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Edible mushroom (Agaricus bisporus) lectin modulates human retinal pigment epithelial cell behaviour in vitro
D Kent1, C Sheridan, H A Tomkinson
1Unit of Ophthalmology, Department of Medicine, University of Liverpool, Daulby Buildings, Liverpool L69 3GA, London, UK. dkent@liv.ac.uk
Abstract:
The retinal pigment epithelium (RPE) plays a major role in the development of proliferative vitreoretinopathy (PVR). In particular, RPE cells are implicated in generating the contraction forces seen. The present study was undertaken to investigate whether human RPE binds a lectin from the common edible mushroom, Agaricus bisporus, and to evaluate the effect of any binding on RPE-mediated matrix contraction in an in vitro model of PVR. Fluorescein isothiocyanate (FITC)-labelled Agaricus bisporus lectin (ABL) was used to study binding of lectin to normal retina, PVR scar tissue specimens and cultured human RPE. The effect of a 3-day exposure of ABL on human RPE-mediated contraction was evaluated using 2- and 3D RPE-populated collagen matrices. Effect of ABL on cell adhesion was measured using a collagen type I adhesion assay and determining the relative cellular attachment using absorbance readings. The normal RPE monolayer did not stain with FITC-ABL while PVR scar tissue stained intensely. Staining of in vitro RPE was characteristic but time-dependent. ABL caused a dose-dependent inhibition of RPE-mediated contraction of both 2D (one-way ANOVA, F = 7.94, p < 0.008) and 3D collagen matrices (one-way ANOVA, F = 164.955, p < 0.001). Pre-incubation of ABL with RPE in the 2D model caused a dramatic arrest of contraction (one-way ANOVA, F = 20.1, p < 0.001) that was due to a dose-dependent inhibition of adhesion (one-way ANOVA, F = 15.603, p < 0.001). Recovery of contraction was partially reversible on removal of ABL and was dependent on initial concentration of the lectin. ABL inhibits contraction and adhesion of human RPE cells in vitro without apparent cytotoxicity. It therefore deserves consideration as a potential therapeutic agent in the prevention and treatment of PVR and other non-ocular anomalous wound-healing processes.
Insights
Agaricus bisporus lectin (ABL) inhibits retinal pigment epithelium (RPE) cell contraction and adhesion in vitro. This lectin shows potential as a therapeutic agent for proliferative vitreoretinopathy (PVR) and other wound-healing disorders.
Area of Science:
- Ophthalmology
- Cell Biology
- Biochemistry
Background:
- Proliferative vitreoretinopathy (PVR) is a severe complication of retinal detachment, characterized by the contraction of retinal pigment epithelium (RPE) cells.
- RPE cells are key contributors to the fibrotic scarring and contraction that define PVR.
Purpose of the Study:
- To investigate if Agaricus bisporus lectin (ABL) binds to human RPE cells.
- To evaluate the effect of ABL on RPE-mediated matrix contraction in an in vitro model of PVR.
Main Methods:
- FITC-labeled ABL was used to assess lectin binding to normal retina, PVR scar tissue, and cultured human RPE.
- RPE-populated 2D and 3D collagen matrices were used to evaluate ABL's effect on RPE-mediated contraction and cell adhesion over 3 days.
Main Results:
- ABL intensely stained PVR scar tissue but not normal RPE; in vitro RPE staining was time-dependent.
- ABL significantly inhibited RPE-mediated contraction in both 2D and 3D collagen matrices in a dose-dependent manner.
- ABL dose-dependently inhibited RPE cell adhesion, leading to contraction arrest; partial recovery was observed upon ABL removal.
Conclusions:
- ABL inhibits human RPE cell contraction and adhesion in vitro without cytotoxicity.
- ABL demonstrates potential as a therapeutic agent for preventing and treating PVR and other non-ocular wound-healing processes.