Sub-lytic C5b-9 induces functional changes in retinal pigment epithelial cells consistent with age-related macular degeneration
| Autor: | Albrecht Lommatzsch, John Greenwood, Timothy R. Hughes, Jennifer A. E. Williams, Sarah E. Moss, Daniel Pauleikhoff, Katharina Lueck, Bryan Paul Morgan, Susanne Wasmuth |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Vascular Endothelial Growth Factor A
Pathology medicine.medical_specialty CD59 Complement Membrane Attack Complex Retinal Pigment Epithelium Immunofluorescence Cell Line chemistry.chemical_compound Macular Degeneration Laboratory Study medicine Humans Viability assay Interleukin 8 Vitronectin Complement Activation Chemokine CCL2 medicine.diagnostic_test biology Interleukin-6 Interleukin-8 Molecular biology Immunohistochemistry eye diseases Matrix Metalloproteinases Complement system Vascular endothelial growth factor Ophthalmology chemistry biology.protein sense organs Complement membrane attack complex |
| Popis: | PURPOSE: There is evidence for complement dysfunction in age-related macular degeneration (AMD). Complement activation leads to formation of the membrane attack complex (MAC), known to assemble on retinal pigment epithelial (RPE) cells. Therefore, the effect of sub-lytic MAC on RPE cells was examined with regard to pro-inflammatory or pro-angiogenic mediators relevant in AMD. METHODS: For sub-lytic MAC induction, RPE cells were incubated with an antiserum to complement regulatory protein CD59, followed by normal human serum (NHS) to induce 5% cell death, measured by a viability assay. MAC formation was evaluated by immunofluorescence and FACS analysis. Interleukin (IL)-6, -8, monocytic chemoattractant protein-1 (MCP-1), and vascular endothelial growth factor (VEGF) were quantified by enzyme-linked immunosorbent assay (ELISA). Intracellular MCP-1 was analysed by immunofluorescence, vitronectin by western blotting, and gelatinolytic matrix metalloproteinases (MMPs) by zymography. RESULTS: Incubation of RPE cells with the CD59 antiserum followed by 5% NHS induced sub-lytic amounts of MAC, verified by FACS and immunofluorescence. This treatment stimulated the cells to release IL-6, -8, MCP-1, and VEGF. MCP-1 staining, production of vitronectin, and gelatinolytic MMPs were also elevated in response to sub-lytic MAC. CONCLUSIONS: MAC assembly on RPE cells increases the IL-6, -8, and MCP-1 production. Therefore, sub-lytic MAC might have a significant role in generating a pro-inflammatory microenvironment, contributing to the development of AMD. Enhanced vitronectin might be a protective mechanism against MAC deposition. In addition, the increased expression of gelatinolytic MMPs and pro-angiogenic VEGF may be associated with neovascular processes and late AMD. |
| Databáze: | OpenAIRE |
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