title: All-trans-retinal sensitizes human RPE cells to alternative complement pathway-induced cell death.
abstract: OBJECTIVE
  Retinal pigment epithelial (RPE) cell death occurs early in the pathogenesis of age-related macular degeneration (AMD) and Stargardt's disease. Emerging evidence suggests that all-trans-retinal (atRal) and alternative complement pathway (AP) activation contribute to RPE cell death in both of these retinal disorders. The aim of this study was to investigate the combined effect of atRal and AP activation on RPE cell viability.
  
  
  METHODS
  RPE cells were treated with atRal and then incubated with a complement-fixing antibody followed by stimulation with C1q-depleted serum to activate AP. Cell viability was assessed by tetrazolium salt and lactate dehydrogenase release assays. Changes in cell surface CD46 and CD59 expression were assessed by flow cytometry. Cells were pretreated with the antioxidant resveratrol, and C1q-depleted serum was incubated with an anti-C5 antibody prior to initiating AP attack to determine the protective effects of antioxidant therapy and complement inhibition, respectively.
  
  
  RESULTS
  Both atRal and AP activation independently caused RPE cell death. When AP attack was initiated following atRal treatment, a synergistic increase in cell death was observed. Following 24-hour atRal treatment, CD46 and CD59 expression decreased, corresponding temporally to increased susceptibility to AP attack. Resveratrol and the anti-C5 antibody both protected against AP-induced cell death following atRal exposure and were most effective when used in combination.
  
  
  CONCLUSIONS
  atRal sensitizes RPE cells to AP attack, which may be mediated in part by atRal-induced downregulation of CD46 and CD59. Despite increased susceptibility to AP attack following exposure to atRal, resveratrol and anti-C5 antibody effectively prevent AP-mediated cell death.
authors.affiliation: Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Jacob E
authors.initials: JE
authors.lastname: Berchuck
authors.forename: Ping
authors.initials: P
authors.lastname: Yang
authors.forename: Brett A
authors.initials: BA
authors.lastname: Toimil
authors.forename: Zhe
authors.initials: Z
authors.lastname: Ma
authors.forename: Peter
authors.initials: P
authors.lastname: Baciu
authors.forename: Glenn J
authors.initials: GJ
authors.lastname: Jaffe
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects / Q000276:immunology; D002478:Cells, Cultured; D003170:Complement Pathway, Alternative / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D008268:Macular Degeneration / Q000188:drug therapy / Q000276:immunology / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D000077185:Resveratrol; D055213:Retinal Pigment Epithelium / Q000187:drug effects / Q000276:immunology / Q000473:pathology*; D012264:Ribonucleotide Reductases / Q000037:antagonists & inhibitors; D013267:Stilbenes / Q000494:pharmacology
chemical_list: D020533:Angiogenesis Inhibitors; D013267:Stilbenes; D012264:Ribonucleotide Reductases; D000077185:Resveratrol