Abstract
P 067
Inhibition of human retinal pigment epithelial cell attachment, spreading, and migration by the human Lectin Galectin-1
Claudia Priglinger1, Sabine André2, Thomas Kreutzer3, Anselm Kampik3, Hans-Joachim Gabius2, Siegfried Priglinger4
1Biomed-zet Life Science Laboratory, Linz, Augenklinik AKH Linz, Augenklinik der LMU München, München, 2Institut für physikalische Chemie, Veterinärmedizinische Fakultät, Ludwig-Maximilians-Universität, München, 3Augenklinik der Ludwig-Maximilians-Universität München, München, 4Augenklinik des AKH Linz, Linz, Österreich
Objective
Adhesion and migration of dislocated retinal pigment epithelial (RPE) cells is an initial step in pathogenesis of proliferative vitreoretinopathy (PVR). Lectins are glycoside-binding proteins which via lectin-carbohydrate interactions can influence a variety of cellular responses. With therapeutic perspective, the influence of the endogenous lectin, galectin-1, on attachment, spreading, and migration of human RPE cells was investigated.
Methods
Monolayer cultures of human RPE cells were treated with different concentrations of galectin-1 (0 - 250 µg/mL). Cell viability was tested by MTT assay. Galectin-1 binding to the RPE cells was investigated immunocytochemically. Attachment was assessed on extracellular matrix glycoproteins laminin or fibronectin, or on galectin-1, or the glycoprotein/lectin combinations coated on 96-well plates and subsequent MTT-testing. RPE migration in the absence or presence of galectin-1 on the respective substrates was tested using a modified Boyden chamber assay with PDGF-BB as chemoattractant. Cellular spreading was characterized by cytoplasmic halo formation of RPE cells after three hours in contact with the surface coating.
Results
Galectin-1 bound to the cell surface in a ß-galactoside-inhibitable manner. MTT assaying revealed no toxicity within control limits for the concentration range tested. When added to the medium, galectin-1 dose dependently inhibited RPE cell attachment, spreading and migration by more than 70%, irrespective of the substratum tested. When coated on the plastic surface, galectin-1 alone impaired spreading and migration of RPE cells, and reduced attachment to and migration on fibronectin by up to 80%.
Conclusions
Galectin-1, acting as a lectin on the cell surface, inhibits RPE cell attachment, migration, and spreading in vitro without apparent cytotoxicity. This activity of the endogenous effector deserves consideration as a potential therapeutic agent in the prevention and treatment of PVR.
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