May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
A Genetic Polymorphism in the Human Toll-Like Receptor 3 Gene Protects Against dsRNA Induced Cell Death
Author Affiliations & Notes
  • W. Cho
    Ophthalmology and Visual Science, University of Kentucky, Lexington, Kentucky
  • M. E. Kleinman
    Ophthalmology and Visual Science, University of Kentucky, Lexington, Kentucky
  • D. Gibbs
    Ophthalmology and Visual Science, Moran Eye Center, University of Utah, Salt Lake City, Utah
    Program in Human Molecular Biology & Genetics, University of Utah, Eccles Institute of Human Genetics, Salt Lake City, Utah
  • D. R. Hinton
    Pathology and Ophthalmology, University of Southern California Keck School of medicine, Doheny Eye Institute, Los Angeles, California
  • K. Zhang
    Ophthalmology and Visual Science, Moran Eye Center, University of Utah, Salt Lake City, Utah
    Program in Human Molecular Biology & Genetics, University of Utah, Eccles Institute of Human Genetics, Salt Lake City, Utah
  • J. Ambati
    Ophthalmology and Visual Science, University of Kentucky, Lexington, Kentucky
  • Footnotes
    Commercial Relationships  W. Cho, None; M.E. Kleinman, None; D. Gibbs, None; D.R. Hinton, None; K. Zhang, None; J. Ambati, None.
  • Footnotes
    Support  NEI/NIH, Research to Prevent Blindness, Burroughs Wellcome Fund, American Health Assistance Foundation
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5156. doi:https://doi.org/
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      W. Cho, M. E. Kleinman, D. Gibbs, D. R. Hinton, K. Zhang, J. Ambati; A Genetic Polymorphism in the Human Toll-Like Receptor 3 Gene Protects Against dsRNA Induced Cell Death. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5156. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Toll-like receptor 3 (TLR3) is a double-stranded RNA (dsRNA) sensor that mediates the mammalian innate immune response against numerous viruses. Recently, a single nucleotide polymorphism (SNP) within the coding region of TLR3, termed L412F, was associated with a decreased risk of age-related macular degeneration, a disease hallmarked by retinal pigment epithelium (RPE) cell degeneration. There is preliminary evidence that this SNP suppresses dsRNA induced immune system activation and downstream host-cell death pathways; therefore, we sought to determine whether the L412F SNP affects RPE cell survival after exposure to dsRNA in vitro and in vivo.

Methods: : Primary RPE cells were genotyped for the TLR3 L412F SNP, and homozygous normal and heterozygous isolates were treated with various concentrations of the synthetic dsRNA, poly I:C, after sensitization with interferon α/β (1000U/mL). Annexin-V staining was analyzed by flow cytometry (FACS) at 24 hrs, and cell viability was assayed by BrdU ELISA at 48 hrs after poly I:C treatment. To evaluate the importance of TLR3 signaling in RPE cell death in vivo, we intravitreously injected poly I:C (2µg) or control poly dI:dC (2µg) into wild-type and TLR3 deficient mice. RPE cell survival and caspase-3 activation were quantified using FACS.

Results: : Human RPE cell isolates derived from carriers of the L412F SNP demonstrated increased cell viability and decreased apoptosis compared to non-carrier controls after exposure to the synthetic dsRNA, poly I:C. In vivo, we found that intravitreous administration of poly I:C in wild-type mice significantly decreased RPE cell numbers over a 72 hr time-course and increased RPE cell caspase-3 activation compared to poly dI:dC treated wild-type or TLR3 deficient mice.

Conclusions: : These data suggest that TLR3 signaling is critical in dsRNA mediated RPE cell death, and that patients with the L412F SNP may exhibit a decreased risk of AMD due to suppression of RPE cell death after exposure to viral dsRNA.

Keywords: age-related macular degeneration • apoptosis/cell death • mutations 
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