Oxidative damage–induced inflammation initiates age-related macular degeneration


Oxidative damage and inflammation are postulated to be involved in age-related macular degeneration (AMD). However, the molecular signal(s) linking oxidation to inflammation in this late-onset disease is unknown. Here we describe AMD-like lesions in mice after immunization with mouse serum albumin adducted with carboxyethylpyrrole, a unique oxidation fragment of docosahexaenoic acid that has previously been found adducting proteins in drusen from AMD donor eye tissues1 and in plasma samples2 from individuals with AMD. Immunized mice develop antibodies to this hapten, fix complement component-3 in Bruch's membrane, accumulate drusen below the retinal pigment epithelium during aging, and develop lesions in the retinal pigment epithelium mimicking geographic atrophy, the blinding end-stage condition characteristic of the dry form of AMD. We hypothesize that these mice are sensitized to the generation of carboxyethylpyrrole adducts in the outer retina, where docosahexaenoic acid is abundant and conditions for oxidative damage are permissive. This new model provides a platform for dissecting the molecular pathology of oxidative damage in the outer retina and the immune response contributing to AMD.

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Figure 1: CEP antigen and CEP-specific antibody titers in immunized mice.
Figure 2: Relationship between pathology and CEP-specific antibody titer in the short-term immunization mice.
Figure 3: C3d localization in the outer eye wall of the short-term immunization mice.
Figure 4: Light and electron microscopy of the RPE–Bruch's membrane interface with quantitative comparisons.


  1. 1

    Crabb, J.W. et al. Drusen proteome analysis: an approach to the etiology of age-related macular degeneration. Proc. Natl. Acad. Sci. USA 99, 14682–14687 (2002).

    CAS  Article  Google Scholar 

  2. 2

    Gu, X. et al. Carboxyethylpyrrole protein adducts and autoantibodies, biomarkers for age-related macular degeneration. J. Biol. Chem. 278, 42027–42035 (2003).

    CAS  Article  Google Scholar 

  3. 3

    Javitt, J.C., Zhou, Z., Maguire, M.G., Fine, S.L. & Willke, R.J. Incidence of exudative age-related macular degeneration among elderly Americans. Ophthalmology 110, 1534–1539 (2003).

    Article  Google Scholar 

  4. 4

    Augood, C.A. et al. Prevalence of age-related maculopathy in older Europeans: the European Eye Study (EUREYE). Arch. Ophthalmol. 124, 529–535 (2006).

    Article  Google Scholar 

  5. 5

    Hageman, G.S. et al. An integrated hypothesis that considers drusen as biomarkers of immune-mediated processes at the RPE-Bruch's membrane interface in aging and age-related macular degeneration. Prog. Retin. Eye Res. 20, 705–732 (2001).

    CAS  Article  Google Scholar 

  6. 6

    Anderson, D.H., Mullins, R.F., Hageman, G.S. & Johnson, L.V. A role for local inflammation in the formation of drusen in the aging eye. Am. J. Ophthalmol. 134, 411–431 (2002).

    CAS  Article  Google Scholar 

  7. 7

    Johnson, L.V., Ozaki, S., Staples, M.K., Erickson, P.A. & Anderson, D.H. A potential role for immune complex pathogenesis in drusen formation. Exp. Eye Res. 70, 441–449 (2000).

    CAS  Article  Google Scholar 

  8. 8

    Edwards, A.O. et al. Complement factor H polymorphism and age-related macular degeneration. Science 308, 421–424 (2005).

    CAS  Article  Google Scholar 

  9. 9

    Hageman, G.S. et al. A common haplotype in the complement regulatory gene factor H (HF1/CFH) predisposes individuals to age-related macular degeneration. Proc. Natl. Acad. Sci. USA 102, 7227–7232 (2005).

    CAS  Article  Google Scholar 

  10. 10

    Haines, J.L. et al. Complement factor H variant increases the risk of age-related macular degeneration. Science 308, 419–421 (2005).

    CAS  Article  Google Scholar 

  11. 11

    Klein, R.J. et al. Complement factor H polymorphism in age-related macular degeneration. Science 308, 385–389 (2005).

    CAS  Article  Google Scholar 

  12. 12

    Gold, B. et al. Variation in factor B (BF) and complement component 2 (C2) genes is associated with age-related macular degeneration. Nat. Genet. 38, 458–462 (2006).

    CAS  Article  Google Scholar 

  13. 13

    Yates, J.R. et al. Complement C3 variant and the risk of age-related macular degeneration. N. Engl. J. Med. 357, 553–561 (2007).

    CAS  Article  Google Scholar 

  14. 14

    Anderson, R.E., Lissandrello, P.M., Maude, M.B. & Matthes, M.T. Lipids of bovine retinal pigment epithelium. Exp. Eye Res. 23, 149–157 (1976).

    CAS  Article  Google Scholar 

  15. 15

    Anderson, R.E. Lipids of the ocular tissues. IV. A comparison of the phospholipids from the retina of six mammalian species. Exp. Eye Res. 10, 339–344 (1970).

    CAS  Article  Google Scholar 

  16. 16

    Fliesler, S.J. & Anderson, R.E. Chemistry and metabolism of lipids in the vertebrate retina. Prog. Lipid Res. 22, 79–131 (1983).

    CAS  Article  Google Scholar 

  17. 17

    Seddon, J.M., Willett, W.C., Speizer, F.E. & Hankinson, S.E. A prospective study of cigarette smoking and age-related macular degeneration in women. J. Am. Med. Assoc. 276, 1141–1146 (1996).

    CAS  Article  Google Scholar 

  18. 18

    Klein, R., Klein, B.E. & Cruickshanks, K.J. The prevalence of age-related maculopathy by geographic region and ethnicity. Prog. Retin. Eye Res. 18, 371–389 (1999).

    CAS  Article  Google Scholar 

  19. 19

    Snow, K.K. & Seddon, J.M. Do age-related macular degeneration and cardiovascular disease share common antecedents? Ophthalmic Epidemiol 6, 125–143 (1999).

  20. 20

    Christen, W.G., Glynn, R.J., Manson, J.E., Ajani, U.A. & Buring, J.E. A prospective study of cigarette smoking and risk of age-related macular degeneration in men. J. Am. Med. Assoc. 276, 1147–1151 (1996).

    CAS  Article  Google Scholar 

  21. 21

    Solberg, Y., Rosner, M. & Belkin, M. The association between cigarette smoking and ocular diseases. Surv. Ophthalmol. 42, 535–547 (1998).

    CAS  Article  Google Scholar 

  22. 22

    Peakman, M., Senaldi, G. & Vergan, D. Review: assessment of complement activation in clinical immunology laboratories: time for reappraisal? J. Clin. Pathol. 42, 1018–1025 (1989).

    CAS  Article  Google Scholar 

  23. 23

    Ambati, J. et al. An animal model of age-related macular degeneration in senescent Ccl-2– or Ccr-2–deficient mice. Nat. Med. 9, 1390–1397 (2003).

    CAS  Article  Google Scholar 

  24. 24

    Gottsch, J.D., Bynoe, L.A., Harlan, J.B., Rencs, E.V. & Green, W.R. Light-induced deposits in Bruch's membrane of protoporphyric mice. Arch. Ophthalmol. 111, 126–129 (1993).

    CAS  Article  Google Scholar 

  25. 25

    Cousins, S.W. et al. The role of aging, high-fat diet and blue light exposure in an experimental mouse model for basal laminar deposit formation. Exp. Eye Res. 75, 543–553 (2002).

    CAS  Article  Google Scholar 

  26. 26

    Malek, G. et al. Apolipoprotein E allele–dependent pathogenesis: a model for age-related retinal degeneration. Proc. Natl. Acad. Sci. USA 102, 11900–11905 (2005).

    CAS  Article  Google Scholar 

  27. 27

    Mombaerts, P. et al. RAG-1–deficient mice have no mature B and T lymphocytes. Cell 68, 869–877 (1992).

    CAS  Article  Google Scholar 

  28. 28

    Gu, X., Sun, M., Hazen, S., Crabb, J.W. & Salomon, R.G. Oxidatively truncated docosahexaenoate phosphoplipids: total synthesis, generation and peptide adduction chemistry. J. Org. Chem. 68, 3749–3761 (2003).

    CAS  Article  Google Scholar 

  29. 29

    Percopo, C.M., Hooks, J.J., Shinohara, T., Caspi, R. & Detrick, B. Cytokine-mediated activation of a neuronal retinal resident cell provokes antigen presentation. J. Immunol. 145, 4101–4107 (1990).

    CAS  PubMed  Google Scholar 

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J.G.H. and V.L.P. designed and initiated the experiments. L.L. and R.G.S. prepared the CEP-MSA and CEP-BSA. R.L.U., V.L.P., K.G.S. and X.Y. immunized the mice. K.G.S. and X.Y. performed the protein chemistry and ELISA assays, as well as managing the day-to-day maintenance of the mice. M.E.R. and J.G.H. performed all of the histological and electron microscopic analysis. V.L.B. performed the confocal microscopy. J.G.H. analyzed all of the data and wrote the manuscript. R.G.S., R.L.U. and V.L.P. made critical comments and suggestions for revisions of the manuscript in response to the reviewers.

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Supported by the State of Ohio Biomedical Research and Technology Transfer Program, a Research Center Grant from the Foundation Fighting Blindness, and a Challenge Grant from Research to Prevent Blindness. The project was also supported by grants R56EY10240, R01EY014240, R24EY015638 (J.G.H.), R21EY017153 (V.L.B.), R01GM21249 (R.G.S.) and K08EY014912 (V.L.P.) from the US National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the US National Eye Institute of the National Institutes of Health. We thank J.W. Crabb for valuable discussions, B. Anand-Apte for critical comments on the manuscript, N.S. Peachey for help with the statistical comparisons, R.L. Fairchild for providing the Rag-deficient mice, Y. Li for help with the histology, K. Sayanagi and T. Yakamoto for the fundus photography and A. Vasanji for developing the image-analysis algorithm used to define the areas of sub-RPE deposits.

Corresponding authors

Correspondence to Joe G Hollyfield or Victor L Perez.

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Competing interests

The mouse model for age-related macular degeneration described in this study is protected for commercialization by the Cleveland Clinic. J.G.H., R.G.S. and V.L.P. are the inventors.

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Supplementary Figs. 1–7 and Supplementary Table 1 (PDF 1014 kb)

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Hollyfield, J., Bonilha, V., Rayborn, M. et al. Oxidative damage–induced inflammation initiates age-related macular degeneration. Nat Med 14, 194–198 (2008). https://doi.org/10.1038/nm1709

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