. Formalin-fixed paraffin-embedded tissue sections were stained using hematoxylin and eosin or by immunohistochemistry using antibodies against: IL-1β, IL-6, IL-17A, IL-22, CXCL5, S100A7, cleaved caspase 1 (CC1), PDGF, CTGF, TGFβ2, MMP7, MMP9, E-cadherin, vimentin, and αSMA.

Results

Tissue from TT cases had a greater inflammatory cell infiltrate relative to controls and greater disruption of collagen structure. CTGF and S100A7 were more highly expressed in the epithelium and IL-1β was more highly expressed in the substantia propria of TT cases relative to controls. Latent TGFβ2 was slightly more abundant in the substantia propria of control tissue. No differences were detected between TT cases and controls in the degree of epithelial atrophy, the number of myofibroblasts or expression of EMT biomarkers.

Conclusions

These data indicate that the innate immune system is active in the immunopathology of trachoma, even in the absence of clinical inflammation. CTGF might provide a direct link between inflammation and fibrosis and could be a suitable target for therapeutic treatment to halt the progression of trachomatous scarring.

Author Summary

Progressive scarring of the conjunctiva in individuals with trachoma causes the eyelids to contract, drawing the eyelashes inwards (trichiasis) so that they scratch the cornea, causing pain and eventually blindness. Disease is initiated in childhood by repeated conjunctival infection with Chlamydia trachomatis (Ct), however, infection is not commonly found in adults, yet chronic inflammation and fibrosis progress throughout the lives of many individuals. A better understanding of the mechanisms driving inflammation and fibrosis are required in order to develop treatments to halt disease progression. The tissue expression and localization of a number of pro-inflammatory cytokines, growth and matrix factors were investigated in eyelid tissue from 20 individuals with trichiasis and from 16 control individuals. By staining tissue sections with dyes and specific antibodies, pro-inflammatory signaling molecules IL-1β and S100A7 and pro-fibrotic growth factor CTGF were found to be more highly expressed in individuals with trichiasis. CTGF and S100A7 were highly expressed in the epithelium; the outermost layer of the conjunctiva, whereas IL-1β was more highly expressed deeper in the tissue, where scarring occurs. Numerous inflammatory cells were found in the tissue of trichiasis patients even in the absence of clinically apparent inflammation. Future research should seek to describe a causative mechanism linking these factors.